Dr, Hokkaido University

糖質加リン酸分解酵素

糖質異性化

マンノース含有オリゴ糖

オリゴ糖

α-グルコシグーゼ

酵素利用学

加水分解酵素

α-amylase

酵素反応機構

糖転移反応

酵素化学

イソマルトオリゴ糖

糖質関連酵素

多糖合成酵素

応用生物化学

Applied Biochemistry (6103)

Life Science, Applied biochemistry

Bachelor's degree program, School of Agriculture

Master's degree program, Graduate School of Agriculture

Doctoral (PhD) degree program, Graduate School of Agriculture

Sep. 2019, 日本応用糖質科学会, 学会賞
各種糖質加水分解酵素・加リン酸分解酵素・異性化酵素の機能と応用に関する研究
森 春英

2007, Best Poster Award

Sep. 2005, 日本応用糖質科学会, 奨励賞
植物α－アミラーゼの機能と構造に関する研究
森 春英

2004, ベストポスター賞

2003, B.B.B.論文賞
Japan

2003, Biosci Biotechnol Biochem Paper Award(JSBBA, 2003)

Structure and Function of Honeybee α-Amylase of Glycoside Hydrolase Family 13 Subfamily 15
Wataru Saburi; Yushi Takahashi; Shiho Takei; Toyoyuki Ose; Haruhide Mori
Molecules, 27 Jul. 2026
Scientific journal

Functional and Structural Diversity of Glycoside Hydrolase Family 15 Enzymes
Takayoshi Tagami; Wataru Saburi; Masayuki Okuyama; Haruhide Mori
Bulletin of Applied Glycoscience, 16, 2, 48, 56, The Japanese Society of Applied Glycoscience, 20 May 2026, [Peer-reviewed]
Japanese, Scientific journal

Insights into the recognition of cyclic α-(1→6)-glucan by a solute-binding protein of an ABC transporter from Tepidibacillus decaturensis
Shiho Takei; Wataru Saburi; Min Yao; Haruhide Mori; Toyoyuki Ose
Journal of Biological Chemistry, May 2026
Scientific journal

Mechanism for synthesis of isomaltooligosaccharides from maltooligosaccharides by GH15 α‐glucan 4(6)‐α‐glucosyltransferase
Tianyi Qin; Wataru Saburi; Momo Sawada Otsubo; Haruki Oshita; Kenta Kanai; Tomoya Ota; Birte Svensson; Haruhide Mori
The FEBS Journal, May 2026
Scientific journal

Functional Analysis of Amino Acid Residues Responsible for Substrate Specificity of GH13_17 α‑Glucosidase from Aedes aegypti Saliva (AaMalI).
Waraporn Auiewiriyanukul; Wataru Saburi; Haruhide Mori; Dumrongkiet Arthan; Sorachat Tharamak
ACS omega, 11, 10, 15795, 15808, 17 Mar. 2026, [International Magazine]
English, Scientific journal, The α-glucosidase (AaMalI) in Aedes aegypti saliva belongs to glycoside hydrolase family 13, subfamily 17 (GH13_17) and plays a crucial role in the digestion of sucrose, which is the main sugar involved in insect metabolism. The amino-acid residues in the conserved region II have been reported as the key residues for sucrose specificity in GH13_17. Using mutagenesis, this study expressed and purified recombinant AaMalI and determined the molecular mechanism related to substrate specificity. The optimal activity was at pH 6.3 and 40 °C. AaMalI had a trisaccharide specificity similar to GH13 α-glucosidases and preference for sucrose over maltose. The single mutation of Y223H and the double mutation of P222N/Y223H altered the substrate preference from sucrose to maltose. Structural analysis of the AaMalI model obtained by superimposition with the maltose-bound complex suggested that Tyr292 stabilizes the d-glucosyl moiety at subsite +1, whereas His223 indirectly contributes to maltose hydrolysis. These findings provide structural insights into AaMalI substrate specificity and support its potential as a target for vector mosquito control.

AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana.
Katsunari Oki; Akane Enoki; Yoshitaka Yokota; Taiki Kurihara; Takafumi Shimizu; Wataru Saburi; Takayuki Tohge; Haruhide Mori; Guido Van den Ackerveken; Naoki Kitaoka; Hideyuki Matsuura
Chembiochem : a European journal of chemical biology, e2500151, 11 Sep. 2025, [International Magazine]
English, Scientific journal, Jasmonates are plant hormones that regulate plant defense and development. 7-iso-Jasmonoyl-l-isoleucine (JA-Ile) is a representative active jasmonate which is biosynthesized from 7-iso-jasmonic acid (JA) by the jasmonoyl-amido synthases JASMONATE RESISTANT 1 (JAR1) and AtGH3.10 in Arabidopsis thaliana. 12-Hydroxy-7-iso-jasmonoyl-l-isoleucine (12-OH-JA-Ile) is another active jasmonate, and 12-hydroxylation of JA-Ile is considered the major biosynthetic pathway toward 12-OH-JA-Ile. Previous report elucidated that recombinant JAR1 showed a weak activity against 12-hydroxy-7-iso-jasmonic acid (12-OH-JA). However, the direct conversion from 12-OH-JA to 12-OH-JA-Ile in planta and the enzyme activity of AtG3.10 against 12-OH-JA have never been reported. Herein, a feeding experiment with deuterated 12-OH-JA confirms the direct conversion of 12-OH-JA to 12-OH-JA-Ile in wild-type Arabidopsis plants. The conversion from 12-OH-JA to 12-OH-JA-Ile is not observed in jar1 gh3.10 double mutant, suggesting that 12-OH-JA is converted to 12-OH-JA-Ile by JAR1 and AtGH3.10. Notably, enzyme assays show that the catalytic efficiency with 12-OH-JA for AtGH3.10 is higher than those with JA for AtGH3.10 and with 12-OH-JA for JAR1. Comparative analysis of JAR1 and AtGH3.10 structures and site-directed mutation analysis reveals that Ser120 in AtGH3.10 is the key amino acid residue responsible for its high catalytic efficiency against 12-OH-JA.

Structural basis of transglucosylation in dextran dextrinase, a homolog of anomer-inverting GH15 glucoside hydrolases
Takayoshi Tagami; Wataru Saburi; Juri Sadahiro; Yuya Kumagai; Weeranuch Lang; Naohiro Matsugaki; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
Journal of Biological Chemistry, 301, 9, 110541, 110541, Elsevier BV, Sep. 2025, [Peer-reviewed], [Corresponding author]
English, Scientific journal

Biochemical and structural analysis of the mechanism for the catalysis and specificity of cellobiose 2-epimerase from Rhodothermus marinus
Wataru Saburi; Hirohiko Muto-Fukiya; Nongluck Jaito; Koji Kato; Jian Yu; Min Yao; Haruhide Mori
Bioscience, Biotechnology, and Biochemistry, 89, 7, 973, 984, Oxford University Press (OUP), 22 Mar. 2025, [Last author, Corresponding author]
Scientific journal, Abstract

Cellobiose 2-epimerase (CE) catalyzes C-2 epimerization of reducing end d-glucose/d-mannose residue of β-(1→4)-disaccharides, and also slightly catalyzes aldose-ketose conversion. In this study, we investigated the structure-function relationship of Rhodothermus marinus CE (RmCEs). In 2H2O, 2H replaced the 2-H of the reducing end sugar residue, suggesting a proton abstraction-addition mechanism via the cis-enediolate intermediate. The structure of the RmCE-mannobiitol complex showed that His259 was suitable for abstracting 2-H from d-mannose residue, whereas His390 was suitable for the d-glucose residue. H259A and H390A mutations abolished activity for Galβ1-4Man and Galβ1-4Glc formation from Galβ1-4Fru, respectively, and these mutants catalyzed both epimerization and isomerization to Galβ1-4Glc and Galβ1-4Man, respectively. Ala substitution of the residues interacting with the 2-O of the reducing end sugar residue significantly reduced the velocity for epimerization, but not for isomerization. Trp385, stacked onto the non-reducing-end sugar residues of disaccharides, was shown to be important for disaccharide specificity.

Molecular mechanism for the substrate specificity of Arthrobacter globiformis M6 α-glucosidase CmmB, belonging to glycoside hydrolase family 13 subfamily 30
Wataru Saburi; Takayoshi Tagami; Takuya Usui; Jian Yu; Toyoyuki Ose; Min Yao; Haruhide Mori
Food Bioscience, 61, 104516, 104516, Elsevier BV, Oct. 2024, [Peer-reviewed], [Last author]
Scientific journal

Comparisons of the amylolytic enzymes and malt starch hydrolysates of two barley cultivars, Hokudai 1 (the first cultivar developed in Japan) and Kitanohoshi (currently used cultivar for beer production)
Wataru Saburi; Haruhide Mori
Bioscience, Biotechnology, and Biochemistry, 88, 10, 1180, 1187, Oxford University Press (OUP), 11 Jul. 2024, [Peer-reviewed], [Last author]
English, Scientific journal, Abstract

Starch degradation in malted barley produces yeast-fermentable sugars. In this study, we compared the amylolytic enzymes and composition of the malt starch hydrolysates of two barley cultivars, Hokudai 1 (the first cultivar established in Japan) and Kitanohoshi (the currently used cultivar for beer production). Hokudai 1 malt contained lower activity of amylolytic enzymes than Kitanohoshi malt, although these cultivars contained α-amylase AMY2 and β-amylase Bmy1 as the predominant enzymes. Malt starch hydrolysate of Hokudai 1 contained more limit dextrin and less yeast-fermentable sugars than that of Kitanohoshi. In mixed malt saccharification, a high Hokudai 1 malt ratio increased the limit dextrin levels and decreased the maltotriose and maltose levels. Even though Kitanohoshi malt contained more amylolytic enzymes than Hokudai 1 malt, addition of Kitanohoshi extract containing the amylolytic enzymes did not enhance malt starch degradation of Hokudai 1. Hokudai 1 malt starch was less degradable than Kitanohoshi malt starch.

Tunable structure of chimeric isomaltomegalosaccharides with double α-(1 → 4)-glucosyl chains enhances the solubility of water-insoluble bioactive compounds
Weeranuch Lang; Takayoshi Tagami; Yuya Kumagai; Seiya Tanaka; Hye-Jin Kang; Masayuki Okuyama; Wataru Saburi; Haruhide Mori; Tohru Hira; Chaehun Lee; Takuya Isono; Toshifumi Satoh; Hiroshi Hara; Takayuki Kurokawa; Nobuo Sakairi; Yoshiaki Yuguchi; Atsuo Kimura
Carbohydrate Polymers, 319, 121185, 121185, Elsevier BV, Nov. 2023, [Peer-reviewed]
Scientific journal

Molecular mechanism for endo-type action of glycoside hydrolase family 55 endo-β-1,3-glucanase on β1-3/1-6-glucan.
Tomoya Ota; Wataru Saburi; Takayoshi Tagami; Jian Yu; Shiro Komba; Linda Elizabeth Jewell; Tom Hsiang; Ryozo Imai; Min Yao; Haruhide Mori
The Journal of biological chemistry, 105294, 105294, 27 Sep. 2023, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, The glycoside hydrolase family 55 (GH55) includes inverting exo-β-1,3-glucosidases and endo-β-1,3-glucanases, acting on laminarin, which is a β1-3/1-6-glucan consisting of a β1-3/1-6-linked main chain and β1-6-linked branches. Despite their different modes of action toward laminarin, endo-β-1,3-glucanases share with exo-β-1,3-glucosidases conserved residues that form the dead-end structure of subsite -1. Here, we investigated the mechanism of endo-type action on laminarin by GH55 endo-β-1,3-glucanase MnLam55A, identified from Microdochium nivale. MnLam55A, like other endo-β-1,3-glucanases, degraded internal β-d-glucosidic linkages of laminarin, producing more reducing sugars than the sum of d-glucose and gentiooligosaccharides detected. β1-3-Glucans lacking β1-6-linkages in the main chain were not hydrolyzed. NMR analysis of the initial degradation of laminarin revealed that MnLam55A preferentially cleaved the non-reducing terminal β1-3-linkage of the laminarioligosaccharide moiety at the reducing end side of the main chain β1-6-linkage. MnLam55A liberates d-glucose from laminaritriose and longer laminarioligosaccharides, but kcat/Km values to laminarioligosaccharides (≤4.21 s-1mM-1) were much lower than to laminarin (5,920 s-1mM-1). These results indicate that β-glucan binding to the minus subsites of MnLam55A, including exclusive binding of the gentiobiosyl moiety to subsites -1 and -2, is required for high hydrolytic activity. A crystal structure of MnLam55A, determined at 2.4 Å resolution, showed that MnLam55A adopts an overall structure and catalytic site similar to those of exo-β-1,3-glucosidases. However, MnLam55A possesses an extended substrate-binding cleft that is expected to form the minus subsites. Sequence comparison suggested that other endo-type enzymes share the extended cleft structure. The specific hydrolysis of internal linkages in laminarin is presumably common to GH55 endo-β-1,3-glucanases.

Hydrolysis-transglycosylation of sucrose and production of β-(2→1)-fructan by inulosucrase from Neobacillus drentensis 57N.
Yusuke Kido; Wataru Saburi; Taizo Nagura; Haruhide Mori
Bioscience, biotechnology, and biochemistry, 87, 10, 1169, 1182, 21 Sep. 2023, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Inulin, β-(2→1)-fructan, is a beneficial polysaccharide used as a functional food ingredient. Microbial inulosucrases (ISs), catalyzing β-(2→1)-transfructosylation, produce β-(2→1)-fructan from sucrose. In this study, we identified a new IS (NdIS) from the soil isolate, Neobacillus drentensis 57N. Sequence analysis revealed that, like other Bacillaceae ISs, NdIS consists of a glycoside hydrolase family 68 domain and shares most of the 1-kestose-binding residues of the archaeal IS, InuHj. Native and recombinant NdIS were characterized. NdIS is a homotetramer. It does not require calcium for activity. High performance liquid chromatography and 13C-nuclear magnetic resonance indicated that NdIS catalyzed the hydrolysis and β-(2→1)-transfructosylation of sucrose to synthesize β-(2→1)-fructan with chain lengths of 42 or more residues. The rate dependence on sucrose concentration followed hydrolysis-transglycosylation kinetics, and a 50% transglycosylation ratio was obtained at 344 m m sucrose. These results suggest that transfructosylation from sucrose to β-(2→1)-fructan occurs predominantly to elongate the fructan chain because sucrose is an unfavorable acceptor.

Chemical synthesis of oligosaccharide derivatives with partial structure of β1-3/1-6 glucan, using monomeric units for the formation of β1-3 and β1-6 glucosidic linkages.
Tomoya Ota; Wataru Saburi; Shiro Komba; Haruhide Mori
Bioscience, biotechnology, and biochemistry, 05 Jul. 2023, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, β1-3/1-6 Glucans, known for their diverse structures, comprise a β1-3-linked main chain and β1-6-linked short branches. Laminarin, a β1-3/1-6 glucan extracted from brown seaweed, for instance, includes β1-6 linkages even in the main chain. The diverse structures provide various beneficial functions of the glucan. To investigate the relationship between structure and functionality, and to enable the characterization of β1-3/1-6 glucan-metabolizing enzymes, oligosaccharides containing exact structures of β1-3/1-6 glucans are required. We synthesized the monomeric units for the synthesis of β1-3/1-6 mixed-linked glucooligosaccharides. 2-(Trimethylsilyl)ethyl 2-O-benzoyl-4,6-O-benzylidene-β-d-glucopyranoside served as an acceptor in the formation of β1-3 linkages. Phenyl 2-O-benzoyl-4,6-O-benzylidene-3-O-(tert-butyldiphenylsilyl)-1-thio-β-d-glucopyranoside and phenyl 2,3-di-O-benzoyl-4,6-di-O-levulinyl-1-thio-β-d-glucopyranoside acted as donors, synthesizing acceptors suitable for the formation of β1-3- and β1-6-linkages, respectively. These were used to synthesize a derivative of Glcβ1-6Glcβ1-3Glcβ1-3Glc, demonstrating that the proposed route can be applied to synthesize the main chain of β-glucan, with the inclusion of both β1-3 and β1-6 linkages.

Structural insights into the substrate specificity and activity of a novel mannose 2-epimerase from Runella slithyformis.
Hang Wang; Xiaomei Sun; Wataru Saburi; Saki Hashiguchi; Jian Yu; Toyoyuki Ose; Haruhide Mori; Min Yao
Acta crystallographica. Section D, Structural biology, 79, Pt 7, 585, 595, 01 Jul. 2023, [Peer-reviewed], [International Magazine]
English, Scientific journal, Mannose 2-epimerase (ME), a member of the acylglucosamine 2-epimerase (AGE) superfamily that catalyzes epimerization of D-mannose and D-glucose, has recently been characterized to have potential for D-mannose production. However, the substrate-recognition and catalytic mechanism of ME remains unknown. In this study, structures of Runella slithyformis ME (RsME) and its D254A mutant [RsME(D254A)] were determined in their apo forms and as intermediate-analog complexes [RsME-D-glucitol and RsME(D254A)-D-glucitol]. RsME possesses the (α/α)6-barrel of the AGE superfamily members but has a unique pocket-covering long loop (loopα7-α8). The RsME-D-glucitol structure showed that loopα7-α8 moves towards D-glucitol and closes the active pocket. Trp251 and Asp254 in loopα7-α8 are only conserved in MEs and interact with D-glucitol. Kinetic analyses of the mutants confirmed the importance of these residues for RsME activity. Moreover, the structures of RsME(D254A) and RsME(D254A)-D-glucitol revealed that Asp254 is vital for binding the ligand in a correct conformation and for active-pocket closure. Docking calculations and structural comparison with other 2-epimerases show that the longer loopα7-α8 in RsME causes steric hindrance upon binding to disaccharides. A detailed substrate-recognition and catalytic mechanism for monosaccharide-specific epimerization in RsME has been proposed.

Identification and characterization of extracellular GH3 β-glucosidase from the pink snow mold fungus, Microdochium nivale.
Tomoya Ota; Wataru Saburi; Linda Elizabeth Jewell; Tom Hsiang; Ryozo Imai; Haruhide Mori
Bioscience, biotechnology, and biochemistry, 87, 7, 707, 716, 23 Jun. 2023, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Glycoside hydrolase family 3 (GH3) β-glucosidase exists in many filamentous fungi. In phytopathogenic fungi, it is involved in fungal growth and pathogenicity. Microdochium nivale is a severe phytopathogenic fungus of grasses and cereals and is the causal agent of pink snow mold, but its β-glucosidase has not been identified. In this study, a GH3 β-glucosidase of M. nivale (MnBG3A) was identified and characterized. Among various p-nitrophenyl β-glycosides, MnBG3A showed activity on d-glucoside (pNP-Glc) and slight activity on d-xyloside. In the pNP-Glc hydrolysis, substrate inhibition occurred (Kis = 1.6 m m), and d-glucose caused competitive inhibition (Ki = 0.5 m m). MnBG3A acted on β-glucobioses with β1-3, -6, -4, and -2 linkages, in descending order of kcat/Km. In contrast, the regioselectivity for newly formed products was limited to β1-6 linkage. MnBG3A has similar features to those of β-glucosidases from Aspergillus spp., but higher sensitivity to inhibitory effects.

Alteration of Substrate Specificity and Transglucosylation Activity of GH13_31 α-Glucosidase from Bacillus sp. AHU2216 through Site-Directed Mutagenesis of Asn258 on β→α Loop 5
Waraporn Auiewiriyanukul; Wataru Saburi; Tomoya Ota; Jian Yu; Koji Kato; Min Yao; Haruhide Mori
Molecules, 28, 7, 3109, 3109, MDPI AG, 30 Mar. 2023, [Peer-reviewed], [Invited], [Last author, Corresponding author]
Scientific journal, α-Glucosidase catalyzes the hydrolysis of α-d-glucosides and transglucosylation. Bacillus sp. AHU2216 α-glucosidase (BspAG13_31A), belonging to the glycoside hydrolase family 13 subfamily 31, specifically cleaves α-(1→4)-glucosidic linkages and shows high disaccharide specificity. We showed previously that the maltose moiety of maltotriose (G3) and maltotetraose (G4), covering subsites +1 and +2 of BspAG13_31A, adopts a less stable conformation than the global minimum energy conformation. This unstable d-glucosyl conformation likely arises from steric hindrance by Asn258 on β→α loop 5 of the catalytic (β/α)8-barrel. In this study, Asn258 mutants of BspAG13_31A were enzymatically and structurally analyzed. N258G/P mutations significantly enhanced trisaccharide specificity. The N258P mutation also enhanced the activity toward sucrose and produced erlose from sucrose through transglucosylation. N258G showed a higher specificity to transglucosylation with p-nitrophenyl α-d-glucopyranoside and maltose than the wild type. E256Q/N258G and E258Q/N258P structures in complex with G3 revealed that the maltose moiety of G3 bound at subsites +1 and +2 adopted a relaxed conformation, whereas a less stable conformation was taken in E256Q. This structural difference suggests that stabilizing the G3 conformation enhances trisaccharide specificity. The E256Q/N258G-G3 complex formed an additional hydrogen bond between Met229 and the d-glucose residue of G3 in subsite +2, and this interaction may enhance transglucosylation.

Function and structure of Lacticaseibacillus casei GH35 β-galactosidase LBCZ_0230 with high hydrolytic activity to lacto-N-biose I and galacto-N-biose
Saburi Wataru; Tomoya Ota; Koji Kato; Takayoshi Tagami; Keitaro Yamashita; Min Yao; Haruhide Mori
Journal of Applied Glycoscience, 70, 2, 43, 52, The Japanese Society of Applied Glycoscience, 11 Mar. 2023, [Peer-reviewed], [Last author]
Scientific journal

Substrate specificity of glycoside hydrolase family 1 β-glucosidase AtBGlu42 from Arabidopsis thaliana and its molecular mechanism.
Shu Horikoshi; Wataru Saburi; Jian Yu; Hideyuki Matsuura; James R Ketudat Cairns; Min Yao; Haruhide Mori
Bioscience, biotechnology, and biochemistry, 86, 2, 231, 245, 24 Jan. 2022, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Plants possess many glycoside hydrolase family 1 (GH1) β-glucosidases, which physiologically function in cell wall metabolism and activation of bioactive substances, but most remain uncharacterized. One GH1 isoenzyme AtBGlu42 in Arabidopsis thaliana has been identified to hydrolyze scopolin using the gene deficient plants, but no enzymatic properties were obtained. Its sequence similarity to another functionally characterized enzyme Os1BGlu4 in rice suggests that AtBGlu42 also acts on oligosaccharides. Here, we show that the recombinant AtBGlu42 possesses high kcat/Km not only on scopolin, but also on various β-glucosides, cellooligosaccharides, and laminarioligosaccharides. Of the cellooligosaccharides, cellotriose was the most preferred. The crystal structure, determined at 1.7 Å resolution, suggests that Arg342 gives unfavorable binding to cellooligosaccharides at subsite +3. The mutants R342Y and R342A showed the highest preference on cellotetraose or cellopentaose with increased affinities at subsite +3, indicating that the residues at this position have an important role for chain length specificity.

A practical approach to producing isomaltomegalosaccharide using dextran dextrinase from Gluconobacter oxydans ATCC 11894.
Weeranuch Lang; Yuya Kumagai; Juri Sadahiro; Wataru Saburi; Rakrudee Sarnthima; Takayoshi Tagami; Masayuki Okuyama; Haruhide Mori; Nobuo Sakairi; Doman Kim; Atsuo Kimura
Applied microbiology and biotechnology, 106, 2, 689, 698, Jan. 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Dextran dextrinase (DDase) catalyzes formation of the polysaccharide dextran from maltodextrin. During the synthesis of dextran, DDase also generates the beneficial material isomaltomegalosaccharide (IMS). The term megalosaccharide is used for a saccharide having DP = 10-100 or 10-200 (DP, degree of polymerization). IMS is a chimeric glucosaccharide comprising α-(1 → 6)- and α-(1 → 4)-linked portions at the nonreducing and reducing ends, respectively, in which the α-(1 → 4)-glucosyl portion originates from maltodextrin of the substrate. In this study, IMS was produced by a practical approach using extracellular DDase (DDext) or cell surface DDase (DDsur) of Gluconobacter oxydans ATCC 11894. DDsur was the original form, so we prepared DDext via secretion from intact cells by incubating with 0.5% G6/G7 (maltohexaose/maltoheptaose); this was followed by generation of IMS from various concentrations of G6/G7 substrate at different temperatures for 96 h. However, IMS synthesis by DDext was limited by insufficient formation of α-(1 → 6)-glucosidic linkages, suggesting that DDase also catalyzes elongation of α-(1 → 4)-glucosyl chain. For production of IMS using DDsur, intact cells bearing DDsur were directly incubated with 20% G6/G7 at 45 °C by optimizing conditions such as cell concentration and agitation efficiency, which resulted in generation of IMS (average DP = 14.7) with 61% α-(1 → 6)-glucosyl content in 51% yield. Increases in substrate concentration and agitation efficiency were found to decrease dextran formation and increase IMS production, which improved the reaction conditions for DDext. Under modified conditions (20% G6/G7, agitation speed of 100 rpm at 45 °C), DDext produced IMS (average DP = 14.5) with 65% α-(1 → 6)-glucosyl content in a good yield of 87%. KEY POINTS: • Beneficial IMS was produced using thermostabilized DDase. • Optimum conditions for reduced dextran formation were successfully determined. • A practical approach was established to provide IMS with a great yield of 87%.

Discovery of solabiose phosphorylase and its application for enzymatic synthesis of solabiose from sucrose and lactose
Wataru Saburi; Takanori Nihira; Hiroyuki Nakai; Motomitsu Kitaoka; Haruhide Mori
Scientific Reports, 12, 1, Springer Science and Business Media LLC, Jan. 2022, [Peer-reviewed], [Last author]
Scientific journal, Glycoside phosphorylases (GPs), which catalyze the reversible phosphorolysis of glycosides, are promising enzymes for the efficient production of glycosides. Various GPs with new catalytic activities are discovered from uncharacterized proteins phylogenetically distant from known enzymes in the past decade. In this study, we characterized Paenibacillus borealis PBOR_28850 protein, belonging to glycoside hydrolase family 94. Screening of acceptor substrates for reverse phosphorolysis, in which α-d-glucose 1-phosphate was used as the donor substrate, revealed that the recombinant PBOR_28850 produced in Escherichia coli specifically utilized d-galactose as an acceptor and produced solabiose (β-d-Glcp-(1 → 3)-d-Gal). This indicates that PBOR_28850 is a new GP, solabiose phosphorylase. PBOR_28850 catalyzed the phosphorolysis and synthesis of solabiose through a sequential bi-bi mechanism involving the formation of a ternary complex. The production of solabiose from lactose and sucrose has been established. Lactose was hydrolyzed to d-galactose and d-glucose by β-galactosidase. Phosphorolysis of sucrose and synthesis of solabiose were then coupled by adding sucrose, sucrose phosphorylase, and PBOR_28850 to the reaction mixture. Using 210 mmol lactose and 280 mmol sucrose, 207 mmol of solabiose was produced. Yeast treatment degraded the remaining monosaccharides and sucrose without reducing solabiose. Solabiose with a purity of 93.7% was obtained without any chromatographic procedures.

A Ubiquitously Expressed UDP-Glucosyltransferase, UGT74J1, Controls Basal Salicylic Acid Levels in Rice
Daisuke Tezuka; Hideyuki Matsuura; Wataru Saburi; Haruhide Mori; Ryozo Imai
Plants, 10, 9, 1875, 1875, MDPI AG, 10 Sep. 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, Salicylic acid (SA) is a phytohormone that regulates a variety of physiological and developmental processes, including disease resistance. SA is a key signaling component in the immune response of many plant species. However, the mechanism underlying SA-mediated immunity is obscure in rice (Oryza sativa). Prior analysis revealed a correlation between basal SA level and blast resistance in a range of rice varieties. This suggested that resistance might be improved by increasing basal SA level. Here, we identified a novel UDP-glucosyltransferase gene, UGT74J1, which is expressed ubiquitously throughout plant development. Mutants of UGT74J1 generated by genome editing accumulated high levels of SA under non-stressed conditions, indicating that UGT74J1 is a key enzyme for SA homeostasis in rice. Microarray analysis revealed that the ugt74j1 mutants constitutively overexpressed a set of pathogenesis-related (PR) genes. An inoculation assay demonstrated that these mutants had increased resistance against rice blast, but they also exhibited stunted growth phenotypes. To our knowledge, this is the first report of a rice mutant displaying SA overaccumulation.

Preliminary evaluation of colorimetric and HPLC-based methods for quantifying β-(1→4)-mannobiose in a crude material
Kensuke Fukui; Wataru Saburi; Masahisa Ibuki; Kazunobu Tsumura; Haruhide Mori
Food Science and Technology Research, 27, 2, 249, 257, Japanese Society for Food Science and Technology, 22 May 2021, [Peer-reviewed]
Scientific journal

[Review] Functions of Hydrolases, Phosphorylases, and Isomerases Acting on Carbohydrates, and their Application
Haruhide Mori
Bulletin of Applied Glycoscience, 10, 3, 165, 174, The Japanese Society of Applied Glycoscience, 20 Aug. 2020, [Peer-reviewed], [Invited], [Lead author, Last author, Corresponding author]
Japanese, Scientific journal

Efficient one-pot enzymatic synthesis of trehalose 6-phosphate using GH65 α-glucoside phosphorylases.
Yodai Taguchi; Wataru Saburi; Ryozo Imai; Haruhide Mori
Carbohydrate research, 488, 107902, 107902, Feb. 2020, [Peer-reviewed], [Last author], [International Magazine]
English, Trehalose 6-phosphate (Tre6P) is an important intermediate for trehalose biosynthesis. Recent researches have revealed that Tre6P is an endogenous signaling molecule that regulates plant development and stress responses. The necessity of Tre6P in physiological studies is expected to be increasing. To achieve the cost-effective production of Tre6P, a novel approach is required. In this study, we utilized trehalose 6-phosphate phosphorylase (TrePP) from Lactococcus lactis to produce Tre6P. In the reverse phosphorolysis by the TrePP, 91.9 mM Tre6P was produced from 100 mM β-glucose 1-phosphate (β-Glc1P) and 100 mM glucose 6-phosphate (Glc6P). The one-pot reaction of TrePP and maltose phosphorylase (MP) enabled production of 65 mM Tre6P from 100 mM maltose, 100 mM Glc6P, and 20 mM inorganic phosphate. Addition of β-phosphoglucomutase to this reaction produced Glc6P from β-Glc1P and thus reduced requirement of Glc6P as a starting material. Within the range of 20-469 mM inorganic phosphate tested, the 54 mM concentration yielded the highest amount of Tre6P (33 mM). Addition of yeast increased the yield because of its glucose consumption. Finally, from 100 mmol maltose and 60 mmol inorganic phosphate, we successfully achieved production of 37.5 mmol Tre6P in a one-pot reaction (100 mL), and 9.4 g Tre6P dipotassium salt was obtained.

Biochemical characteristics of maltose phosphorylase MalE from Bacillus sp. AHU2001 and chemoenzymatic synthesis of oligosaccharides by the enzyme.
Gao Y; Saburi W; Taguchi Y; Mori H
Bioscience, biotechnology, and biochemistry, 1, 13, Jul. 2019, [Peer-reviewed]

Enzymatic characteristics of D-mannose 2-epimerase, a new member of the acylglucosamine 2-epimerase superfamily.
Saburi W; Sato S; Hashiguchi S; Muto H; Iizuka T; Mori H
Applied microbiology and biotechnology, Jun. 2019, [Peer-reviewed]

The rice ethylene response factor OsERF83 positively regulates disease resistance to Magnaporthe oryzae.
Tezuka D; Kawamata A; Kato H; Saburi W; Mori H; Imai R
Plant Physiol Biochem, 135, 263, 271, Elsevier BV, Feb. 2019, [Peer-reviewed]
English, Scientific journal

A Transposon Mutagenesis System for Bifidobacterium longum subsp. longum Based on an IS3 Family Insertion Sequence, ISBlo11.
Mikiyasu Sakanaka; Shingo Nakakawaji; Shin Nakajima; Satoru Fukiya; Arisa Abe; Wataru Saburi; Haruhide Mori; Atsushi Yokota
Applied and environmental microbiology, 84, 17, e00824-18, 01 Sep. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Bifidobacteria are a major component of the intestinal microbiota in humans, particularly breast-fed infants. Therefore, elucidation of the mechanisms by which these bacteria colonize the intestine is desired. One approach is transposon mutagenesis, a technique currently attracting much attention because, in combination with next-generation sequencing, it enables exhaustive identification of genes that contribute to microbial fitness. We now describe a transposon mutagenesis system for Bifidobacterium longum subsp. longum 105-A (JCM 31944) based on ISBlo11, a native IS3 family insertion sequence. To build this system, xylose-inducible or constitutive bifidobacterial promoters were tested to drive the expression of full-length or a truncated form at the N terminus of the ISBlo11 transposase. An artificial transposon plasmid, pBFS12, in which ISBlo11 terminal inverted repeats are separated by a 3-bp spacer, was also constructed to mimic the transposition intermediate of IS3 elements. The introduction of this plasmid into a strain expressing transposase resulted in the insertion of the plasmid with an efficiency of >103 CFU/μg DNA. The plasmid targets random 3- to 4-bp sequences, but with a preference for noncoding regions. This mutagenesis system also worked at least in B. longum NCC2705. Characterization of a transposon insertion mutant revealed that a putative α-glucosidase mediates palatinose and trehalose assimilation, demonstrating the suitability of transposon mutagenesis for loss-of-function analysis. We anticipate that this approach will accelerate functional genomic studies of B. longum subsp. longumIMPORTANCE Several hundred species of bacteria colonize the mammalian intestine. However, the genes that enable such bacteria to colonize and thrive in the intestine remain largely unexplored. Transposon mutagenesis, combined with next-generation sequencing, is a promising tool to comprehensively identify these genes but has so far been applied only to a small number of intestinal bacterial species. In this study, a transposon mutagenesis system was established for Bifidobacterium longum subsp. longum, a representative health-promoting Bifidobacterium species. The system enables the identification of genes that promote colonization and survival in the intestine and should help illuminate the physiology of this species.

Function and structure of GH13_31 α-glucosidase with high α-(1→4)-glucosidic linkage specificity and transglucosylation activity.
Auiewiriyanukul W; Saburi W; Kato K; Yao M; Mori H
FEBS Lett, 592, 13, 2268, 2281, Jul. 2018, [Peer-reviewed]

Biochemical and structural characterization of Marinomonas mediterranea D-mannose isomerase Marme_2490 phylogenetically distant from known enzymes
Wataru Saburi; Nongluck Jaito; Koji Kato; Yuka Tanaka; Min Yao; Haruhide Mori
Biochimie, 144, 63, 73, Elsevier B.V., 01 Jan. 2018, [Peer-reviewed]
English, Scientific journal

Effects of mutation of Asn694 in Aspergillus niger alpha-glucosidase on hydrolysis and transglucosylation
Min Ma; Masayuki Okuyama; Megumi Sato; Takayoshi Tagami; Patcharapa Klahan; Yuya Kumagai; Haruhide Mori; Atsuo Kimura
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 101, 16, 6399, 6408, Aug. 2017, [Peer-reviewed]
English, Scientific journal

Elucidation of the biosynthetic pathway of cis-jasmone in Lasiodiplodia theobromae
Ryo Matsui; Naruki Amano; Kosaku Takahashi; Yodai Taguchi; Wataru Saburi; Hideharu Mori; Norio Kondo; Kazuhiko Matsuda; Hideyuki Matsuura
SCIENTIFIC REPORTS, 7, 1, 6688, Jul. 2017, [Peer-reviewed]
English, Scientific journal

β-マンナン分解に寄与するセロビオース２-エピメラーゼとβ-マンノシドホスホリラーゼの構造と機能
佐分利 亘; 加藤 公児; 姚 閔; 松井 博和; 森 春英
応用糖質科学, 7, 2, 69, 75, May 2017, [Peer-reviewed], [Invited]
Japanese, Scientific journal

Efficient synthesis of alpha-galactosyl oligosaccharides using a mutant Bacteroides thetaiotaomicron retaining alpha-galactosidase (BtGH97b)
Masayuki Okuyama; Kana Matsunaga; Ken-ichi Watanabe; Keitaro Yamashita; Takayoshi Tagami; Asako Kikuchi; Min Ma; Patcharapa Klahan; Haruhide Mori; Min Yao; Atsuo Kimura
FEBS JOURNAL, 284, 5, 766, 783, Mar. 2017, [Peer-reviewed]
English, Scientific journal

Evaluation of acceptor selectivity of Lactococcus lactis ssp lactis trehalose 6-phosphate phosphorylase in the reverse phosphorolysis and synthesis of a new sugar phosphate
Yodai Taguchi; Wataru Saburi; Ryozo Imai; Haruhide Mori
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 81, 8, 1512, 1519, 2017, [Peer-reviewed]
English, Scientific journal

Kinetic properties and substrate inhibition of -galactosidase from Aspergillus niger
Julan Liao; Masayuki Okuyama; Keigo Ishihara; Yoshinori Yamori; Shigeo Iki; Takayoshi Tagami; Haruhide Mori; Seiya Chiba; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 80, 9, 1747, 1752, Sep. 2016, [Peer-reviewed]
English, Scientific journal

alpha-Glucosidases and alpha-1,4-glucan lyases: structures, functions, and physiological actions
Masayuki Okuyama; Wataru Saburi; Haruhide Mori; Atsuo Kimura
CELLULAR AND MOLECULAR LIFE SCIENCES, 73, 14, 2727, 2751, Jul. 2016, [Peer-reviewed]
English, Scientific journal

Structural insights into the difference in substrate recognition of two mannoside phosphorylases from two GH130 subfamilies
Yuxin Ye; Wataru Saburi; Rei Odaka; Koji Kato; Naofumi Sakurai; Keisuke Komoda; Mamoru Nishimoto; Motomitsu Kitaoka; Haruhide Mori; Min Yao
FEBS LETTERS, 590, 6, 828, 837, Mar. 2016, [Peer-reviewed]
English, Scientific journal

Purification and characterization of a chloride ion-dependent alpha-glucosidase from the midgut gland of Japanese scallop (Patinopecten yessoensis)
Yasushi Masuda; Masayuki Okuyama; Takahisa Iizuka; Hiroyuki Nakai; Wataru Saburi; Taro Fukukawa; Janjira Maneesan; Takayoshi Tagami; Tetsushi Naraoka; Haruhide Mori; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 80, 3, 479, 485, Mar. 2016, [Peer-reviewed]
English, Scientific journal

Structural and biochemical studies of plant α-glucosidases with a series of long-chain inhibitors.
Tagami T; Yamashita K; Okuyama M; Mori H; Yao M; Kimura A
Bull Appl Glycosci, 6, 2, 103, 108, 2016, [Peer-reviewed]
Japanese, Scientific journal

Supplemental epilactose prevents metabolic disorders through uncoupling protein-1 induction in the skeletal muscle of mice fed high-fat diets
Yuki Murakami; Teruyo Ojima-Kato; Wataru Saburi; Haruhide Mori; Hirokazu Matsui; Soichi Tanabe; Takuya Suzuki
BRITISH JOURNAL OF NUTRITION, 114, 11, 1774, 1783, Dec. 2015, [Peer-reviewed]
English, Scientific journal

Identification of rice Os4BGlu13 as a beta-glucosidase which hydrolyzes gibberellin A4 1-O-beta-D-glucosyl ester, in addition to tuberonic acid glucoside and salicylic acid derivative glucosides
Yanling Hua; Watsamon Ekkhara; Sompong Sansenya; Chantragan Srisomsap; Sittiruk Roytrakul; Wataru Saburi; Ryosuke Takeda; Hideyuki Matsuura; Haruhide Mori; James R. Ketudat Cairns
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 583, 36, 46, Oct. 2015, [Peer-reviewed]
English, Scientific journal

A Single-Nucleotide Polymorphism in an Endo-1,4-beta-Glucanase Gene Controls Seed Coat Permeability in Soybean
Seong-Jin Jang; Masako Sato; Kei Sato; Yutaka Jitsuyama; Kaien Fujino; Haruhide Mori; Ryoji Takahashi; Eduardo R. Benitez; Baohui Liu; Tetsuya Yamada; Jun Abe
PLOS ONE, 10, 6, e0128527, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Functional reassignment of Cellvibrio vulgaris EpiA to cellobiose 2-epimerase and an evaluation of the biochemical functions of the 4-O-beta-d-mannosyl-d-glucose phosphorylase-like protein, UnkA
Wataru Saburi; Yuka Tanaka; Hirohiko Muto; Sota Inoue; Rei Odaka; Mamoru Nishimoto; Motomitsu Kitaoka; Haruhide Mori
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 79, 6, 969, 977, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Structural analysis of the alpha-glucosidase HaG provides new insights into substrate specificity and catalytic mechanism
Xing Shen; Wataru Saburi; Zuoqi Gai; Koji Kato; Teruyo Ojima-Kato; Jian Yu; Keisuke Komoda; Yusuke Kido; Hirokazu Matsui; Haruhide Mori; Min Yao
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 71, Pt 6, 1382, 1391, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Structural elements responsible for the glucosidic linkage-selectivity of a glycoside hydrolase family 13 exo-glucosidase
Wataru Saburi; Hiroaki Rachi-Otsuka; Hironori Hondoh; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
FEBS LETTERS, 589, 7, 865, 869, Mar. 2015, [Peer-reviewed]
English, Scientific journal

Structural insights into the catalytic reaction that is involved in the reorientation of Trp238 at the substrate-binding site in GH13 dextran glucosidase
Momoko Kobayashi; Wataru Saburi; Daichi Nakatsuka; Hironori Hondoh; Koji Kato; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura; Min Yao
FEBS LETTERS, 589, 4, 484, 489, Feb. 2015, [Peer-reviewed]
English, Scientific journal

UDP-Glc Independent Glucosyltransferase toward Jasmonic acid Derivatives and its Biological Role.
Takematsu Tomonori; Seto Yoshiya; Miyazawa Yoshiro; Wakuta Shinji; Ogihara Tsuyoshi; Saburi Wataru; Mori Haruhide; Takahashi Kosaku; Matsuura Hideyuki
Symposium on the Chemistry of Natural Products, symposium papers, 57, Oral14, Symposium on the Chemistry of Natural Products Steering Committee, 2015
Japanese,

Plants are sessile organisms and are unable to avoid environmental stresses by changing their habitats. Therefore, plants have developed unique and sophisticated responding systems. It has been generally accepted that plant use plant hormones to give actions toward (against) environmental changes and stress. Among of the hormones, jasmonic acid(s) has pivotal roles to perform the responses. In recent years, not only the activation of JA pathway but also the deactivations of active form JA are being paid a lot of attentions such as oxidations and glucosylations.

In previous our paper [1], we reported that Os SGT, putative salicylic acid glucosyltransferase, transferred glucosyl moiety toward 12-OHJA to afford 12-OGlcJA, and its mRNA was induced by wounding stress and JA and SA treatments. In the course of that study, we also found UDP-Glc independent glucosyl transferase activity to give preferably 12-OGlcJA in the crude extract of rice cell culture using octyl glucoside as donor molecule for supplying glucosyl moiety. There are few reports of the finding on UDP-Glc independent glucosyltransferase protein, and to our best knowledge, a report has been published by Matsuba et al. [2]. In this presentation we discuss elucidation of UDP-Glc independent glucosyl transferase toward 12-OHJA and 12-OHJA-Ile

[1] Seto Y. et al., Phytochemistry, 70, 370-379 (2009).

[2] Matsuba Y. et al.,Plant Cell,22, 3374-3389 (2010).

Identity of the two dextran dextrinases produced by Gluconobacter oxydans ATCC 11894 and its localization change depending on the cell growth.
Sadahiro J; Mori H; Saburi W; Okuyama M; Kimura A
Biochem Biophys Res Commun, 456, 1, 500, 505, 2015, [Peer-reviewed]
English, Scientific journal

Biochemical properties and substrate recognition mechanism of GH31 alpha-glucosidase from Bacillus sp AHU 2001 with broad substrate specificity
Wataru Saburi; Masayuki Okuyama; Yuya Kumagai; Atsuo Kimura; Haruhide Mori
BIOCHIMIE, 108, 140, 148, Jan. 2015, [Peer-reviewed]
English, Scientific journal

Structural Advantage of Sugar Beet alpha-Glucosidase to Stabilize the Michaelis Complex with Long-chain Substrate
Takayoshi Tagami; Keitaro Yamashita; Masayuki Okuyama; Haruhide Mori; Min Yao; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 290, 3, 1796, 1803, Jan. 2015, [Peer-reviewed]
English, Scientific journal

Different molecular complexity of linear-isomaltomegalosaccharides and beta-cyclodextrin on enhancing solubility of azo dye ethyl red: Towards dye biodegradation
Weeranuch Lang; Yuya Kumagai; Juri Sadahiro; Janjira Maneesan; Masayuki Okuyama; Haruhide Mori; Nobuo Sakairi; Atsuo Kimura
BIORESOURCE TECHNOLOGY, 169, 518, 524, Oct. 2014, [Peer-reviewed]
English, Scientific journal

Catalytic role of the calcium ion in GH97 inverting glycoside hydrolase
Masayuki Okuyama; Takuya Yoshida; Hironori Hondoh; Haruhide Mori; Min Yao; Atsuo Kimura
FEBS LETTERS, 588, 17, 3213, 3217, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Enhancement of hydrolytic activity of thermophilic alkalophilic alpha-amylase from Bacillus sp AAH-31 through optimization of amino acid residues surrounding the substrate binding site
Naoya Tamamura; Wataru Saburi; Atsushi Mukai; Naoki Morimoto; Toshihiko Takehana; Seiji Koike; Hirokazu Matsui; Haruhide Mori
BIOCHEMICAL ENGINEERING JOURNAL, 86, 8, 15, May 2014, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary X-ray crystallographic analysis of α-glucosidase HaG from Halomonas sp. strain H11.
Shen X; Saburi W; Gai ZQ; Komoda K; Yu J; Ojima-Kato T; Kido Y; Matsui H; Mori H; Yao M
Acta crystallographica. Section F, Structural biology communications, 70, Pt 4, 464, 466, Apr. 2014, [Peer-reviewed], [International Magazine]
English, Scientific journal, The α-glucosidase HaG from the halophilic bacterium Halomonas sp. strain H11 catalyzes the hydrolysis of the glucosidic linkage at the nonreducing end of α-glucosides, such as maltose and sucrose, to release α-glucose. Based on its amino-acid sequence, this enzyme is classified as a member of glycoside hydrolase family 13. HaG has three unique characteristics: (i) a very narrow substrate specificity, almost exclusively hydrolyzing disaccharides; (ii) activation by monovalent cations, such as K(+), Rb(+), Cs(+) and NH4(+); and (iii) high transfer activity of the glucose moiety to the OH group of low-molecular-weight compounds, including glycerol and 6-gingerol. Crystallographic studies have been performed in order to understand these special features. An expression vector was constructed and recombinant HaG protein was overexpressed, purified and crystallized. A data set to 2.15 Å resolution was collected and processed. The crystal belonged to space group P212121, with unit-cell parameters a = 60.2, b = 119.2, c = 177.2 Å. The structure has been determined by molecular replacement using the isomaltulose synthase PalI as the search model (PDB entry 1m53).

Structural insights into the epimerization of β-1,4-linked oligosaccharides catalyzed by cellobiose 2-epimerase, the sole enzyme epimerizing non-anomeric hydroxyl groups of unmodified sugars.
Fujiwara T; Saburi W; Matsui H; Mori H; Yao M
The Journal of biological chemistry, 289, 6, 3405, 3415, American Society for Biochemistry and Molecular Biology (ASBMB), Feb. 2014, [Peer-reviewed]
English, Cellobiose 2-epimerase (CE) reversibly converts d-glucose residues into d-mannose residues at the reducing end of unmodified β1,4-linked oligosaccharides, including β-1,4-mannobiose, cellobiose, and lactose. CE is responsible for conversion of β1,4-mannobiose to 4-O-β-d-mannosyl-d-glucose in mannan metabolism. However, the detailed catalytic mechanism of CE is unclear due to the lack of structural data in complex with ligands. We determined the crystal structures of halothermophile Rhodothermus marinus CE (RmCE) in complex with substrates/products or intermediate analogs, and its apo form. The structures in complex with the substrates/products indicated that the residues in the β5-β6 loop as well as those in the inner six helices form the catalytic site. Trp-322 and Trp-385 interact with reducing and non-reducing end parts of these ligands, respectively, by stacking interactions. The architecture of the catalytic site also provided insights into the mechanism of reversible epimerization. His-259 abstracts the H2 proton of the d-mannose residue at the reducing end, and consistently forms the cis-enediol intermediate by facilitated depolarization of the 2-OH group mediated by hydrogen bonding interaction with His-200. His-390 subsequently donates the proton to the C2 atom of the intermediate to form a d-glucose residue. The reverse reaction is mediated by these three histidines with the inverse roles of acid/base catalysts. The conformation of cellobiitol demonstrated that the deprotonation/reprotonation step is coupled with rotation of the C2-C3 bond of the open form of the ligand. Moreover, it is postulated that His-390 is closely related to ring opening/closure by transferring a proton between the O5 and O1 atoms of the ligand.

Production of 1,5-anhydro-D-fructose by an α-glucosidase belonging to glycoside hydrolase family 31.
Maneesan J; Matsuura H; Tagami T; Mori H; Kimura A
Biosci Biotechnol Biochem, 78, 12, 2064, 2068, 2014, [Peer-reviewed]
English, Scientific journal

Characterization of a thermophilic 4-O-β-D-mannosyl-D-glucose phosphorylase from Rhodothermus marinus
Nongluck Jaito; Wataru Saburi; Rei Odaka; Yusuke Kido; Ken Hamura; Mamoru Nishimoto; Motomitsu Kitaoka; Hirokazu Matsui; Haruhide Mori
Bioscience, Biotechnology and Biochemistry, 78, 2, 263, 270, Taylor & Francis, 01 Jan. 2014
English

Biodecolorization of a food azo dye by the deep sea Dermacoccus abyssi MT1.1(T) strain from the Mariana Trench
Weeranuch Lang; Sarote Sirisansaneeyakul; Ligia O. Martins; Lukana Ngiwsara; Nobuo Sakairi; Wasu Pathom-aree; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
JOURNAL OF ENVIRONMENTAL MANAGEMENT, 132, 155, 164, Jan. 2014, [Peer-reviewed]
English, Scientific journal

Replacement of the Catalytic Nucleophile Aspartyl Residue of Dextran Glucosidase by Cysteine Sulfinate Enhances Transglycosylation Activity
Wataru Saburi; Momoko Kobayashi; Haruhide Mori; Masayuki Okuyama; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 288, 44, 31670, 31677, Nov. 2013, [Peer-reviewed]
English, Scientific journal

Characterization of Ruminococcus albus cellodextrin phosphorylase and identification of a key phenylalanine residue for acceptor specificity and affinity to the phosphate group.
Sawano T; Saburi W; Hamura K; Matsui H; Mori H
The FEBS journal, 280, 18, 4463, 4473, Wiley-Blackwell, Sep. 2013, [Peer-reviewed]
English, Ruminococcus albus has the ability to intracellularly degrade cello-oligosaccharides primarily via phosphorolysis. In this study, the enzymatic characteristics of R. albus cellodextrin phosphorylase (RaCDP), which is a member of glycoside hydrolase family 94, was investigated. RaCDP catalyzes the phosphorolysis of cellotriose through an ordered 'bi bi' mechanism in which cellotriose binds to RaCDP before inorganic phosphate, and then cellobiose and glucose 1-phosphate (Glc1P) are released in that order. Among the cello-oligosaccharides tested, RaCDP had the highest phosphorolytic and synthetic activities towards cellohexaose and cellopentaose, respectively. RaCDP successively transferred glucosyl residues from Glc1P to the growing cello-oligosaccharide chain, and insoluble cello-oligosaccharides comprising a mean of eight residues were produced. Sophorose, laminaribiose, β-1,4-xylobiose, β-1,4-mannobiose and cellobiitol served as acceptors for RaCDP. RaCDP had very low affinity for phosphate groups in both the phosphorolysis and synthesis directions. A sequence comparison revealed that RaCDP has Gln at position 646 where His is normally conserved in the phosphate binding sites of related enzymes. A Q646H mutant showed approximately twofold lower apparent Km values for inorganic phosphate and Glc1P than the wild-type. RaCDP has Phe at position 633 corresponding to Tyr and Val in the +1 subsites of cellobiose phosphorylase and N,N′-diacetylchitobiose phosphorylase, respectively. A F633Y mutant showed higher preference for cellobiose over β-1,4-mannobiose as an acceptor substrate in the synthetic reaction than the wild-type. Furthermore, the F633Y mutant showed 75- and 1100-fold lower apparent Km values for inorganic phosphate and Glc1P, respectively, in phosphorolysis and synthesis of cellotriose.

Modulation of acceptor specificity of Ruminococcus albus cellobiose phosphorylase through site-directed mutagenesis.
Hamura K; Saburi W; Matsui H; Mori H
Carbohydrate research, 379, 21, 25, Elsevier sci ltd, Sep. 2013, [Peer-reviewed]
English, Cellobiose phosphorylase (EC 2.4.1.20, CBP) catalyzes the reversible phosphorolysis of cellobiose to alpha-D-glucose 1-phosphate (Glc1P) and D-glucose. Cys485, Tyr648, and Glu653 of CBP from Ruminococcus albus, situated at the +1 subsite, were mutated to modulate acceptor specificity. C485A, Y648F, and Y648V were active enough for analysis. Their acceptor specificities were compared with the wild type based on the apparent kinetic parameters determined in the presence of 10 mM Glc1P. C485A showed higher preference for D-glucosamine than the wild type. Apparent k(cat)/K-m values of Y648F for D-mannose and 2-deoxy-D-glucose were 8.2- and 4.0-fold higher than those of the wild type, respectively. Y648V had synthetic activity toward N-acetyl-D-glucosamine, while the other variants did not. The oligosaccharide production in the presence of the same concentrations of wild type and each mutant was compared. C485A produced 4-O-beta-D-glucopyranosyl-D-glucosamine from 10 mM Glc1P and D-glucosamine at a rate similar to the wild type. Y648F and Y648V produced 4-O-beta-D-glucopyranosyl-D-mannose and 4-O-beta-D-glucopyranosyl-N-acetyl-D-glucosamine much more rapidly than the wild type when D-mannose and N-acetyl-D-glucosamine were used as acceptors, respectively. After a 4 h reaction, the amounts of 4-O-beta-D-glucopyranosyl-D-mannose and 4-O-beta-D-glucopyranosyl-N-acetyl-D-glucosamine produced by Y648F and Y648V were 5.9- and 12-fold higher than the wild type, respectively. (C) 2013 Elsevier Ltd. All rights reserved.

Structure of a bacterial glycoside hydrolase family 63 enzyme in complex with its glycosynthase product, and insights into the substrate specificity
Takatsugu Miyazaki; Megumi Ichikawa; Gaku Yokoi; Motomitsu Kitaoka; Haruhide Mori; Yoshikazu Kitano; Atsushi Nishikawa; Takashi Tonozuka
FEBS Journal, 280, 4560, 4571, 01 Sep. 2013

Aromatic Residue on beta ->alpha Loop 1 in the Catalytic Domain Is Important to the Transglycosylation Specificity of Glycoside Hydrolase Family 31 alpha-Glucosidase
Kyung-Mo Song; Masayuki Okuyama; Mariko Nishimura; Takayoshi Tagami; Haruhide Mori; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 77, 8, 1759, 1765, Aug. 2013, [Peer-reviewed]
English, Scientific journal

Identification of rice β-glucosidase with high hydrolytic activity towards salicylic acid β-d-glucoside
Nami Himeno; Wataru Saburi; Shinji Wakuta; Ryosuke Takeda; Hideyuki Matsuura; Kensuke Nabeta; Sompong Sansenya; James R; Ketudat Cairns; Haruhide Mori; Ryozo Imai; Hirokazu Matsui
Bioscience, Biotechnology and Biochemistry, 77, 5, 934, 939, 12 Jun. 2013, [Peer-reviewed]
English

Molecular Basis for the Recognition of Long-chain Substrates by Plant alpha-Glucosidases
Takayoshi Tagami; Keitaro Yamashita; Masayuki Okuyama; Haruhide Mori; Min Yao; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 288, 26, 19296, 19303, Jun. 2013, [Peer-reviewed]
English, Scientific journal

Crystal structure of Ruminococcus albus cellobiose 2-epimerase: structural insights into epimerization of unmodified sugar.
Fujiwara T; Saburi W; Inoue S; Mori H; Matsui H; Tanaka I; Yao M
FEBS letters, 587, 7, 840, 846, Apr. 2013, [Peer-reviewed]

Enzymatic Synthesis of Acarviosyl-maltooligosaccharides Using Disproportionating Enzyme 1
Takayoshi Tagami; Yoshiyuki Tanaka; Haruhide Mori; Masayuki Okuyama; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 77, 2, 312, 319, Feb. 2013, [Peer-reviewed]
English, Scientific journal

Identification and characterization of cellobiose 2-epimerases from various aerobes.
Ojima T; Saburi W; Yamamoto T; Mori H; Matsui H
Bioscience, biotechnology, and biochemistry, 77, 1, 189, 193, Taylor & Francis, 2013, [Peer-reviewed]
English, Cellobiose 2-epimerase (CE), found mainly in anaerobes, reversibly converts D-glucose residues at the reducing end of β-1,4-linked oligosaccharides to D-mannose residues. In this study, we characterized CE-like proteins from various aerobes (Flavobacterium johnsoniae NBRC 14942, Pedobacter heparinus NBRC 12017, Dyadobacter fermentans ATCC 700827, Herpetosiphon aurantiacus ATCC 23779, Saccharophagus degradans ATCC 43961, Spirosoma linguale ATCC 33905, and Teredinibacter turnerae ATCC 39867), because aerobes, more easily cultured on a large scale than anaerobes, are applicable in industrial processes. The recombinant CE-like proteins produced in Escherichia coli catalyzed epimerization at the C2 position of cellobiose, lactose, epilactose, and β-1,4-mannobiose, whereas N-acetyl-D-glucosamine, N-acetyl-D-mannosamine, D-glucose, and D-mannose were inert as substrates. All the CEs, except for P. heparinus CE, the optimum pH of which was 6.3, showed highest activity at weakly alkaline pH. CEs from D. fermentans, H. aurantiacus, and S. linguale showed higher optimum temperatures and thermostability than the other enzymes analyzed. The enzymes from D. fermentans, S. linguale, and T. turnerae showed significantly high k_cat and K_m values towards cellobiose and lactose. Especially, T. turnerae CE showed a very high k_cat value towards lactose, an attractive property for the industrial production of epilactose, which is carried out at high substrate concentrations.

A thermophilic alkalophilic α-amylase from Bacillus sp. AAH-31 shows a novel domain organization among glycoside hydrolase family 13 enzymes.
Saburi W; Morimoto N; Mukai A; Kim DH; Takehana T; Koike S; Matsui H; Mori H
Bioscience, biotechnology, and biochemistry, 77, 9, 1867, 1873, Taylor & Francis, 2013, [Peer-reviewed]
English, α-Amylases (EC 3.2.1.1) hydrolyze internal α-1,4-glucosidic linkages of starch and related glucans. Bacillus sp. AAH-31 produces an alkalophilic thermophilic α-amylase (AmyL) of higher molecular mass, 91 kDa, than typical bacterial α-amylases. In this study, the AmyL gene was cloned to determine its primary structure, and the recombinant enzyme, produced in Escherichia coli, was characterized. AmyL shows no hydrolytic activity towards pullulan, but the central region of AmyL (Gly395-Asp684) was similar to neopullulanase-like α-amylases. In contrast to known neopullulanase-like α-amylases, the N-terminal region (Gln29-Phe102) of AmyL was similar to carbohydrate-binding module family 20 (CBM20), which is involved in the binding of enzymes to starch granules. Recombinant AmyL showed more than 95% of its maximum activity in a pH range of 8.2–10.5, and was stable below 65 °C and from pH 6.4 to 11.9. The k_cat values for soluble starch, γ-cyclodextrin, and maltotriose were 103 s⁻¹, 67.6 s⁻¹, and 5.33 s⁻¹, respectively, and the K_m values were 0.100 mg/mL, 0.348 mM, and 2.06 mM, respectively. Recombinant AmyL did not bind to starch granules. But the substitution of Trp45 and Trp84, conserved in site 1 of CBM20, with Ala reduced affinity to soluble starch, while the mutations did not affect affinity for oligosaccharides. Substitution of Trp61, conserved in site 2 of CBM20, with Ala enhanced hydrolytic activity towards soluble starch, indicating that site 2 of AmyL does not contribute to binding to soluble long-chain substrates.

A novel mechanism for the promotion of quercetin glycoside absorption by megalo alpha-1,6-glucosaccharide in the rat small intestine
Aki Shinoki; Weeranuch Lang; Charin Thawornkuno; Hee-Kwon Kang; Yuya Kumagai; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura; Satoshi Ishizuka; Hiroshi Hara
FOOD CHEMISTRY, 136, 2, 293, 296, Jan. 2013, [Peer-reviewed]
English, Scientific journal

Key aromatic residues at subsites +2 and +3 of glycoside hydrolase family 31 alpha-glucosidase contribute to recognition of long-chain substrates
Takayoshi Tagami; Masayuki Okuyama; Hiroyuki Nakai; Young-Min Kim; Haruhide Mori; Kazunori Taguchi; Birte Svensson; Atsuo Kimura
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 1834, 1, 329, 335, Jan. 2013, [Peer-reviewed]
English, Scientific journal

Characterization of a glycoside hydrolase family 31 α-glucosidase involved in starch utilization in podospora anserina
Kyung-Mo Song; Masayuki Okuyama; Kazuyuki Kobayashi; Haruhide Mori; Atsuo Kimura
Bioscience, Biotechnology and Biochemistry, 77, 10, 2117, 2124, 2013, [Peer-reviewed]
English, Scientific journal

Metabolic Mechanism of Mannan in a Ruminal Bacterium, Ruminococcus albus, Involving Two Mannoside Phosphorylases and Cellobiose 2-Epimerase: DISCOVERY OF A NEW CARBOHYDRATE PHOSPHORYLASE, β-1,4-MANNOOLIGOSACCHARIDE PHOSPHORYLASE.
Kawahara R; Saburi W; Odaka R; Taguchi H; Ito S; Mori H; Matsui H
The Journal of biological chemistry, 287, 50, 42389, 42399, 50, Dec. 2012, [Peer-reviewed]
English, Ruminococcus albus is a typical ruminal bacterium digesting cellulose and hemicellulose. Cellobiose 2-epimerase (EC 5.1.3.11, CE), which converts cellobiose to 4-O-β-D-glucosyl-D-mannose, is a particularly unique enzyme in R. albus, but its physiological function is unclear. Recently, a new metabolic pathway of mannan involving CE was postulated for another CE producing bacterium, Bacteroides fragilis. In this pathway, β-1,4-mannobiose is epimerized to 4-O-β-D-mannosyl-D-glucose (Man-Glc) by CE, and Man-Glc is phosphorolyzed to α-D-mannosyl 1-phosphate (Man1P) and D-glucose by Man-Glc phosphorylase (EC 2.4.1.281, MP). Ruminococcus albus NE1 showed intracellular MP activity, and two MP isozymes, RaMP1 and RaMP2, were obtained from the cell-free extract. These enzymes were highly specific for the mannosyl residue at the non-reducing end of the substrate and catalyzed the phosphorolysis and synthesis of Man-Glc through a sequential bi bi mechanism. In a synthetic reaction, RaMP1 showed high activity only towards D-glucose and 6-deoxy-D-glucose in the presence of Man1P, while RaMP2 showed acceptor specificity significantly different from RaMP1. RaMP2 acted on D-glucose derivatives at the C2- and C3-positions including deoxy- and deoxyfluoro-analogues and epimers, but not on those substituted at the C6-position. Furthermore, RaMP2 had high synthetic activity toward the following oligosaccharides: β-linked glucobioses, maltose, N, N'-diacetylchitobiose, and β-1,4-mannooligosaccharides. Particularly, β-1,4-mannooligosaccharides served as significantly better acceptor substrates for RaMP2 than D-glucose. In the phosphorolytic reactions, RaMP2 had weak activity towards β-1,4-mannobiose but efficiently degraded β-1,4-mannooligosaccharides longer than β-1,4-mannobiose. Consequently, RaMP2 is thought to catalyze the phosphorolysis of β-1,4-mannooligosaccharides longer than β-1,4-mannobiose to produce Man1P and β-1,4-mannobiose.

The Delay in the Development of Experimental Colitis from Isomaltosyloligosaccharides in Rats Is Dependent on the Degree of Polymerization
Hitoshi Iwaya; Jae-Sung Lee; Shinya Yamagishi; Aki Shinoki; Weeranuch Lang; Charin Thawornkuno; Hee-Kwon Kang; Yuya Kumagai; Shiho Suzuki; Shinichi Kitamura; Hiroshi Hara; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura; Satoshi Ishizuka
PLOS ONE, 7, 11, 11, e50658, Nov. 2012, [Peer-reviewed]
English, Scientific journal

Amino Acids in Conserved Region II Are Crucial to Substrate Specificity, Reaction Velocity, and Regioselectivity in the Transglucosylation of Honeybee GH-13 alpha-Glucosidases
Lukana Ngiwsara; Gaku Iwai; Takayoshi Tagami; Natsuko Sato; Hiroyuki Nakai; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 76, 10, 1967, 1974, Oct. 2012, [Peer-reviewed]
English, Scientific journal

Bacteroides thetaiotaomicron VPI-5482 glycoside hydrolase family 66 homolog catalyzes dextranolytic and cyclization reactions
Young-Min Kim; Eiji Yamamoto; Min-Sun Kang; Hiroyuki Nakai; Wataru Saburi; Masayuki Okuyama; Haruhide Mori; Kazumi Funane; Mitsuru Momma; Zui Fujimoto; Mikihiko Kobayashi; Doman Kim; Atsuo Kimura
FEBS JOURNAL, 279, 17, 3185, 3191, Sep. 2012, [Peer-reviewed]
English, Scientific journal

Purification and Characterization of a Liquefying alpha-Amylase from Alkalophilic Thermophilic Bacillus sp AAH-31
Dae Hoon Kim; Naoki Morimoto; Wataru Saburi; Atsushi Mukai; Koji Imoto; Toshihiko Takehana; Seiji Koike; Haruhide Mori; Hirokazu Matsui
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 76, 7, 1378, 1383, Jul. 2012
English, Scientific journal

Structural Elucidation of Dextran Degradation Mechanism by Streptococcus mutans Dextranase Belonging to Glycoside Hydrolase Family 66
Nobuhiro Suzuki; Young-Min Kim; Zui Fujimoto; Mitsuru Momma; Masayuki Okuyama; Haruhide Mori; Kazumi Funane; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 287, 24, 19916, 19926, Jun. 2012, [Peer-reviewed]
English, Scientific journal

Novel Dextranase Catalyzing Cycloisomaltooligosaccharide Formation and Identification of Catalytic Amino Acids and Their Functions Using Chemical Rescue Approach
Young-Min Kim; Yoshiaki Kiso; Tomoe Muraki; Min-Sun Kang; Hiroyuki Nakai; Wataru Saburi; Weeranuch Lang; Hee-Kwon Kang; Masayuki Okuyama; Haruhide Mori; Ryuichiro Suzuki; Kazumi Funane; Nobuhiro Suzuki; Mitsuru Momma; Zui Fujimoto; Tetsuya Oguma; Mikihiko Kobayashi; Doman Kim; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 287, 24, 19927, 19935, Jun. 2012, [Peer-reviewed]
English, Scientific journal

A Novel Metabolic Pathway for Glucose Production Mediated by alpha-Glucosidase-catalyzed Conversion of 1,5-Anhydrofructose
Young-Min Kim; Wataru Saburi; Shukun Yu; Hiroyuki Nakai; Janjira Maneesan; Min-Sun Kang; Seiya Chiba; Doman Kim; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 287, 27, 22441, 22444, Jun. 2012, [Peer-reviewed]
English, Scientific journal

Enzymatic Characteristics of Cellobiose Phosphorylase from Ruminococcus albus NE1 and Kinetic Mechanism of Unusual Substrate Inhibition in Reverse Phosphorolysis
Ken Hamura; Wataru Saburi; Shotaro Abe; Naoki Morimoto; Hidenori Taguchi; Haruhide Mori; Hirokazu Matsui
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 76, 4, 812, 818, Apr. 2012
English, Scientific journal

Chemical constituents and free radical scavenging activity of corn pollen collected from Apis mellifera hives compared to floral corn pollen at Nan, Thailand
Atip Chantarudee; Preecha Phuwapraisirisan; Kiyoshi Kimura; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura; Chanpen Chanchao
BMC COMPLEMENTARY AND ALTERNATIVE MEDICINE, 12, 45, Apr. 2012, [Peer-reviewed]
English, Scientific journal

In vitro antiproliferative/cytotoxic activity on cancer cell lines of a cardanol and a cardol enriched from Thai Apis mellifera propolis
Dungporn Teerasripreecha; Preecha Phuwapraisirisan; Songchan Puthong; Kiyoshi Kimura; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura; Chanpen Chanchao
BMC COMPLEMENTARY AND ALTERNATIVE MEDICINE, 12, 27, Mar. 2012, [Peer-reviewed]
English, Scientific journal

Immobilization of a thermostable cellobiose 2-epimerase from Rhodothermus marinus JCM9785 and continuous production of epilactose.
Sato H; Saburi W; Ojima T; Taguchi H; Mori H; Matsui H
Bioscience, biotechnology, and biochemistry, 76, 8, 1584, 1587, 8, 2012, [Peer-reviewed]
English, Cellobiose 2-epimerase (CE) efficiently forms epilactose which has several beneficial biological functions. A thermostable CE from Rhodothermus marinus was immobilized on Duolite A568 and packed into a column. Lactose (100 g/L) was supplied to the reactor, kept at 50 °C at a space velocity of 8 h⁻¹. The epilactose concentration of the resulting eluate was 30 g/L, and this was maintained for 13 d.

Degree of polymerization in dietary α-1,6-gluco¬sac¬cha¬rides modulates symptom of experimental colitis in rats.
Iwaya H; Lee JS; Yamagishi S; Shinoki A; Lang W; Kang HK; Okuyama M; Mori H; Hara H; Kimura A; Ishizuka S
Plos One, 7, 11, e50658, e50658, 2012, [Peer-reviewed]
English, Scientific journal

Characterization of some enzymatic properties of recombinant α-glucosidase III from the Thai honeybee, Apis cerana indica Fabricus.
Kaewmuangmoon J; Yoshiyama M; Kimura K; Okuyama M; Mori H; Kimura A; Chanchao C
Afr J Biotechnol, 11, 96, 16220, 16232, 2012, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary crystallographic analysis of dextranase from Streptococcus mutans
Nobuhiro Suzuki; Young-Min Kim; Zui Fujimoto; Mitsuru Momma; Hee-Kwon Kang; Kazumi Funane; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 67, 12, 1542, 1544, Dec. 2011
English, Scientific journal

Biochemical Characterization of a Thermophilic Cellobiose 2-Epimerase from a Thermohalophilic Bacterium, Rhodothermus marinus JCM9785
Teruyo Ojima; Wataru Saburi; Hiroki Sato; Takeshi Yamamoto; Haruhide Mori; Hirokazu Matsui
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 75, 11, 2162, 2168, Nov. 2011
English, Scientific journal

Transglycosylation by barley alpha-amylase 1
Janos A. Motyan; Erika Fazekas; Haruhide Mori; Birte Svensson; Peter Bagossi; Lili Kandra; Gyoengyi Gyemant
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 72, 3-4, 229, 237, Nov. 2011, [Peer-reviewed]
English, Scientific journal

Calcium Ion-Dependent Increase in Thermostability of Dextran Glucosidase from Streptococcus mutans
Momoko Kobayashi; Hironori Hondoh; Haruhide Mori; Wataru Saburi; Masayuki Okuyama; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 75, 8, 1557, 1563, Aug. 2011
English, Scientific journal

Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity
Young-Min Kim; Ryoko Shimizu; Hiroyuki Nakai; Haruhide Mori; Masayuki Okuyama; Min-Sun Kang; Zui Fujimoto; Kazumi Funane; Doman Kim; Atsuo Kimura
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 91, 2, 329, 339, Jul. 2011
English, Scientific journal

Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity
Young-Min Kim; Ryoko Shimizu; Hiroyuki Nakai; Haruhide Mori; Masayuki Okuyama; Min-Sun Kang; Zui Fujimoto; Kazumi Funane; Doman Kim; Atsuo Kimura
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 91, 2, 329, 339, Jul. 2011, [Peer-reviewed]
English, Scientific journal

Comparison of Enzymatic Properties and Gene Expression Profiles of Two Tuberonic Acid Glucoside β-Glucosidases from Oryza sativa L.
WAKUTA Shinji; HAMADA Shigeki; ITO Hiroyuki; IMAI Ryozo; MORI Haruhide; MATSUURA Hideyuki; NABETA Kensuke; MATSUI Hirokazu
Journal of applied glycoscience, 58, 2, 67, 70, Japanese Society of Applied Glycoscience, 20 Apr. 2011
English

Suicide Substrate-based Inactivation of Endodextranase by ω-Epoxyalkyl α-D-Glucopyranosides
KANG Hee-Kwon; KIM Young-Min; NAKAI Hiroyuki; KANG Min-Sun; HAKAMADA Wataru; OKUYAMA Masayuki; MORI Haruhide; NISHIO Toshiyuki; KIMURA Atsuo
Journal of Applied Glycoscience, 57, 4, 269, 272, The Japanese Society of Applied Glycoscience, 20 Oct. 2010
English, Three kinds of ω-epoxyalkyl α-glucopyranosides (3′,4′-epoxybutyl α-D-glucopyranoside (E4G), 4′,5′-epoxypentyl α-D-glucopyranoside (E5G) and 5′,6′-epoxyhexyl α-D-glucopyranoside (E6G)), having alkyl chains of different lengths at their aglycone moieties, inactivated the endodextranase from Streptococcus mutans ATCC 25175 (SmDex) irreversibly with the pseudo-first order kinetics. Alkyl chain length-dependent inactivation was observed and the degree of activity loss was E5G, E6G and E4G, in that order, implying that the distance between epoxide group and glucosyl residue of ω-epoxyalkyl α-glucopyranoside was important in the modification of endodextranase. Inactivation by E5G followed the model of reversible intermediate-complex formation mechanism (suicide inhibitor-based mechanism). The rate constant of irreversible inactivation (k) and the dissociation constant of intermediate-complex (KR) of SmDex and E5G were 0.44 min^-1 and 1.45 mM, respectively. Hydrolytic reaction product (isomaltose) protected SmDex from E5G-inactivation, suggesting that E5G bound to the catalytic site of SmDex. This is the first report that ω-epoxyalkyl α-glucopyranoside becomes a suicide substrate for endodextranase.

Suicide Substrate-based Inactivation of Endodextranase by .OMEGA.-Epoxyalkyl .ALPHA.-D-Glucopyranosides
カンヒゴン,キムヨンミン; 中井博之; カンミンソン; 袴田航; 奥山正幸; 森春英; 西尾俊幸; 木村淳夫
J Appl Glycosci, 57, 4, 269-272 (J-STAGE), 272, The Japanese Society of Applied Glycoscience, 2010
English, Three kinds of ω-epoxyalkyl α-glucopyranosides (3′,4′-epoxybutyl α-D-glucopyranoside (E4G), 4′,5′-epoxypentyl α-D-glucopyranoside (E5G) and 5′,6′-epoxyhexyl α-D-glucopyranoside (E6G)), having alkyl chains of different lengths at their aglycone moieties, inactivated the endodextranase from Streptococcus mutans ATCC 25175 (SmDex) irreversibly with the pseudo-first order kinetics. Alkyl chain length-dependent inactivation was observed and the degree of activity loss was E5G, E6G and E4G, in that order, implying that the distance between epoxide group and glucosyl residue of ω-epoxyalkyl α-glucopyranoside was important in the modification of endodextranase. Inactivation by E5G followed the model of reversible intermediate-complex formation mechanism (suicide inhibitor-based mechanism). The rate constant of irreversible inactivation (k) and the dissociation constant of intermediate-complex (KR) of SmDex and E5G were 0.44 min^-1 and 1.45 mM, respectively. Hydrolytic reaction product (isomaltose) protected SmDex from E5G-inactivation, suggesting that E5G bound to the catalytic site of SmDex. This is the first report that ω-epoxyalkyl α-glucopyranoside becomes a suicide substrate for endodextranase.

Function analysis of subsite+1 mutants of isomaltooligosaccharide 6-α-glucosyltransferase
nishimura takashi; kanegae michiyo; hondoh hironori; okuyama masayuki; mori haruhide; kimura atsuo
Journal of Applied Glycoscience Supplement, 2010, 71, 71, The Japanese Society of Applied Glycoscience, 2010
Japanese

Modification of subsite +1 in α-glucosidase derived from Aspergillus niger
Tagami Takayoshi; Okuyama Masayuki; Mori Haruhide; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2010, 73, 73, The Japanese Society of Applied Glycoscience, 2010
Japanese

Catalytic Reaction Mechanism Based on alpha-Secondary Deuterium Isotope Effects in Hydrolysis of Trehalose by European Honeybee Trehalase
Haruhide Mori; Jin-Ha Lee; Masayuki Okuyama; Mamoru Nishimoto; Masao Ohguchi; Doman Kim; Atsuo Kimura; Seiya Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 73, 11, 2466, 2473, Nov. 2009
English, Scientific journal

The first alpha-1,3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31
Min-Sun Kang; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
BIOCHIMIE, 91, 11-12, 1434, 1442, Nov. 2009, [Peer-reviewed]
English, Scientific journal

Catalytic Mechanism of Retaining alpha-Galactosidase Belonging to Glycoside Hydrolase Family 97
Masayuki Okuyama; Momoyo Kitamura; Hironori Hondoh; Min-Sun Kang; Haruhide Mori; Atsuo Kimura; Isao Tanaka; Min Yao
JOURNAL OF MOLECULAR BIOLOGY, 392, 5, 1232, 1241, Oct. 2009, [Peer-reviewed]
English, Scientific journal

Role of Ca2+ in catalysis of SusB derived from Bacteroides thetaiotaomicron
Yoshida Takuya; Okuyama Masayuki; Hondoh Hironori; Yao Min; Mori Haruhide; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2009, 38, 38, The Japanese Society of Applied Glycoscience, 2009
Japanese

Identification of a key residue for forming of subsites +2 and +3 in sugar beet alpha;-glucosidase
Tagami Takayoshi; Okuyama Masayuki; Mori Haruhide; Taguchi Kazunori; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2009, 37, 37, The Japanese Society of Applied Glycoscience, 2009
Japanese

Alteration of transglucosylation/hydrolysis ratio of Streptococcus mutans dextran glucosidase (2)
Nakatsuka Daichi; Hondoh Hironori; Otsuka Hiroaki; Saburi Wataru; Mori Haruhide; Okuyama Masayuki; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2009, 34, 34, The Japanese Society of Applied Glycoscience, 2009
Japanese

Improving Activity of Honeybee α-Glucosidase III by Substitution of Q349 and L350 Position.
Ngiwsara Lukana; Mori Haruhide; Okuyama Masayuki; Chiba Seiya; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2009, 36, 36, The Japanese Society of Applied Glycoscience, 2009
Japanese

Structural element for transglucosylation reaction of isomaltooligosaccharide 6-α-glucosyltransferase
Nishimura Takashi; Kanegae Michiyo; KIM Young-Min; Hondoh Hironori; Okuyama Masayuki; Mori Haruhide; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2009, 35, 35, The Japanese Society of Applied Glycoscience, 2009
Japanese

Structure-function relationship of substrate length specificity of dextran glucosidase from Streptococcus mutans
Wataru Saburi; Hironori Hondoh; Young-Min Kim; Haruhide Mori; Masayuki Okuyama; Atsuo Kimura
BIOLOGIA, 63, 6, 1000, 1005, Dec. 2008
English, Scientific journal

Structural and Functional Analysis of a Glycoside Hydrolase Family 97 Enzyme from Bacteroides thetaiotaomicron
Momoyo Kitamura; Masayuki Okuyama; Fumiko Tanzawa; Haruhide Mori; Yu Kitago; Nobuhisa Watanabe; Atsuo Kimura; Isao Tanaka; Min Yao
JOURNAL OF BIOLOGICAL CHEMISTRY, 283, 52, 36328, 36337, Dec. 2008, [Peer-reviewed]
English, Scientific journal

Molecular Mechanism of α-glucosidase
Masayuki Okuyama; Haruhide Mori; Hironori Hondoh; Hiroyuki Nakai; Wataru Saburi; Min Sung Kang; Young Min Kim; Mamoru Nishimoto; Jintanart Wongchawalit; Takeshi Yamamoto; Mee Son; Jin Ha Lee; San San Mar; Kenji Fukuda; Seiya Chiba; Atsuo Kimura
Carbohydrate-Active Enzymes: Structure, Function and Applications, 64, 76, Sep. 2008, [Peer-reviewed]
In book

Rice α-glucosidase isozymes and isoforms showing different starch granules-binding and -degrading ability
Nakai H; Tanizawa S; Ito T; Kamiya K; Kim YM; Yamamoto T; Matsubara K; Sakai M; Sato H; Imbe T; Okuyama M; Mori H; Chiba S; Sano Y; Kimura A
Biocatalysis and Biotransformation, 26, 104, 110, Jul. 2008, [Peer-reviewed]
English, Scientific journal

Substrate Recognition of Escherichia coli YicI (.ALPHA.-Xylosidase)
奥山正幸; カンミンソン; 矢追克郎; 三石安; 森春英; 木村淳夫
J Appl Glycosci, 55, 2, 111-118 (J-STAGE), 118, The Japanese Society of Applied Glycoscience, 2008
English, Glycoside hydrolase family 31 (GH 31) is one of the most intriguing glycoside hydrolase families. This family contains α-glucosidase, α-xylosidase, α-glucan lyase and isomaltosyltransferase. Escherichia coli YicI (α-xylosidase) is a representative enzyme of GH 31 because its biochemical and structural studies have been thoroughly carried out. YicI is a strict α-xylosidase, which rigidly recognizes α-xyloside at the non-reducing terminal end, even though its amino acid sequence apparently displays similarity with α-glucosidases. Phe277, Cys307, Trp345 and Lys414 at the subsite-1 are important for α-xylosidase activity. The mutant YicI enzymes, which possesses Ile307/Asp308 instead of Cys307/Phe308 and which has a shorter β→α loop 1 of (β/α)8 barrel in place of the original longer loop, respectively, possess α-glucosidase activity. In the transxylosylation of YicI, glucose, mannose and allose are able to act as acceptors, but galactose, talose and gulose never do, implying that equatorial OH-4 of the aldopyranose is crucial for acting as an acceptor. YicI transfers α-xylosyl moieties to a specific hydroxy group in the acceptor sugar (except fructopyranose) showing 1,6 regioselectivity, which is in agreement with the structural feature of the aglycone-biding site. Among the transxylosylation products of YicI, α-D-xylopyranosyl-(1→6)-D-mannopyranose, α-D-xylopyranosyl-(1→6)-D-fructofuranose, and α-D-xylopyranosyl-(1→3)-D-fructopyranose are novel sugars. α-D-Xylopyranosyl-(1→6)-D-mannopyranose and α-D-xylopyranosyl-(1→6)-D-fructofuranose have the ability to inhibit rat intestinal α-glucosidases.

The structural factor for transglucosylation of isomaltooligosaccharide 6-α-glucosyltrasferase(I6GT) from Bacillus sp.
Kanegae Michiyo; KIM Young-Min; Hondoh Hironori; Okuyama Masayuki; Mori Haruhide; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2008, 104, 104, The Japanese Society of Applied Glycoscience, 2008
Japanese

Catalytic mechanism of retaining glycosidase in glycoside hydrolase family 97
OKUYAMA MASAYUKI; YAO MIN; Hondoh Hironori; KITAMURA MOMOYO; MORI HARUHIDE; TANAKA ISAO; KIMURA ATSUO
Journal of Applied Glycoscience Supplement, 2008, 139, 139, The Japanese Society of Applied Glycoscience, 2008
Japanese

Cloning and Expression of α-glucosidase derived from ripening seed of Sugar Beet
Tagami Takayoshi; Okuyama Masayuki; Mori Haruhide; Taguchi Kazunori; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2008, 94, 94, The Japanese Society of Applied Glycoscience, 2008
Japanese

Coversion of Lactobacillus johnsonii novel α-glucosidase into glycosynthase
Kang Min-Sun; Okuyama Masayuki; Mori Haruhide; Kimura Atsuo
Journal of Applied Glycoscience Supplement, 2008, 96, 96, The Japanese Society of Applied Glycoscience, 2008
Japanese

Glycoside hydrolase family 31 Escherichia coli alpha-xylosidase
M-S. Kang; M. Okuyama; K. Yaoi; Y. Mitsuishi; Y-M. Kim; H. Mori; A. Kimura
BIOCATALYSIS AND BIOTRANSFORMATION, 26, 1-2, 96, 103, 2008
English, Scientific journal

Substrate recognition mechanism of alpha-1,6-glucosidic linkage hydrolyzing enzyme, dextran glucosidase from Streptococcus mutans.
Hondoh H; Saburi W; Mori H; Okuyama M; Nakada T; Matsuura Y; Kimura A
J Mol Biol, 378, 4, 913, 922, 2008, [Peer-reviewed]

Function-unknown glycoside hydrolase family 31 proteins, mRNAs of which were expressed in rice ripening and germinating stages, are alpha-glucosidase and alpha-xylosidase
Hiroyuki Nakai; Shigeki Tanizawa; Tatsuya Ito; Koutarou Kamiya; Young-Min Kim; Takeshi Yamamoto; Kazuki Matsubara; Makoto Sakai; Hiroyuki Sato; Tokio Imbe; Masayuki Okuyama; Haruhide Mori; Yoshio Sano; Seiya Chiba; Atsuo Kimura
JOURNAL OF BIOCHEMISTRY, 142, 4, 491, 500, Oct. 2007
English, Scientific journal

Crystallization and preliminary X-ray analysis of Streptococcus mutans dextran glucosidase
Wataru Saburi; Hironori Hondoh; Hideaki Unno; Masayuki Okuyama; Haruhide Mori; Toshitaka Nakada; Yoshiki Matsuura; Atsuo Kimura
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 63, 9, 774, 776, Sep. 2007
English, Scientific journal

Molecular cloning of cDNA for trehalase from the European honeybee, Apis mellifera L., and its heterologous expression in Pichia pastoris
Jin-Ha Lee; Saori Saito; Haruhide Mori; Mamoru Nishimoto; Masayuki Okuyama; Doman Kim; Jintanart Wongchawalit; Atsuo Kimura; Seiya Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 9, 2256, 2265, Sep. 2007
English, Scientific journal

Molecular cloning of cDNAs and genes for three alpha-glucosidases from European honeybees, Apis mellifera L., and heterologous production of recombinant enzymes in Pichia pastoris
Mamoru Nishimoto; Haruhide Mori; Tsuneharu Moteki; Yukiko Takamura; Gaku Iwai; Yu Miyaguchi; Masayuki Okuyama; Jintanart Wongchawalit; Rudee Surarit; Jisnuson Svasti; Atsuo Kimura; Seiya Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 7, 1703, 1716, Jul. 2007
English, Scientific journal

Multiple forms of alpha-glucosidase in rice seeds (Oryza sativa L., var Nipponbare)
Hiroyuki Nakai; Tatsuya Ito; Masatoshi Hayashi; Koutarou Kamiya; Takeshi Yamamoto; Kazuki Matsubara; Young-Min Kim; Wongchawalit Jintanart; Masayuki Okuyama; Haruhide Mori; Seiya Chiba; Yoshio Sano; Atsuo Kimura
BIOCHIMIE, 89, 1, 49, 62, Jan. 2007
English, Scientific journal

Purification and characterization of alpha-glucosidase I from Japanese honeybee (Apis cerana japonica) and molecular cloning of its cDNA
Jintanart Wongchawalit; Takeshi Yamamoto; Hiroyuki Nakai; Young-Min Kim; Natsuko Sato; Mamoru Nishimoto; Masayuki Okuyama; Haruhide Mori; Osamu Saji; Chanpen Chanchao; Siriwat Wongsiri; Rudee Surarit; Jisnuson Svasti; Seiya Chiba; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70, 12, 2889, 2898, Dec. 2006
English, Scientific journal

Interactions between Barley α-Amylases, Substrates, Inhibitors and Regulatory Proteins
HACHEM Maher Abou; BOZONNET Sophie; WILLEMOES Martin; BONSAGER Birgit C; NIELSEN Morten Munch; FUKUDA Kenji; KRAMHOFT Birte; MAEDA Kenji; SIGURSKJOLD Bent W; HAGGLUND Per; FINNIE Christine; MORI Haruhide; ROBERT Xavier; JENSEN Malene H; TRANIER Samuel; AGHAJARI Nushin; HASER Richard; SVENSSON Birte
Journal of applied glycoscience, 53, 2, 163, 169, The Japanese Society of Applied Glycoscience, 20 Apr. 2006
English, Barley α-amylase binds sugars at two sites on the enzyme surface in addition to the active site. Crystallography and site-directed mutagenesis highlight the importance of aromatic residues at these surface sites as demonstrated by K_d values determined for β-cyclodextrin by surface plasmon resonance and for starch granules by adsorption analysis. Activity towards amylopectin and amylose follows two different kinetic models, degradation of amylopectin being composed of a fast and a slow component, perhaps reflecting attack on A and B chains, respectively, whereas amylose hydrolysis follows a simple Michaelian kinetics. β-cyclodextrin binding at surface sites inhibits only the fast reaction in amylopectin degradation. Site-directed mutagenesis and activity analysis, furthermore show that one of the surface binding sites as well as individual subsites in the active site cleft have distinct roles in the multiple attack on amylose. Although the two isozymes AMY1 and AMY2 share ligands for three structural calcium ions, they differ importantly in the effect of calcium on activity and stability, AMY1 having the higher affinity and the lower stability. The role of the individual calcium ions is studied by mutagenesis, crystallography and microcalorimetry. Further improvement of recombinant AMY2 production allows future direct mutational analysis in this isozyme. Specific proteinaceous inhibitors act on α-amylases of different origin. In the complex of barley α-amylase/subtilisin inhibitor (BASI) with AMY2, a fully hydrated calcium ion at the protein interface mediates contact between inhibitor residues and the enzyme catalytic groups in a manner that depends on calcium and which can be suppressed by site-directed mutagenesis of Glu168 in BASI. Finally certain inhibitors and enzymes are targets of the disulphide reductase thioredoxin h that attacks a specific disulphide bond in BASI and, remarkably, reduces two different disulphide bonds in the barley monomeric and dimeric amylase inhibitors that both belong to the CM-proteins and inhibit animal α-amylase.

Plant α-Glucosidase : Molecular Analysis of Rice α-Glucosidase and Degradation Mechanism of Starch Granules in Germination Stage
NAKAI Hiroyuki; ITO Tatsuya; TANIZAWA Shigeki; MATSUBARA Kazuki; YAMAMOTO Takeshi; OKUYAMA Masayuki; MORI Haruhide; CHIBA Seiya; SANO Yoshio; KIMURA Atsuo
Journal of applied glycoscience, 53, 2, 137, 142, The Japanese Society of Applied Glycoscience, 20 Apr. 2006
English, In germination of plant seeds, storage starch is principally degraded by the combination of amylolytic enzymes. As starch is an insoluble granule, a conventional view of the degradation pathway is that the initial attack is performed by α-amylase having the starch granule-binding ability. Plant α-glucosidase was also capable of adsorbing and hydrolyzing starch granules directly, indicating a possible second pathway: the direct liberation of glucose from starch granules by plant α-glucosidase rather than the α-amylase-mediated system. We found that the starch-binding site of plant α-glucosidase was situated in its C-terminal region, of which function was independent of the catalytic domain. Site-directed mutagenesis analysis on the aromatic amino acid residues conserved in this region revealed that Trp803 and Phe895 of rice α-glucosidase were responsible for binding to starch granules. Mold α-glucosidases were devoid of the ability to attack starch granules. In plant seeds, multiple α-glucosidases have been observed. Two types of α-glucosidases, insoluble and soluble enzymes, were found in the germinating stage of rice. Expression patterns of their activities classified 14 rice varieties into two groups (Groups 1 and 2). In Group 1 varieties, insoluble enzyme decreased immediately after germination. The soluble enzyme increased by de novo synthesis. Group 2 maintained a constant activity level of insoluble and soluble α-glucosidases in germination. From Groups 1 and 2, we selected varieties of Akamai and Nipponbare, respectively, of which analysis elucidated interesting molecular mechanisms of insoluble and soluble enzymes: i) isoform and isozyme formations by post-translational proteolysis as well as by chromosomal gene expression; ii) characterization of purified enzymes exhibiting different activities to starch granules.

Identification and Manipulation of Subsite Structure and Starch Granule Binding Site in Plant .ALPHA.-Amylase
MORI HARUHIDE
J Appl Glycosci, 53, 1, 51, 56, 20 Jan. 2006
Japanese

Developmental regulation of photosynthate distribution in leaves of rice
T Shinano; K Nakajima; J Wasaki; H Mori; T Zheng; M Osaki
PHOTOSYNTHETICA, 44, 1, 1, 10, 2006
English, Scientific journal

Glucoamylase originating from Schwanniomyces occidentalis is a typical alpha-glucosidase
F Sato; M Okuyama; H Nakai; H Mori; A Kimura; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 69, 10, 1905, 1913, Oct. 2005
English, Scientific journal

Oligosaccharide binding to barley alpha-amylase 1
Robert, X; R Haser; H Mori; B Svensson; N Aghajari
JOURNAL OF BIOLOGICAL CHEMISTRY, 280, 38, 32968, 32978, Sep. 2005, [Peer-reviewed]
English, Scientific journal

Binding of carbohydrates and protein inhibitors to the surface of α-amylases
Sophie Bozonnet; Birgit C. Boønsager; Birte Kramhøft; Haruhide Mori; Maher Abou Hachem; Martin Willemoës; Morten T. Jensen; Kenji Fukuda; Peter K. Nielsen; Nathalie Juge; Nushin Aghajari; Samuel Tranier; Xavier Robert; Richard Haser; Birte Svensson
Biologia - Section Cellular and Molecular Biology, 60, SUPPL. 16, 27, 36, 2005, [Peer-reviewed]
International conference proceedings

Molecular analysis of α-glucosidase belonging to GH-family 31
Nakai H; Okuyama M; Kim YM; Saburi W; Wongchawalit J; Mori H; Chiba S; Kimura A
Biologia, Bratislava, 60, 131, 135, 2005, [Peer-reviewed]
English

Localization of alpha-glucosidases I, II, and III in organs of European honeybees, Apis mellifera L., and the origin of alpha-glucosidase in honey
M Kubota; M Tsuji; M Nishimoto; J Wongchawalit; M Okuyama; H Mori; H Matsui; R Surarit; J Svasti; A Kimura; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68, 11, 2346, 2352, Nov. 2004
English, Scientific journal

An improved method for deleting large regions of Escherichia coli K-12 chromosome using a combination of Cre/loxP and lambda Red.
Fukiya S; Mizoguchi H; Mori H
FEMS microbiology letters, 234, 2, 325, 331, 2, May 2004, [Peer-reviewed]

Barley Proteome Analysis, Starch Degrading Eznymes and Proteinaceous Inhibitors
FINNIE Christine; OSTERGAARD Ole; BAK-JENSEN Kristian Sass; NIELSEN Peter K; BONSAGER Birgit C; MORI Haruhide; NOHR Jane; KRAMHOFT Birte; JUGE Nathalie; SVENSSON Birte
Journal of Applied Glycoscience, 50, 2, 277, 282, The Japanese Society of Applied Glycoscience, 14 Jul. 2003
English, Proteomes of barley seeds were described by 2-D gel electrophoresis and spots selected for proteinidentification by mass spectrometry and database searches. Proteins were categorised according to temporalappearance during seed development and maturation. Fragments of β-amylases appeared transiently at midgrain filling and during germination. The α-amylase/trypsin inhibitors increased during grain filling andtypical housekeeping enzymes were present throughout the period. Germination altered the proteome and dissection of micromalted seeds enabled localization of selected proteins to specifi...

Impact on Substrate Specificity of Mutational Subsite Isozyme Mimicry in Barley α-Amylase
SVENSSON Birte; MORI Haruhide; BAK-JENSEN Kristian Sass; JENSEN Morten Tovborg
Journal of Applied Glycoscience, 50, 2, 143, 145, The Japanese Society of Applied Glycoscience, 14 Jul. 2003
English, The mutational analysis of the roles of specific side chains at individual subsites have been conducted for barley α-amylase 1(AMY1) across the ten subsites long substrate binding cleft. The present study specifically focuses on such mutants in which the AMY2 structure has been mimicked. Generally the kinetics parameters for mutants at subsites accommodating the substrate glycone part showed decreased affinity for oligosaccharide and amylose DP 17 whereas an aglycon binding subsite +4 AMY2 mimic had increased affinity but reduced activity. Among barley α-amylase/subtilisin inhibitor (BASI) ...

Purification, characterization, and sequence analysis of two alpha-amylase isoforms from azuki bean, Vigna angularis, showing different affinity towards beta-cyclodextrin sepharose
SS Mar; H Mori; JH Lee; K Fukuda; W Saburi; A Fukuhara; M Okuyama; S Chiba; A Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 67, 5, 1080, 1093, May 2003
English, Scientific journal

Evidence of Intramolecular Transglucosylation Cathlyzed by an α-Glucosidase
SON Mee; MORI Haruhide; OKUYAMA Masayuki; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 50, 1, 41, 44, The Japanese Society of Applied Glycoscience, 20 Jan. 2003
English, The hydrolytic reaction of carbohydrate-hydrolase is essentially accompanied by a reverse reaction (the condensation reaction), meaning that only the substrate capable of being hydrolyzed is produced by the reverse reaction. Honeybee α-glucosidase I can't hydrolyze isomaltose, but is capable of hydrolyzing maltose, kojibiose and slightly nigerose. Nevertheless, the enzyme catalyzes the formation and accumulation of isomaltose from glucose together with α-glucobioses such as maltose, kojibiose and nigerose. This finding is in conflict with the data that the enzyme has no hydrolytic activity toward isomaltose. However, the conflict for the peculiar phenomenon on the reaction was rationally explained by the evidence that isomaltose might be formed by the intramolecular transglucosylation via other α-glucobioses that are easily produced from glucose by the condensation reaction. It is suggested that the usual transglycosylation of carbohydrate-hydrolase may be accompanied by an intramolecular transfer reaction.

Evidence of Intramolecular Transglucosylation Catalyzed by an .ALPHA.-Glucosidase.
Son Mee; Mori Haruhide; Okuyama Masayuki; Kimura Atsuo; Chiba Seiya
Journal of Applied Glycoscience, 50, 1, 41, 44, 日本応用糖質科学会, 2003
English, The hydrolytic reaction of carbohydrate-hydrolase is essentially accompanied by a reverse reaction (the condensation reaction), meaning that only the substrate capable of being hydrolyzed is produced by the reverse reaction. Honeybee α-glucosidase I can't hydrolyze isomaltose, but is capable of hydrolyzing maltose, kojibiose and slightly nigerose. Nevertheless, the enzyme catalyzes the formation and accumulation of isomaltose from glucose together with α-glucobioses such as maltose, kojibiose and nigerose. This finding is in conflict with the data that the enzyme has no hydrolytic activity ...

Identification of essential ionizable groups and evaluation of subsite affinities in the active site of beta-D-glucosidase F-1 from a Streptomyces sp.
K Fukuda; H Mori; M Okuyama; A Kimura; H Ozaki; M Yoneyama; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66, 10, 2060, 2067, Oct. 2002
English, Scientific journal

Kinetic Studies on Substrate Specificity and Active Site of β-_D-Glucosidase F_1 from Streptomyces sp.
FUKUDA Kenji; SHIRAKAWA Kou; MORI Haruhide; OKUYAMA Masayuki; KIMURA Atsuo; OZAKI Hachiro; YONEYAMA Michio; CHIBA Seiya
Journal of applied glycoscience, 49, 3, 265, 272, The Japanese Society of Applied Glycoscience, 18 Jul. 2002
English, The substrate specificity and the active site of β-D-glucosidase F^-1 (Mr, 50, 000; optimum pH, 5.5) purified from a Streptomyces sp. were kinetically investigated. The β-D-glucosidase showed a broad substrate specificity for synthetic glycosides and disaccharides having β-glycosidic linkage, but the former was more favorable substrate than the latter. The enzyme was characterized by the ability to hydrolyze rapidly not only ρ-nitrophenyl β-glucoside (K_m, 0.72 mM; k₀, 63 s^-1) but also ρ-nitrophenyl β-fucoside (K_m, 0.19 mM; k₀, 44 s^-1) and laminaribiose (K_m, 1.6 mM; k₀, 70 s^-1). The kinetic study was made as to whether the hydrolyses of these synthetic glycosides were catalyzed at a single active site or at dual active sites in the β-D-glucosidase. In the experiments with the mixed substrates of p-nitrophenyl β-glucoside (PNPG) and ρ-nitrophenyl β-fucoside (PNPF) or ρ-nitrophenyl β-galactoside (PNPGal), the kinetic features, the linearity of Lineweaver-Burk plots and the dependence of the apparent maximal veloities and K_m value on the mole fraction (f) of PNPG in the mixed substrate, f = [PNPG/([PNPG] + [PNPF or PNPGal]) agreed very closely with those theoretically predicted for a single catalytic site mechanism. The findings strongly support the notion that the β-D-glucosidase attacks the synthetic glycosides at a common active site.

Study on Three .ALPHA.-Glucosidase Isozymes from Honeybee, Apis mellifera L.
NISHIMOTO MAMORU; MORI HARUHIDE; KIMURA ATSUO; CHIBA SEIYA
J Appl Glycosci, 49, 2, 191, 197, The Japanese Society of Applied Glycoscience, 01 Apr. 2002
Japanese, The genes of three a-glucosidases (HBG I, HBG II and HBG III) were isolated from the cDNA library of honeybee, Apis mellifera L. The nucleotide sequences of HBG I, II and III were consisted of 1974, 1910 and 1916 base pair and encoding 588, 580 and 567 amino acid residues, respectively. The putative primary structures showed high homology ranging from N- to C-terminals, and three enzymes belonged to a-glucosidase family I, in which four conservative regions of aamylase family were observed in their sequences. To obtain the recombinant enzymes, we tried to express the cDNAs in Pichia pastoris of heterologous host cells. Although recombinant HBG I was not produced, the recombinant HBG II and III of 2.4 and 1.2 U/mg were respectively expressed and secreted into culture supernatant. The active recombinant enzymes purified had the same properties as those of native ones except sugar content. To investigate the catalytic residues in HBGs, four mutated enzymes (D206N, E259Q, E269Q and D33 1N) of HBG III were constructed, and their specific activities were found to be 0.0004, 4.9, 0.004 and 0.0002 U/mg, respectively. E259Q remained half activity of wild type and those of the others disappeared, implying that three catalyticresidues of HBGs were D212, E281 and D343 of HBG I, D202, E271 and D333 of HBG II, D206, E269 and D331 of HBG III. The homology modeling showed that three enzymes had Ndomain ((β/α)₈ barrel), subdomain, and C-domain(β-sheet structure mainly) like oligo-l, 6-glucosidase from Bacillus cereus.

Catalytic Amino Acid Residue Providing Proton Donor in .ALPHA.-Glucosidase Family II.
OKUYAMA MASAYUKI; MORI HARUHIDE; KIMURA ATSUO; CHIBA SEIYA
J Appl Glycosci, 49, 2, 211, 219, The Japanese Society of Applied Glycoscience, 01 Apr. 2002
Japanese, cDNA encoding Schizosaccharomyces pombe a-glucosidase was cloned, and expressed in Saccharomyces cerevisiae. The deduced amino acid sequence categorized under the α-glucosidase family II showed a high homology to those of a-glucosidase from molds, plants and mammals. By site direct mutagenesis, Asp481, G1u484, and Asp647 residues were confirmed to be essential in the catalytic reaction. The carboxyl group (-COON) of the Asp647 residue was for the first time pointed out to be the candidate of proton donor in the a-glucosidase of family II. The carboxylate group (-COO-) of the Asp481 residue was assumed to be the secondary carboxylate group, which stabilize the oxocarbenium ion through electrostatic interaction, and the Asp481 was considered to be modified by the chemical modification with conduritol B epoxide. The role of the G1u484 residue, which was the third residue, was presumed to be to fix the reaction intermediate of substrates.

Catalytic Amino Acid Residue Providing Proton Donor in α-Glucosidase Family II
OKUYAMA Masayuki; MORI Haruhide; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 49, 2, 211, 219, The Japanese Society of Applied Glycoscience, 01 Apr. 2002
Japanese, cDNA encoding Schizosaccharomyces pombe a-glucosidase was cloned, and expressed in Saccharomyces cerevisiae. The deduced amino acid sequence categorized under the α-glucosidase family II showed a high homology to those of a-glucosidase from molds, plants and mammals. By site direct mutagenesis, Asp481, G1u484, and Asp647 residues were confirmed to be essential in the catalytic reaction. The carboxyl group (-COON) of the Asp647 residue was for the first time pointed out to be the candidate of proton donor in the a-glucosidase of family II. The carboxylate group (-COO-) of the Asp481 residue was assumed to be the secondary carboxylate group, which stabilize the oxocarbenium ion through electrostatic interaction, and the Asp481 was considered to be modified by the chemical modification with conduritol B epoxide. The role of the G1u484 residue, which was the third residue, was presumed to be to fix the reaction intermediate of substrates.

Study on Three α-Glucosidase Isozymes from Honeybee, Apis mellifera L.
NISHIMOTO Mamoru; MORI Haruhide; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 49, 2, 191, 197, The Japanese Society of Applied Glycoscience, 01 Apr. 2002
Japanese, The genes of three a-glucosidases (HBG I, HBG II and HBG III) were isolated from the cDNA library of honeybee, Apis mellifera L. The nucleotide sequences of HBG I, II and III were consisted of 1974, 1910 and 1916 base pair and encoding 588, 580 and 567 amino acid residues, respectively. The putative primary structures showed high homology ranging from N- to C-terminals, and three enzymes belonged to a-glucosidase family I, in which four conservative regions of aamylase family were observed in their sequences. To obtain the recombinant enzymes, we tried to express the cDNAs in Pichia pastoris of heterologous host cells. Although recombinant HBG I was not produced, the recombinant HBG II and III of 2.4 and 1.2 U/mg were respectively expressed and secreted into culture supernatant. The active recombinant enzymes purified had the same properties as those of native ones except sugar content. To investigate the catalytic residues in HBGs, four mutated enzymes (D206N, E259Q, E269Q and D33 1N) of HBG III were constructed, and their specific activities were found to be 0.0004, 4.9, 0.004 and 0.0002 U/mg, respectively. E259Q remained half activity of wild type and those of the others disappeared, implying that three catalyticresidues of HBGs were D212, E281 and D343 of HBG I, D202, E271 and D333 of HBG II, D206, E269 and D331 of HBG III. The homology modeling showed that three enzymes had Ndomain ((β/α)₈ barrel), subdomain, and C-domain(β-sheet structure mainly) like oligo-l, 6-glucosidase from Bacillus cereus.

alpha-glucosidase mutant catalyzes "alpha-glycosynthase"-type reaction
M Okuyama; H Mori; K Watanabe; A Kimura; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66, 4, 928, 933, Apr. 2002
English, Scientific journal

Nishimoto M, Mori H, Kimura A & Chiba S, "Study on Three a-Glucosidase Isozymes from Honeybee, Apis mellifera L." J. Appl. Glycosci., 49(2) 191-197, 2002.
2002

Okuyama M, Mori H, Kimura A & Chiba S, "Catalytic Amino Acid Residue Providing Proton Donar in alpha-Glucosidase Family II" J.Appl.Glycosci., 49(2) 211-219, 2002
2002

Svensson B, Sauer J, Mori H, Jensen MT, Bak-Jensen KS, Kramhoft B, Juge N, Nohr J, Greffe L, Framdsem TP, Palcic MM, Williamson G, and Driguez H, "(Gluco)amylases, what have we learned so far ?" Carbohydrate Bioengineering, 67-75, 2002.
2002

Kinetic Studies on Substrate Specificity and Active Site of .BETA.-D-Glucosidase F1 from Streptomyces sp.
Fukuda Kenji; Shirakawa Kou; Mori Haruhide; Okuyama Masayuki; Kimura Atsuo; Ozaki Hachiro; Yoneyama Michio; Chiba Seiya
Journal of Applied Glycoscience, 49, 3, 265, 272, The Japanese Society of Applied Glycoscience, 2002
English, The substrate specificity and the active site of β-D-glucosidase F<SUP>-1</SUP> (Mr, 50, 000; optimum pH, 5.5) purified from a Streptomyces sp. were kinetically investigated. The β-D-glucosidase showed a broad substrate specificity for synthetic glycosides and disaccharides having β-glycosidic linkage, but the former was more favorable substrate than the latter. The enzyme was characterized by the ability to hydrolyze rapidly not only ρ-nitrophenyl β-glucoside (K<SUB>m</SUB>, 0.72 mM; k<SUB>0</SUB>, 63 s<SUP>-1</SUP>) but also ρ-nitrophenyl β-fucoside (K<SUB>m</SUB>, 0.19 mM; k<SUB>0</SUB>,...

(Gluco)amylases, what have we learned so far?
B Svensson; J Sauer; H Mori; MT Jensen; KS Bak-Jensen; B Kramhoft; N Juge; J Nohr; L Greffe; TP Frandsen; MM Palcic; G Williamson; H Driguez
CARBOHYDRATE BIOENGINEERING: INTERDISCIPLINARY APPROACHES, 275, 67, 75, 2002, [Peer-reviewed]
English, International conference proceedings

Purification and identification of the essential ionizable groups of honeybee, Apis mellifera L., trehalase
JH Lee; M Tsuji; M Nakamura; M Nishimoto; M Okuyama; H Mori; A Kimura; H Matsui; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 65, 12, 2657, 2665, Dec. 2001
English, Scientific journal

Isolation and Sequence of a Putative α-Glucosidase Gene from Brevibacterium fuscum var. dextranlyticum Strain 0407
MIZUNO Takafumi; MORI Haruhide; NISHIMOTO Mamoru; ITO Hiroyuki; MATSUI Hirokazu; KIMURA Atsuo; HONMA Mamoru; CHIBA Seiya
Journal of applied glycoscience, 48, 3, 287, 291, The Japanese Society of Applied Glycoscience, 01 Jul. 2001
English, A putative α-glucosidase gene was isolated from the genomic library of Brevibacterium fuscumvar. dextranlyticum strain 0407. The gene, designated dexG, was located upstream of isomaltotriodextranase gene (dexT). The dexG contained an open reading frame of 1725 bp, and its deduced amino acid sequence (DexG) showed a high homology with the enzymes belonging to α-glucosidase Family I and I-like, especially oligo-l, 6-glucosidase from Bacillus sp. and dextran glucosidase from Streptococcus mutans. The DexG has four conserved regions shared with aamylases. In the cloned genomic fragment there were two other open reading frames, of which the deduced amino acid sequences showed a similarity with those of oligosaccharides membrane transporter proteins. The gene cluster consisting of the membrane transporter protein genes, dexG, and dexT, seems to participate in the degradation and utilization of dextran in this bacterium.

Purification and substrate specificity of honeybee, Apis mellifera L., alpha-glucosidase III
M Nishimoto; M Kubota; M Tsuji; H Mori; A Kimura; H Matsui; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 65, 7, 1610, 1616, Jul. 2001
English, Scientific journal

Isolation and Sequence of a Putative .ALPHA.-Glucosidase Gene from Brevibacterium fuscum var. dextranlyticum Strain 0407.
Mizuno Takafumi; Mori Haruhide; Nishimoto Mamoru; Ito Hiroyuki; Matsui Hirokazu; Kimura Atsuo; Honma Mamoru; Chiba Seiya
Journal of Applied Glycoscience, 48, 3, 287, 291, The Japanese Society of Applied Glycoscience, 2001
English, A putative α-glucosidase gene was isolated from the genomic library of Brevibacterium fuscumvar. dextranlyticum strain 0407. The gene, designated dexG, was located upstream of isomaltotriodextranase gene (dexT). The dexG contained an open reading frame of 1725 bp, and its deduced amino acid sequence (DexG) showed a high homology with the enzymes belonging to α-glucosidase Family I and I-like, especially oligo-l, 6-glucosidase from Bacillus sp. and dextran glucosidase from Streptococcus mutans. The DexG has four conserved regions shared with aamylases. In the cloned genomic fragment there we...

Molecular cloning of isomaltotrio-dextranase gene from Brevibacterium fuscum var. dextranlyticum strain 0407 and its expression in Escherichia coli
T Mizuno; H Mori; H Ito; H Matsui; A Kimura; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 63, 9, 1582, 1588, Sep. 1999
English, Scientific journal

Localization of α-Glucosidase in Yeast Cells
SAEKI Takeshi; OKUYAMA Masayuki; MORI Haruhide; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 45, 3, 281, 283, 日本応用糖質科学会, 31 Aug. 1998
English

Molecular Cloning of an α-Amylase cDNA from Germinating Cotyledons of Kidney Bean (Phaseolus vulgaris L. cv. Toramame)
MORI Haruhide; KOBAYASHI Tetsuya; TONOKAWA Takashi; TATEMATSU Ayumi; MATSUI Hirokazu; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 45, 3, 261, 267, 日本応用糖質科学会, 31 Aug. 1998
English

Amylases and Branching Enzyme of Developing Kidney Bean Seeds on Native Electrophoretic Gel
NOZAKI Kouichi; MATSUI Hirokazu; TONOKAWA Takashi; MORI Haruhide; ITO Hiroyuki; HONMA Mamoru; CHIBA Seiya
Journal of applied glycoscience, 45, 2, 117, 122, 日本応用糖質科学会, 30 Jun. 1998
English

A catalytic amino acid and primary structure of active site in Aspergillus niger alpha-glucosidase
A Kimura; M Takata; Y Fukushi; H Mori; H Matsui; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 7, 1091, 1098, Jul. 1997
English, Scientific journal

Substrate Specificity and Primary Structure of Sugar Beet α-Glucosidase
MATSUI Hirokazu; IWANAMI Shunsuke; ITO Hiroyuki; KIMURA Atsuo; MORI Haruhide; HONMA Mamoru; CHIBA Seiya
Journal of applied glycoscience, 44, 2, 245, 252, 日本応用糖質科学会, 30 Jun. 1997
Japanese

Primary Structure and Sugar Chains of Crystalline α-Glucosidase from Aspergillus niger
KIMURA Atsuo; TAKAYANAGI Tsutomu; MORI Haruhide; MATSUI Hirokazu; UOZUMI Takeshi; CHIBA Seiya
Journal of applied glycoscience, 44, 2, 233, 243, 日本応用糖質科学会, 30 Jun. 1997
Japanese

Substrate Specificity and Primary Structure of Sugar Beet .ALPHA.-Glucosidase.
MATSUI HIROKAZU; IWANAMI SHUNSUKE; ITO HIROYUKI; KIMURA ATSUO; MORI HARUHIDE; HONMA MAMORU; CHIBA SEIYA
応用糖質科学, 44, 2, 245-252, Jun. 1997
Japanese

Primary Structure and Sugar Chains of Crystalline .ALPHA.-Glucosidase from Aspergillus niger.
KIMURA ATSUO; TAKAYANAGI TSUTOMU; MORI HARUHIDE; UOZUMI TAKESHI; CHIBA SEIYA; MATSUI HIROKAZU
応用糖質科学, 44, 2, 233-243, Jun. 1997
Japanese

Cloning and sequencing of a cDNA encoding alpha-glucosidase from sugar beet
H Matsui; S Iwanami; H Ito; H Mori; M Honma; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 5, 875, 880, May 1997
English, Scientific journal

Identification of essential ionizable groups in active site of Aspergillus niger alpha-glucosidase
A Kimura; A Somoto; H Mori; O Sakai; H Matsui; S Chiba
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 3, 475, 479, Mar. 1997
English, Scientific journal

Molecular cloning and nucleotide sequences of cDNA and gene encoding endo-inulinase from Penicillium purpurogenum
S Onodera; T Murakami; H Ito; H Mori; H Matsui; M Honma; S Chiba; N Shiomi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 60, 11, 1780, 1785, Nov. 1996
English, Scientific journal

Reinvestigation of Isomaltotrio-Dextranase from Brevibacterium fuscum var. dextranlyticum
MIZUNO Takafumi; MATSUI Hirokazu; ITO Hiroyuki; MORI Haruhide; KIMURA Atsuo; HONMA Mamoru; CHIBA Seiya
Journal of applied glycoscience, 43, 3, 347, 353, 日本応用糖質科学会, 31 Aug. 1996
English

Substrate Specificity and Subsite Affinities of α-Amylase from Germinating Cotyledons of Phaseolus vulgaris L. cv Toramame
MORI Haruhide; TATEMATSU Ayumi; SAITO Akiko; MATSUI Hirokazu; KIMURA Atsuo; CHIBA Seiya
Journal of applied glycoscience, 42, 4, 387, 394, 日本応用糖質科学会, 01 Dec. 1995
English

CHEMICAL MODIFICATION AND AMINO-ACID-SEQUENCE OF ACTIVE-SITE IN SUGAR-BEET ALPHA-GLUCOSIDASE
S IWANAMI; H MATSUI; A KIMURA; H ITO; H MORI; M HONMA; S CHIBA
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 3, 459, 463, Mar. 1995
English, Scientific journal

A sialic acid-binding lectin from the mashroom Hericium erinaceum.
Kawagishi H; Mori H; Uno A; Kimura A; Chiba S
FEBS Lett, 340, 1, 56, 58, 1994, [Peer-reviewed]
English, Scientific journal

STARCH-HYDROLYZING ENZYMES IN GERMINATING KIDNEY BEAN
H MORI; A TATEMATSU; H MATSUI; T TAKAYANAGI; M HONMA; S CHIBA
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 56, 9, 1499, 1500, Sep. 1992
English

NUCLEOTIDE AND DERIVED AMINO-ACID-SEQUENCE OF A CATALASE CDNA ISOLATED FROM RICE IMMATURE SEEDS
H MORI; K HIGO; H HIGO; Y MINOBE; H MATSUI; S CHIBA
PLANT MOLECULAR BIOLOGY, 18, 5, 973, 976, Mar. 1992
English

微生物による糖鎖の合成と分解 β1-3/1-6-グルカンに作用する微生物由来β1-3-グルカン分解酵素
太田智也; 佐分利亘; 森春英, 応用糖質科学, 15, 3, 2025

Bifidobacterium longum由来GH2 β-ガラクトシダーゼアイソザイム2種におけるラクトース分解過程でのGalβ1-6Glc蓄積量の差異とその機構
北見さわ子; 佐分利亘; 太田智也; 尾高伶; 堀口博文; 森春英, 日本応用糖質科学会大会・応用糖質科学シンポジウム講演要旨集, 74th-14th, 2025

ATP再生反応を用いたショ糖からのManβ1-4Glcの合成
山川佳乃; 泉しずく; 太田智也; 佐分利亘; 森春英, 日本応用糖質科学会大会・応用糖質科学シンポジウム講演要旨集, 74th-14th, 2025

GH55エンドβ-1,3-グルカナーゼにおけるサブサイト-1より基質の非還元末端側に延びるクレフトのラミナリン特異性への寄与
太田智也; 佐分利亘; 今場司朗; 今井亮三; 森春英, 日本応用糖質科学会大会・応用糖質科学シンポジウム講演要旨集, 74th-14th, 2025

β1-3グルカン分解酵素群の多様な切断様式とその分子基盤 ラミナリンを端から切る酵素と内から切る酵素の共通点と相違点
太田 智也; 佐分利 亘; 森 春英, 化学と生物, 62, 8, 362, 364, Aug. 2024, [Last author]
Introduction scientific journal

Bifidobacterium longum105A株由来GH2 β-ガラクトシダーゼの糖転移産物の解析
北見さわ子; 奥野剛基; 太田智也; 佐分利亘; 森春英, 日本農芸化学会北海道支部学術講演会講演要旨集(CD-ROM), 2024, 2024

GH130β-マンノシルグルコースホスホリラーゼの反応における基質非還元末端マンノシル基2位水酸基の寄与
服部駿太; 太田智也; 佐分利亘; 森春英, 日本農芸化学会北海道支部学術講演会講演要旨集(CD-ROM), 2024, 2024

ミツバチα-アミラーゼの構造と機能
佐分利亘; 高橋有志; 武井梓穂; 尾瀬農之; 森春英, 日本応用糖質科学会大会・応用糖質科学シンポジウム講演要旨集, 73rd-13th, 2024

新規酵素ガラクトース2-エピメラーゼの諸性質ならびにX線結晶構造の解析
内山昌典; 佐分利亘; 武井梓穂; 尾瀬農之; 森春英, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2023, 2023

GH55β-1,3-グルカナーゼのエンド様式の作用と構造
太田智也; 佐分利亘; 山下恵太郎; 田上貴祥; 于健; 今場司朗; ジュウェルリンダ; シャントム; 今井亮三; 姚閔; 森春英, 応用糖質科学, 13, 1, 2023, [Invited]
Introduction scientific journal

Chemical synthesis of oligosaccharide with partial structure of β1-3/1-6 glucan.
太田智也; 佐分利亘; 今場司朗; 森春英, 日本農芸化学会大会講演要旨集(Web), 2023, 2023

GH3β-グルコシダーゼMnBG3Aの反応速度と基質阻害に対する単糖の影響
太田智也; 佐分利亘; JEWELL Linda; HSIANG Tom; 今井亮三; 森春英, 日本栄養・食糧学会北海道支部大会講演要旨集, 53rd (CD-ROM), 2023

雪腐病菌由来GH3β-グルコシダーゼMnBG3AのpNP-Glc加水分解速度への各種配糖体および単糖の影響
太田智也; 佐分利亘; JEWELL Linda; HSIANG Tom; 今井亮三; 森春英, 応用糖質科学, 13, 3, 2023

Laminarin degradation mechanism of GH55 laminarinase from Microdochium nivale
太田智也; 佐分利亘; 今場司朗; JEWELL Linda; HSIANG Tom; 今井亮三; 森春英, 日本農芸化学会大会講演要旨集(Web), 2022, 2022

GH55β-1,3-グルカナーゼのエンド様式の作用と構造
太田智也; 佐分利亘; 山下恵太郎; 田上貴祥; 于健; 今場司朗; JEWELL Linda; HSIANG Tom; 今井亮三; 姚閔; 森春英, 応用糖質科学, 12, 3, 2022

紅色雪腐病菌Microdochium nivale由来GH3β-グルコシダーゼの酵素化学的諸性質とオリゴ糖の合成
太田智也; 佐分利亘; JEWELL Linda; HSIANG Tom; 今井亮三; 森春英, 応用糖質科学, 10, 4, 2020

Functional analysis of the extracellular β-glucosidase from phytopathogenic fungus Microdochium nivale
太田智也; 佐分利亘; JEWELL Linda; HSIANG Tom; 今井亮三; 森春英, 日本農芸化学会大会講演要旨集(Web), 2020, 2020

植物病原性真菌Microdochium nivale由来菌体外ラミナリナーゼの機能解析
太田智也; 佐分利亘; 今井亮三; 森春英, 応用糖質科学, 9, 3, 2019

ビフィズス菌のトランスポゾン変異株を用いたα-グルコシダーゼ遺伝子の機能解明
阪中幹祥; 阪中幹祥; 阪中幹祥; 中川路伸吾; 中島森; 阿部光紗; 佐分利亘; 森春英; 横田篤; 吹谷智, 日本農芸化学会大会講演要旨集(Web), 2019, 2019

セロビオース2‐エピメラーゼによるマンノビオースのエピメリ化機構
佐分利亘; 加藤公児; 于健; 姚閔; 森春英, 応用糖質科学, 8, 3, (40), 20 Aug. 2018
Japanese

ゲノム編集を用いたサリチル酸高蓄積イネ変異体の作出
手塚大介; 手塚大介; 佐分利亘; 森春英; 松浦英幸; 今井亮三, 植物の生長調節, 53, Supplement, 2018

転写因子に着目したトレハロース誘導抵抗性のシグナル伝達機構の解析
手塚大介; 手塚大介; 川又彩; 加藤英樹; 佐分利亘; 森春英; 今井亮三; 今井亮三, 日本植物細胞分子生物学会大会・シンポジウム講演要旨集, 36th, 2018

Ten years of CAZypedia: a living encyclopedia of carbohydrate-active enzymes
Wade Abbott; Orly Alber; Ed Bayer; Jean-Guy Berrin; Alisdair Boraston; Harry Brumer; Ryszard Brzezinski; Anthony Clarke; Beatrice Cobucci-Ponzano; Darrell Cockburn; Pedro Coutinho; Mirjam Czjzek; Bareket Dassa; Gideon John Davies; Vincent Eijsink; Jens Eklof; Alfons Felice; Elizabeth Ficko-Blean; Geoff Pincher; Thierry Fontaine; Zui Fujimoto; Kiyotaka Fujita; Shinya Fushinobu; Harry Gilbert; Tracey Gloster; Ethan Goddard-Borger; Ian Greig; Jan-Hendrik Hehemann; Glyn Hemsworth; Bernard Henrissat; Masafumi Hidaka; Ramon Hurtado-Guerrero; Kiyohiko Igarashi; Takuya Ishida; Stefan Janecek; Seino Jongkees; Nathalie Juge; Satoshi Kaneko; Takane Katayama; Motomitsu Kitaoka; Naotake Konno; Daniel Kracher; Anna Kulminskaya; Alicia Lammerts van Bueren; Sine Larsen; Junho Lee; Markus Linder; Leila LoLeggio; Roland Ludwig; Ana Luis; Mirko Maksimainen; Brian Mark; Richard McLean; Gurvan Michel; Gurvan Michel; Cedric Montanier; Marco Moracci; Haruhide Mori; Hiroyuki Nakai; Wim Nerinckx; Takayuki Ohnuma; Richard Pickersgill; Kathleen Piens; Tirso Pons; Etienne Rebuffet; Peter Reilly; Magali Remaud-Simeon; Brian Rempel; Kyle Robinson; David Rose; Juha Rouvinen; Wataru Saburi; Yuichi Sakamoto; Mats Sandgren; Fathima Shaikh; Yuval Shoham; Franz St John; Jerry Stahlberg; Michael Suits; Gerlind Sulzenbacher; Tomomi Sumida; Ryuichiro Suzuki; Birte Svensson; Toki Taira; Ed Taylor; Takashi Tonozuka; Breeanna Urbanowicz; Gustav Vaaje-Kolstad; Wim Van den Ende; Annabelle Varrot; Maxime Versluys; Florence Vincent; David Vocadlo; Warren Wakarchuk; Tom Wennekes; Rohan Williams; Spencer Williams; David Wilson; Stephen Withers; Katsuro Yaoi; Vivian Yip; Ran Zhang, GLYCOBIOLOGY, 28, 1, 3, 8, Jan. 2018
English, Others

アルドース‐ケトース間の異性化反応を利用したセロビオース2‐エピメラーゼの反応機構の解析
武藤洋彦; 佐分利亘; 藤原孝彰; 加藤公児; YAO Min; 森春英, 日本農芸化学会大会講演要旨集(Web), 2017, ROMBUNNO.3J33p07 (WEB ONLY), 05 Mar. 2017
Japanese

イネのトレハロース誘導抵抗性に関与する新たなERF転写因子の機能解析
手塚大介; 手塚大介; 川又彩; 加藤英樹; 佐分利亘; 森春英; 今井亮三, 植物の生長調節, 51, Supplement, 59, 07 Oct. 2016
Japanese

トレハロース6‐リン酸ホスホリラーゼを用いたトレハロース6‐リン酸および新規糖リン酸の合成
田口陽大; 佐分利亘; 今井亮三; 森春英, 応用糖質科学, 6, 3, 46, 20 Aug. 2016
Japanese

β‐マンナン分解に寄与するセロビオース2‐エピメラーゼとβ‐マンノシドホスホリラーゼの構造と機能
佐分利亘; 加藤公児; 姚閔; 松井博和; 森春英, 応用糖質科学, 6, 3, 65, 65, 20 Aug. 2016
(一社)日本応用糖質科学会, Japanese

マルトースを出発物質とするトレハロース6‐リン酸の酵素合成法の確立
田口陽大; 佐分利亘; 今井亮三; 今井亮三; 森春英, 日本農芸化学会大会講演要旨集(Web), 2016, 4D028 (WEB ONLY), 05 Mar. 2016
Japanese

乳中ラクトースのエピラクトースへの直接変換を指向した各種セロビオース2‐エピメラーゼの機能評価
武藤洋彦; 佐分利亘; 森春英, 日本農芸化学会大会講演要旨集(Web), 2016, 4D008 (WEB ONLY), 05 Mar. 2016
Japanese

Gluconobacter oxydans由来の菌体外酵素dextran dextrinaseは自らの生成物デキストランによって細胞表層から遊離する.
貞廣樹里; 森春英; 佐分利亘; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集(Web), 2016, 4D013 (WEB ONLY), 05 Mar. 2016
Japanese

嫌気性細菌によるエピラクトース代謝の多様性
坂井未悠; 佐分利亘; 森春英, 日本農芸化学会大会講演要旨集(Web), 2016, 2E063 (WEB ONLY), 05 Mar. 2016
Japanese

Ruminococcus albus由来4‐O‐β‐D‐マンノシル‐D‐グルコースホスホリラーゼのサブサイト+1に保存されるArg92の基質結合への関与
塩田咲耶子; 尾高伶; 佐分利亘; YE Yuxin; 薦田圭介; 加藤公児; 西本完; 北岡本光; YAO Min; 森春英, 日本農芸化学会大会講演要旨集(Web), 2016, 4D009 (WEB ONLY), 05 Mar. 2016
Japanese

P055 Molecular basis of Trehalose-induced Systemic Resistance
Tezuka Daisuke; Wakuta Shinji; Kato Hideki; Matsuura Hideyuki; Saburi Wataru; Mori Haruhide; Matsui Hirokazu; Imai Ryozo, 植物化学調節学会研究発表記録集, 50, 0, 73, 73, 01 Oct. 2015
The Japanese Society for Chemical Regulation of Plants, Japanese

Trehalose‐induced Systemic Resistance(TSR)の分子基盤
手塚大介; 手塚大介; 和久田真司; 加藤英樹; 松浦英幸; 佐分利亘; 森春英; 松井博和; 今井亮三; 今井亮三, 植物の生長調節, 50, Supplement, 73, 01 Oct. 2015
Japanese

S3-2 長鎖阻害剤の利用による植物α-グルコシダーゼの機能構造相関の解明(応用糖質科学シンポジウム,一般社団法人日本応用糖質科学会平成27年度大会(第64回))
田上 貴祥; 山下 恵太郎; 奥山 正幸; 森 春英; 姚 閔; 木村 淳夫, 応用糖質科学 : 日本応用糖質科学会誌, 5, 3, B59, 20 Aug. 2015
日本応用糖質科学会, Japanese

Ba-3 Isomaltooligosaccharide 6-α-glucosyltransferaseの構造と触媒部位の機能性アミノ酸残基の同定(澱粉関連酵素,一般講演,一般社団法人日本応用糖質科学会平成27年度大会(第64回))
川田 恭平; 藤本 瑞; 鐘ケ江 倫世; 西村 崇志; 奥山 正幸; 森 春英; 木村 淳夫, 応用糖質科学 : 日本応用糖質科学会誌, 5, 3, B41, 20 Aug. 2015
日本応用糖質科学会, Japanese

Ba-9 Ruminococcus albus由来4-O-β-D-mannosyl-D-glucose phosphorylase RaMP1の無機リン酸に対するホモトロピック相互作用(ホスホリラーゼ関連,一般講演,一般社団法人日本応用糖質科学会平成27年度大会(第64回))
尾高 伶; 佐分 利亘; Ye Yuxin; 薦田 圭介; 加藤 公児; 西本 完; 北岡 本光; 姚 閔; 森 春英, 応用糖質科学 : 日本応用糖質科学会誌, 5, 3, B42, 20 Aug. 2015
日本応用糖質科学会, Japanese

Bacillus sp.AHU2001由来GH31α‐グルコシダーゼの生化学的諸性質と基質認識に重要な構造因子の解析
佐分利亘; 奥山正幸; 熊谷祐也; 木村淳夫; 森春英, 日本農芸化学会大会講演要旨集(Web), 2015, 3E32P02 (WEB ONLY), 05 Mar. 2015
Japanese

Rhodothermus marinus JCM9785由来セロビオース2‐エピメラーゼの基質結合部位周辺アミノ酸残基の機能解析
武藤洋彦; 佐分利亘; 藤原孝彰; YAO Min; 森春英, 日本農芸化学会大会講演要旨集(Web), 2015, 2E32A08 (WEB ONLY), 05 Mar. 2015
Japanese

グライコシンターゼ反応を利用したシロイヌナズナ由来β‐グルコシダーゼBglu15のアグリコン特異性の解析
菅原好美; 佐分利亘; 谷口沙希; 今井亮三; 森春英, 日本農芸化学会大会講演要旨集(Web), 2015, 2E32A11 (WEB ONLY), 05 Mar. 2015
Japanese

テンサイα-グルコシダーゼの高重合度基質特異性はらせん構造の長鎖基質に適したサブサイト構造に起因する
田上貴祥; 山下恵太郎; 奥山正幸; 森春英; YAO Min; YAO Min; 木村淳夫, 日本農芸化学会大会講演要旨集(Web), 2015, 2015

19. Reduction of GA levels in response to chilling enhances chilling tolerance of rice
Takahashi Naoki; Kawamata Aya; Tezuka Daisuke; Saburi Wataru; Matsuura Hideyuki; Mori Haruhide; Imai Ryozo, 植物化学調節学会研究発表記録集, 49, 0, 37, 37, 01 Oct. 2014
植物化学調節学会, Japanese

43. Glucosylation of tuberonic acid using salicylic acid glucoside in wound rice
Takematsu Tomonori; Seto Yoshiya; Miyazawa Yoshiroh; Wakuta Shinji; Saburi Wataru; Mori Haruhide; Takahashi Kosaku; Matsuura Hideyuki, 植物化学調節学会研究発表記録集, 49, 0, 61, 61, 01 Oct. 2014
植物化学調節学会, Japanese

52. Jasmonic acid is involved in the trehalose-induced systemic disease resistance in rice
Tezuka Daisuke; Sakai Shiho; Wakuta Shinji; Kato Hideki; Matsuura Hideyuki; Saburi Wataru; Mori Haruhide; Matsui Hirokazu; Imai Ryozo, 植物化学調節学会研究発表記録集, 49, 0, 70, 70, 01 Oct. 2014
植物化学調節学会, Japanese

62. A possible function of a salicylic acid β-glucosidase in rice wound response
Takeda Ryosuke; Saburi Wataru; Himeno Nami; Wakuta Shinji; Matsuura Hideyuki; Imai Ryozo; Matsui Hirokazu; Mori Haruhide, 植物化学調節学会研究発表記録集, 49, 0, 80, 80, 01 Oct. 2014
植物化学調節学会, Japanese

イネの傷害応答におけるサリチル酸グルコシド加水分解酵素の機能
武田遼介; 佐分利亘; 姫野奈美; 和久田真司; 松浦英幸; 今井亮三; 松井博和; 森春英, 植物の生長調節, 49, Supplement, 80, 01 Oct. 2014
Japanese

イネのトレハロースにより誘導されるシステミックな病害抵抗性にはジャスモン酸が関与する
手塚大介; 坂井志帆; 和久田真司; 加藤英樹; 松浦英幸; 佐分利亘; 森春英; 松井博和; 今井亮三, 植物の生長調節, 49, Supplement, 70, 01 Oct. 2014
Japanese

傷害時のイネにおけるサリチル酸グリコシドを糖ドナーとするツベロン酸への糖転移反応
竹松知紀; 瀬戸義哉; 宮澤吉郎; 和久田真司; 佐分利亘; 森春英; 高橋公咲; 松浦英幸, 植物の生長調節, 49, Supplement, 61, 01 Oct. 2014
Japanese

低温に応答したGA量の抑制によりイネの耐性を高めることができる
高橋直希; 川又彩; 手塚大介; 佐分利亘; 松浦英幸; 森春英; 今井亮三, 植物の生長調節, 49, Supplement, 37, 01 Oct. 2014
Japanese

Halomonas sp. H11株由来α‐グルコシダーゼの糖転移機構の解析
城戸悠輔; 佐分利亘; 小島晃代; SHEN Xing; 薦田圭介; 姚閔; 松井博和; 森春英, 日本農芸化学会北海道支部講演会講演要旨, 2014, 32, 22 Sep. 2014
Japanese

Bp-6 Amphiphilic function of linear-isomaltomegalosaccharides (L-IMS) on ethyl red (ER) solubility
Lang Weeranuch; Kumagai Yuya; Sadahiro Juri; Okuyama Masayuki; Mori Haruhide; Sakairi Nobuo; Kimura Atsuo, Bulletin of applied glycoscience, 4, 3, B42, 20 Aug. 2014
The Japanese Society of Applied Glycoscience, English

イネ由来スクロースシンターゼ3のヌクレオチド二リン酸特異性に関わるアミノ酸残基の決定
岩藤伸治; 佐分利亘; 松井博和; 今井亮三; 森春英, 応用糖質科学, 4, 3, (40), 20 Aug. 2014
日本応用糖質科学会, Japanese

難消化性二糖エピラクトースによる抗メタボリックシンドローム作用の解析
村上祐紀; 佐分利亘; 森春英; 松井博和; 田辺創一; 鈴木卓弥, 日本栄養・食糧学会大会講演要旨集, 68th, 257, 30 Apr. 2014
Japanese

Halomonas sp.H11株由来α‐グルコシダーゼD274G変異酵素の一価カチオンによる活性化機構の解析
城戸悠輔; 佐分利亘; 小島晃代; 松井博和; 森春英, 日本農芸化学会大会講演要旨集(Web), 2014, 2D02A05 (WEB ONLY), 05 Mar. 2014
Japanese

アクセプタ結合部位を改変したRuminococcus albus由来マンノシドホスホリラーゼの基質特異性と反応特性の解析
尾高伶; 佐分利亘; 福士江里; 西本完; 北岡本光; 松井博和; 森春英, 日本農芸化学会大会講演要旨集(Web), 2014, 3D02P09 (WEB ONLY), 05 Mar. 2014
Japanese

Ba-3 Dextran dextrinaseのPhe602の生成物特異性に関する影響(その他の糖質関連酵素,一般講演,日本応用糖質科学会平成26年度大会(第63回))
坂谷 敦; 熊谷 祐也; 貞廣 樹里; Weeranuch Lang; 奥山 正幸; 森 春英; 木村 淳夫, 応用糖質科学：日本応用糖質科学会誌, 4, 3, B39, 2014
一般社団法人 日本応用糖質科学会, Japanese

Acidophilic β-Galactosidase from Aspergillus niger AHU7120 with Lactose Hydrolytic Activity Under Simulated Gastric Conditions
Saburi Wataru; M. Ueno Hiroshi; Matsui Hirokazu; Mori Haruhide, Journal of Applied Glycoscience, 61, 2, 53, 57, 2014
Acidophilic β-galactosidase is a useful enzyme as digestive supplement used to alleviate symptoms of lactose intolerance. Aspergilli are the source of several acidophilic β-galactosidases that retain enzymatic activity under gastric conditions. In this study, we investigated the extracellular acidophilic β-galactosidase activity of six Aspergillus niger strains, AHU7104, AHU7120, AHU7217, AHU7294, AHU7295 and AHU7296; A. niger AHU7120 was selected as an enzyme source. β-Galactosidase from A. niger AHU7120 (AnBGal) was purified from culture supernatant. Its N-terminal sequence was identical to that of An01g12150, which belongs to the glycoside hydrolase family 35, from A. niger CBS 513.88. The DNA sequence of AnBGal was identical to An01g12150. Recombinant AnBGal (rAnBGal) harboring yeast α-factor signal sequence was expressed in Pichia pastoris, and 21.9 mg of purified rAnBGal with 129 U/mg of enzyme activity was isolated from 200 mL of culture supernatant. Native and recombinant AnBGal enzymes showed similar pH optima, pH stability, and kinetics for p-nitrophenyl β-D-galactopyranoside and lactose; rAnBGal showed slightly lower thermal stability than the native enzyme. Lactose in milk was rapidly degraded by rAnBGal at higher pH values (range, 2.0‒3.5), consistent with the pH optimum of AnBGal. We estimated that 3.5 μM AnBGal may degrade ≥ 66% of lactose before gastric half-emptying of ingested milk. These data indicate that AnBGal may help alleviate symptoms of lactose intolerance., The Japanese Society of Applied Glycoscience, English

Colorimetric Quantification of β-(1→4)-Mannobiose and 4-O-β-D-Mannosyl-D-glucose
Jaito Nongluck; Saburi Wataru; Muto Hirohiko; Matsui Hirokazu; Mori Haruhide, Journal of Applied Glycoscience, 61, 4, 117, 119, 2014
Spectrophotometric quantification method of carbohydrates is useful for processing multiple samples. In this study, we established colorimetric quantification for 4-O-β-D-mannosyl-D-glucose (Man-Glc) and β-(1→4)-mannobiose (Man2). For quantification of Man-Glc, phosphorolysis of Man-Glc catalyzed by 4-O-β-D-mannosyl-D-glucose phosphorylase (MGP) was coupled with quantification of D-glucose by the glucose oxidase-peroxidase method. In addition to MGP, cellobiose 2-epimerase (CE) was added for quantification of Man2. In both quantifications, a good linear relationship was obtained between A505 and the sample concentration (0-0.5 mM). The A505 values obtained at various concentrations of Man2 and Man-Glc were almost identical to those with equivalent D-glucose concentrations. Kinetic parameters of Ruminococcus albus and Rhodothermus marinus CEs for the epimerization of Man2 were determined using the quantification method for Man-Glc. Both enzymes showed 5-15-fold higher kcat/Km values than those for cellobiose and lactose, which supports the prediction that these enzymes utilize Man2 as a substrate in the β-mannan metabolic pathway., The Japanese Society of Applied Glycoscience, English

シロイヌナズナβ‐グルコシダーゼ様タンパク質の生化学的機能解析
谷口沙希; 佐分利亘; 松浦英幸; 今井亮三; 松井博和; 森春英, 日本農芸化学会北海道支部講演会講演要旨, 2013, 36, 27 Nov. 2013
Japanese

Gluconobacter oxydansが産生する2つのデキストラン合成酵素の同一性に関する解析
貞廣樹里; 森春英; 佐分利亘; 奥山正幸; 木村淳夫, 日本農芸化学会北海道支部講演会講演要旨, 2013, 17, 27 Nov. 2013
Japanese

イネ胚乳ADP‐グルコースピロホスホリラーゼの酵素化学的諸性質
石塚佐都子; 和久田真司; 佐分利亘; 今井亮三; 森春英, 日本農芸化学会北海道支部講演会講演要旨, 2013, 23, 27 Nov. 2013
Japanese

シロイヌナズナβ‐グルコシダーゼ様タンパク質の生化学的機能解析
谷口沙希; 佐分利亘; 松浦英幸; 今井亮三; 松井博和; 森春英, 日本農芸化学会北海道支部講演会講演要旨, 2013, 23, 27 Nov. 2013
Japanese

Cellvibrio vulgaris NCIMB8633のマンナン代謝オペロンの機能解析
田中佑果; 佐分利亘; 森春英, 日本農芸化学会北海道支部講演会講演要旨, 2013, 25, 27 Nov. 2013
Japanese

Ca-2 Schizosaccharomyces pombe由来グルコシダーゼIIα-サブユニットの大腸菌生産および機能解析(α-グルコシダーゼ・その他の糖質関連酵素1,一般講演,日本応用糖質科学会平成25年度大会(第62回))
宮本 大司; 奥山 正幸; 松尾 一郎; 森 春英; 木村 淳夫, 応用糖質科学 : 日本応用糖質科学会誌, 3, 3, B37, 20 Aug. 2013
日本応用糖質科学会, Japanese

Cp-15 糖鎖の長さを制御できる新規両親媒性プライマーの調製(多糖の構造・物性・合成,一般講演,日本応用糖質科学会平成25年度大会(第62回))
西堀 貴哉; 鈴木 志保; 森 春英; 木村 淳夫; 秋吉 一成; 北村 進一, 応用糖質科学 : 日本応用糖質科学会誌, 3, 3, B43, 20 Aug. 2013
日本応用糖質科学会, Japanese

Ca-3 Streptococcus mutans由来dextran glucosidaseのβ→α loop 6への変異導入による鎖長特異性の改変(α-グルコシダーゼ・その他の糖質関連酵素1,一般講演,日本応用糖質科学会平成25年度大会(第62回))
小林 桃子; 森 春英; 奥山 正幸; 木村 淳夫, 応用糖質科学 : 日本応用糖質科学会誌, 3, 3, B38, 20 Aug. 2013
日本応用糖質科学会, Japanese

Halomonas sp.H11株由来α‐グルコシダーゼの一価陽イオンによる活性化機構の解析
城戸悠輔; 佐分利亘; 小島晃代; 松井博和; 森春英, 応用糖質科学, 3, 3, 37, 20 Aug. 2013
日本応用糖質科学会, Japanese

Bacillus sp.AAH‐31株由来α‐アミラーゼの一次構造の解析とN末端ドメインの機能解析
佐分利亘; 森本奈保喜; 向井惇; KIM Dae Hoon; 竹花稔彦; 小池誠治; 松井博和; 森春英, 応用糖質科学, 3, 3, 38, 20 Aug. 2013
日本応用糖質科学会, Japanese

セロビオース2‐エピメラーゼの異性化反応機構の解明
藤原孝彰; 佐分利亘; 松井博和; 森春英; 田中勲, 姚閔, 応用糖質科学, 3, 3, 30, 20 Aug. 2013
Japanese

セロビオース2‐エピメラーゼを用いたエピラクトースの実用的酵素合成法の開発
佐分利亘; 小島晃代; 佐藤央基; 田口秀典; 森春英; 松井博和, 応用糖質科学, 3, 2, 137, 142, 20 May 2013
セロビオース2-エピメラーゼ(CE)は,セロビオースやラクトースなどβ-1,4結合からなるオリゴ糖の還元末端のグルコース残基をマンノース残基に可逆的に異性化する。CEをラクトースに作用させて得られるエピラクトース(4-O-β-D-ガラクトシル-D-マンノース)は優れた腸内細菌叢改善効果を有し,新たな食品素材として有望である。本研究では,CEを用いたエピラクトースの実用的合成法の確立を目的とした。Ruminococcus albus由来CEのアミノ酸配列を基に配列類似性検索を行い,大規模培養が可能な好気性細菌よりCEホモログを探索した。見出されたCEのうちRhodothermus marinus由来酵素(RmCE)は耐熱性に優れ,エピラクトースの工業的製造に適した特性を備えていた。固定化RmCEによる連続反応では,遊離酵素よりも少ない酵素量でエピラクトースを合成可能なことが示された。反応液を濃縮してラクトースを結晶化,除去した後,樹脂分画もしくは結晶化を行うことでCE反応液からエピラクトースを約90%の純度に精製した。, 日本応用糖質科学会, Japanese

Megalo types of alpha-1,6-glucosaccharide enhance absorption of quercetin glycosides in rats
Aki Shinoki; Weeranuch Lang; Haruhide Mori; Atsuo Kimura; Satoshi Ishizuka; Hiroshi Hara, FASEB JOURNAL, 27, Apr. 2013
English, Summary international conference

イネ由来ツベロン酸β‐グルコシドグルコシダーゼのアグリコン認識メカニズムの解析
姫野奈美; 佐分利亘; 武田遼介; 和久田真司; 松浦英幸; 鍋田憲助; 森春英; 今井亮三; 松井博和, 日本農芸化学会大会講演要旨集(Web), 2013, 3C16A13 (WEB ONLY), 05 Mar. 2013
Japanese

デキストラン合成酵素dextran dextrinaseの触媒残基およびその機能の決定
貞廣樹里; 森春英; 佐分利亘; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集(Web), 2013, 2C16A12 (WEB ONLY), 05 Mar. 2013
Japanese

イネ由来ツベロン酸β‐グルコシドグルコシダーゼアイソザイム1の機能解析
武田遼介; 姫野奈美; 佐分利亘; 和久田真司; 森春英; 松浦英幸; 鍋田憲助; 今井亮三; 松井博和, 日本農芸化学会大会講演要旨集(Web), 2013, 3C16A12 (WEB ONLY), 05 Mar. 2013
Japanese

Halomonas sp.H11株由来α‐グルコシダーゼ組換え酵素の大腸菌による生産と諸性質
城戸悠輔; 佐分利亘; 小島晃代; 森春英; 松井博和, 日本農芸化学会大会講演要旨集(Web), 2013, 3C16A01 (WEB ONLY), 05 Mar. 2013
Japanese

Da-10 Gluconobacter oxydans由来dextran dextrinaseによる直鎖イソマルトメガロ糖の酵素合成(糖質の合成・生産,一般講演,日本応用糖質科学会平成25年度大会(第62回))
熊谷 祐也; Lang Weeranuch; 貞廣 樹里; 奥山 正幸; 森 春英; 木村 淳夫, 応用糖質科学：日本応用糖質科学会誌, 3, 3, B46, 2013
一般社団法人 日本応用糖質科学会, Japanese

Bacillus sp.AAH‐31株由来耐熱性アルカリα‐アミラーゼの高機能化に関する研究
玉村尚也; 向井惇; 森本奈保喜; 竹花稔彦; 佐分利亘; 森春英; 小池誠治; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2012, 24, 01 Nov. 2012
Japanese

ツベロン酸グルコシド加水分解酵素の基質認識機構の解明
姫野奈美; 和久田真司; 武田遼介; 佐分利亘; 森春英; 松浦英幸; 鍋田憲助; 今井亮三; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2012, 24, 01 Nov. 2012
Japanese

組換えツベロン酸グルコシドグルコシダーゼの酵素化学的諸性質の解析
武田遼介; 姫野奈美; 佐分利亘; 和久田真司; 森春英; 松浦英幸; 鍋田憲助; 今井亮三; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2012, 23, 01 Nov. 2012
Japanese

Bp1-10 cellobiose phosphorylaseのepilactose phosphorylaseへの改変(ホスホリラーゼおよび関連酵素,一般講演,日本応用糖質科学会平成24年度大会(第61回))
羽村 健; 佐分 利亘; 森 春英; 松井 博和, 応用糖質科学 : 日本応用糖質科学会誌, 2, 3, B37, 20 Aug. 2012
日本応用糖質科学会, Japanese

S3-4 セロビオース2-エピメラーゼを用いたエピラクトースの実用的酵素合成法の開発(応用糖質科学シンポジウム(旧糖質関連酵素化学シンポジウム),日本応用糖質科学会平成24年度大会(第61回))
佐分 利亘; 小島 晃代; 佐藤 央基; 田口 秀典; 森 春英; 松井 博和, 応用糖質科学 : 日本応用糖質科学会誌, 2, 3, B58, 20 Aug. 2012
日本応用糖質科学会, Japanese

Ruminococcus albus由来セロビオース 2‐エピメラーゼ(RaCE)のX線結晶構造解析
藤原孝彰; 佐分利亘; 井上聡太; 森春英; 松井博和; 姚閔, 田中勲, 応用糖質科学, 2, 3, (53), 20 Aug. 2012
日本応用糖質科学会, Japanese

Acarviosyl‐maltooligsaccharideの酵素合成とテンサイα‐glucosidaseに対する基質アナログとしての利用
田上貴祥; 山下恵太郎; 田中良幸; 奥山正幸; 森春英; 姚閔; 木村淳夫, 応用糖質科学, 2, 3, (34), 20 Aug. 2012
日本応用糖質科学会, Japanese

cellobiose phosphorylaseのepilactose phosphorylaseへの改変
羽村健; 佐分利亘; 森春英; 松井博和, 応用糖質科学, 2, 3, (37), 20 Aug. 2012
Japanese

セロビオース2‐エピメラーゼを用いたエピラクトースの実用的酵素合成法の開発
佐分利亘; 小島晃代; 佐藤央基; 田口秀典; 森春英; 松井博和, 応用糖質科学, 2, 3, (58), 20 Aug. 2012
Japanese

Ruminococcus albus由来セロデキストリンホスホリラーゼの諸性質とリン酸結合部位の解析
澤野達也; 佐分利亘; 森春英; 松井博和, 応用糖質科学, 2, 3, (38), 20 Aug. 2012
日本応用糖質科学会, Japanese

Ruminococcus albus NE1株由来マンノシルグルコースホスホリラーゼ(RaMP1)の一般酸触媒残基の解析
尾高伶; 佐分利亘; 川原良介; 森春英; 松井博和, 応用糖質科学, 2, 3, (38), 20 Aug. 2012
日本応用糖質科学会, Japanese

固定化セロビオース2‐エピメラーゼによるエピラクトースの合成
佐藤央基; 佐分利亘; 小島晃代; 田口秀典; 森春英; 松井博和, 日本農芸化学会大会講演要旨集(Web), 2012, 4C10A04 (WEB ONLY), 05 Mar. 2012
Japanese

Ruminococcus albus NE1株由来cellobiose phosphorylaseのアクセプター特異性の改変
羽村健; 佐分利亘; 森本奈保喜; 森春英; 松井博和, 日本農芸化学会大会講演要旨集(Web), 2012, 2C10P21 (WEB ONLY), 05 Mar. 2012
Japanese

Streptococcus mutans由来デキストラナーゼの結晶構造に基づく触媒機構の解明
鈴木喜大; 金泳みん; 金泳みん; 藤本瑞; 門間充; 奥山正幸; 森春英; 舟根和美; 木村淳夫, PFシンポジウム要旨集, 29th, 28, 2012
Japanese

Biochemical Characterization of a Thermophilic Cellobiose 2-Epimerase from a Thermohalophilic Bacterium, Rhodothermus marinus JCM9785
Teruyo Ojima; Wataru Saburi; Hiroki Sato; Takeshi Yamamoto; Haruhide Mori; Hirokazu Matsui, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 75, 11, 2162, 2168, Nov. 2011
English

デキストラングルコシダーゼとイソマルトオリゴ糖6‐α‐グルコシル転移酵素:基質認識と糖転移反応のメカニズム
森春英, 飯島記念食品科学振興財団年報, 2009, 348, Aug. 2011
Japanese

Calcium Ion-Dependent Increase in Thermostability of Dextran Glucosidase from Streptococcus mutans
Momoko Kobayashi; Hironori Hondoh; Haruhide Mori; Wataru Saburi; Masayuki Okuyama; Atsuo Kimura, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 75, 8, 1557, 1563, Aug. 2011
English

Ruminococcus albus NE1におけるマンナン資化性の解析とβ‐1,4‐マンナナーゼの同定
阪本安希; 中島碧; 田口秀典; 佐分利亘; 森春英; 松井博和, 応用糖質科学, 1, 3, (50), 20 Jul. 2011
Japanese

Ruminococcus albus NE1株由来cellobiose phosphorylaseの諸性質の解析
羽村健; 阿部正太郎; 河内慎平; 森本奈保喜; 佐分利亘; 森春英; 松井博和, 応用糖質科学, 1, 3, (51), 20 Jul. 2011
Japanese

Isomaltooligosaccharide 6‐α‐glucosyltransferaseサブサイト+1変異酵素の機能解析
青山泰; 西村崇志; 鐘ケ江倫世; 本同宏成; 奥山正幸; 森春英; 木村淳夫, 応用糖質科学, 1, 3, (36), 20 Jul. 2011
Japanese

Pichia pastorisによる組換えツベロン酸グルコシドβ‐glucosidaseの諸性質の解明
姫野奈美; 和久田真司; 佐分利亘; 森春英; 松浦英幸; 鍋田憲助; 今井亮三; 松井博和, 応用糖質科学, 1, 3, (47), 20 Jul. 2011
Japanese

好熱性好気性細菌Rhodothermus marinus JCM 9785株より単離したcellobiose 2‐epimeraseの諸性質
小島晃代; 佐分利亘; 佐分利亘; 山本健; 佐藤央基; 森春英; 松井博和, 応用糖質科学, 1, 3, (51), 20 Jul. 2011
Japanese

Dextran glucosidase(DexB)ウォーターパス改変体の機能と構造
小林桃子; 山下恵太郎; 田上貴祥; 本同宏成; 森春英; 奥山正幸; 姚閔; 木村淳夫, 応用糖質科学, 1, 3, (37), 20 Jul. 2011
Japanese

GH family 31に属するAspergillus niger由来α‐glucosidaseの鎖長特異性の改変
田上貴祥; 奥山正幸; 森春英; 木村淳夫, 応用糖質科学, 1, 3, (36), 20 Jul. 2011
Japanese

Ruminococcus albus NE1株由来マンノシルグルコース加リン酸分解酵素の諸性質と一次構造の解析
川原良介; 伊藤重陽; 田口秀典; 佐分利亘; 森春英; 松井博和, 応用糖質科学, 1, 3, (51), 20 Jul. 2011
Japanese

イネ培養細胞を用いたスクロース誘導性タンパク質の網羅的解析
アンストーン ワスサン; 佐分利亘; 和久田真司; 濱田茂樹; 伊藤浩之; 森春英; 今井亮三; 松井博和, 応用糖質科学, 1, 3, (34), 20 Jul. 2011
Japanese

アノマー反転型トレハラーゼ変異体により合成された新規非還元二糖の構造
平内亨; 牧孝多朗; 森春英; 奥山正幸; 木村淳夫, 応用糖質科学, 1, 3, (36), 20 Jul. 2011
Japanese

Glycoside Hydrolase Family 63に属する酵素YgjKのグライコシンターゼ化
宮崎剛亜; 松田佳奈; 森春英; 北岡本光; 北野克和; 西河淳; 殿塚隆史, 応用糖質科学, 1, 3, (35), 20 Jul. 2011
Japanese

環状イソマルトオリゴ糖グルカノトランスフェラーゼのβ→αループ2はCI‐8の優先的生成に寄与する
齊藤みどり; KANG Hee‐Kwon; 森春英; 奥山正幸; 藤本瑞; 舟根和美; 小林幹彦; 木村淳夫, 応用糖質科学, 1, 3, (39), 20 Jul. 2011
Japanese

Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity
Young-Min Kim; Ryoko Shimizu; Hiroyuki Nakai; Haruhide Mori; Masayuki Okuyama; Min-Sun Kang; Zui Fujimoto; Kazumi Funane; Doman Kim; Atsuo Kimura, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 91, 2, 329, 339, Jul. 2011
English

Ruminococcus albus NE1のマンナン代謝の解析
中島碧; 伊藤重陽; 田口秀典; 佐分利亘; 森春英; 松井博和, 日本農芸化学会大会講演要旨集, 2011, 106, 05 Mar. 2011
Japanese

Ruminococcus albus NE1由来cellobiose phosphorylaseの一次構造の解析と大腸菌による組換え酵素の諸性質
阿部正太郎; 羽村健; 河内慎平; 伊藤重陽; 森本奈保喜; 佐分利亘; 森春英; 松井博和, 日本農芸化学会大会講演要旨集, 2011, 44, 05 Mar. 2011
Japanese

ホタテガイ(Pastinopecton yessoensis)中腸腺由来α‐glucosidase—Cl⁻による反応速度の上昇と基質特異性の変化—
桝田安志; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2011, 43, 05 Mar. 2011
Japanese

Dextran glucosidase(DexB)のウォーターパス改変酵素の機能解析
小林桃子; 本同宏成; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2011, 43, 05 Mar. 2011
Japanese

アノマー反転型トレハラーゼ変異体によるオリゴ糖合成
平内亨; 牧孝多朗; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2011, 44, 05 Mar. 2011
Japanese

環状イソマルトオリゴ糖グルカノトランスフェラーゼが示すCI‐8の優先的生成に寄与する構造因子
齋藤みどり; KANG Hee‐Kwon; 森春英; 奥山正幸; 藤本瑞; 舟根和美; 小林幹彦; 木村淳夫, 日本農芸化学会大会講演要旨集, 2011, 42, 05 Mar. 2011
Japanese

Ruminococcus albus NE1株由来cellodextrin phosphorylaseの酵素化学的諸性質
澤野達也; 佐分利亘; 森本奈保喜; 森春英; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2011, 14, 2011
Japanese

中等度好熱性アルカリ細菌Bacillus sp.AAH‐31株由来アミラーゼ遺伝子のクローニングおよび大腸菌組換え酵素の諸性質
向井惇; 金大勳; 森本奈保喜; 竹花稔彦; 佐分利亘; 森春英; 小池誠治; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2011, 14, 2011
Japanese

Rhodothermus marinus JCM9785由来耐熱性セロビオース2‐エピメラーゼのエピラクトースの効率的合成への応用
佐藤央基; 佐分利亘; 小島晃代; 田口秀典; 森春英; 松井博和, 日本農芸化学会北海道支部講演会講演要旨, 2011, 7, 2011
Japanese

Comparison of Enzymatic Properties and Gene Expression Profiles of Two Tuberonic Acid Glucoside β-Glucosidases from Oryza sativa L.
WAKUTA Shinji; HAMADA Shigeki; ITO Hiroyuki; IMAI Ryozo; MORI Haruhide; MATSUURA Hideyuki; NABETA Kensuke; MATSUI Hirokazu, Journal of applied glycoscience, 58, 2, 67, 70, 2011
Japanese Society of Applied Glycoscience, English

Oligo‐1,6‐glucosidaseがCl⁻依存性を示すために必要なアミノ酸残基の決定
寺田智明; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会東北支部大会プログラム・講演要旨集, 145th, 49, 27 Sep. 2010
Japanese

加水分解反応の抑制により縮合反応の生成物を蓄積させるグルコアミラーゼ変異体
砂守このみ; 森春英; 奥山正幸; 森本奈保喜; 松井博和; 木村淳夫, 日本農芸化学会東北支部大会プログラム・講演要旨集, 145th, 49, 27 Sep. 2010
Japanese

isomaltooligosaccharide6‐α‐glucosyltransferaseのサブサイト+1変異酵素の機能解析
西村崇志; 鐘ケ江倫世; 本同宏成; 奥山正幸; 森春英; 木村淳夫, J Appl Glycosci, 57, Suppl., 43, 20 Jul. 2010
Japanese

Aspergillus niger α‐glucosidaseのサブサイト+1の改変
田上貴祥; 奥山正幸; 森春英; 木村淳夫, J Appl Glycosci, 57, Suppl., 44, 20 Jul. 2010
Japanese

Oligo‐1,6‐glucosidaseの変異導入によるCl⁻イオン依存型への改変
寺田智明; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2010, 22, 05 Mar. 2010
Japanese

大腸菌組換えデキストリンデキストラナーゼのオリゴ糖に対する作用と触媒アミノ酸残基の解析
貞廣樹里; 佐分利亘; 森春英; 奥山正幸; 岡田嚴太郎; 木村淳夫, 日本農芸化学会大会講演要旨集, 2010, 22, 05 Mar. 2010
Japanese

GH family 31 α‐グルコシダーゼにおける基質の鎖長認識機構の解明
田上貴祥; 西村崇志; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2010, 22, 05 Mar. 2010
Japanese

Suicide Substrate-based Inactivation of Endodextranase by ω-Epoxyalkyl α-D-Glucopyranosides
KANG Hee-Kwon; KIM Young-Min; NAKAI Hiroyuki; KANG Min-Sun; HAKAMADA Wataru; OKUYAMA Masayuki; MORI Haruhide; NISHIO Toshiyuki; KIMURA Atsuo, Journal of Applied Glycoscience, 57, 4, 269, 277, 2010
Three kinds of ω-epoxyalkyl α-glucopyranosides (3′,4′-epoxybutyl α-D-glucopyranoside (E4G), 4′,5′-epoxypentyl α-D-glucopyranoside (E5G) and 5′,6′-epoxyhexyl α-D-glucopyranoside (E6G)), having alkyl chains of different lengths at their aglycone moieties, inactivated the endodextranase from Streptococcus mutans ATCC 25175 (SmDex) irreversibly with the pseudo-first order kinetics. Alkyl chain length-dependent inactivation was observed and the degree of activity loss was E5G, E6G and E4G, in that order, implying that the distance between epoxide group and glucosyl residue of ω-epoxyalkyl α-glucopyranoside was important in the modification of endodextranase. Inactivation by E5G followed the model of reversible intermediate-complex formation mechanism (suicide inhibitor-based mechanism). The rate constant of irreversible inactivation (k) and the dissociation constant of intermediate-complex (KR) of SmDex and E5G were 0.44 min^-1 and 1.45 mM, respectively. Hydrolytic reaction product (isomaltose) protected SmDex from E5G-inactivation, suggesting that E5G bound to the catalytic site of SmDex. This is the first report that ω-epoxyalkyl α-glucopyranoside becomes a suicide substrate for endodextranase., The Japanese Society of Applied Glycoscience, English

The first alpha-1,3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31
Min-Sun Kang; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura, BIOCHIMIE, 91, 11-12, 1434, 1442, Nov. 2009
English

Catalytic Reaction Mechanism Based on alpha-Secondary Deuterium Isotope Effects in Hydrolysis of Trehalose by European Honeybee Trehalase
Haruhide Mori; Jin-Ha Lee; Masayuki Okuyama; Mamoru Nishimoto; Masao Ohguchi; Doman Kim; Atsuo Kimura; Seiya Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 73, 11, 2466, 2473, Nov. 2009
English

Streptococcus mutans由来dextran glucosidaseの部位特異的変異導入による糖転移活性の改変(2)
中塚大地; 本同宏成; 大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 56, Suppl., 37, 20 Jul. 2009
Japanese

isomaltooligosaccharide6‐α‐glucosyltransferaseの糖転移反応に関わる構造因子
西村崇志; 鐘ケ江倫世; KLM Young‐Min; 本同宏成; 奥山正幸; 森春英; 木村淳夫, J Appl Glycosci, 56, Suppl., 37, 20 Jul. 2009
Japanese

テンサイα‐glucosidaseのサブサイト+2および+3の形成に重要なアミノ酸残基の決定
田上貴祥; 奥山正幸; 森春英; 田口和憲; 木村淳夫, J Appl Glycosci, 56, Suppl., 38, 20 Jul. 2009
Japanese

Bacteroides thetaiotaomicron由来SusBの触媒反応におけるCa²⁺の役割
吉田拓弥; 奥山正幸; 本同宏成; 銚閔; 森春英; 木村淳夫, J Appl Glycosci, 56, Suppl., 38, 20 Jul. 2009
Japanese

Streptococcus mutans由来dextran glucosidaseの部位特異的変異導入による糖転移活性の改変(1)
本同宏成; 大塚博昭; 中塚大地; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 56, Suppl., 37, 33, 20 Jul. 2009
The Japanese Society of Applied Glycoscience, Japanese

Structural Comparison of Streptococcus mutans Dextran Glucosidase with Glucoside Hydrolases in GH13
HONDOH Hironori; OTSUKA RACHI Hiroaki; SABURI Wataru; MORI Haruhide; OKUYAMA Masayuki; KIMURA Atsuo, Journal of applied glycoscience, 56, 2, 111, 117, 20 Apr. 2009
In glycoside hydrorase family (GH) 13, α-glucosidase, oligo-1,6-glucosidase and dextran glucosidase, which hydrolyze the non-reducing end glucosidic linkages of maltooligo- and/or isomaltoolligosaccharides, are categorized as α-glucoside hydrolase. Despite a high similarity in the sequence and overall structure of those family enzymes, GH 13 α-glucoside hydrolases show a wide range of substrate specificity. Until now, three crystal structures of α-glucoside hydrolase, dextran glucosidase from Streptococcus mutans (DGase), oligo-1,6-glucosidase from Bacillus cereus (O16G), and α-glucosidase from Geobacillus sp. HTA-462 (GSJ) have been determined. In this study, we have performed the structural comparison of these α-glucoside hydrolases. Their overall structures are generally similar, and consist of three major domains A, B and C as found in many α-amylase family enzymes. The significant structural differences in these enzymes are mainly found in loop regions. GSJ has a shorter β→α loop 6 in a different orientation in addition to the disordered regions, whereas DGase and O16G show high similarity in their tertiary structures. Though these enzymes have the different substrate preference, they all possess the completely conserved configuration at subsite -1. Therefore, the substrate preference will be originated from the structure at subsite for the reducing end side of substrate. The substrate binding modes of these glucoside hydrolases were predicted by superimposing the substrate molecule of substrate-complex structures of DGase and α-amylase. DGase and O16G are thought to have a similar manner in substrate binding with conserved amino acid residues. The substrate recognition of GSJ at subsite -1 and +1 would be similar to that of α-amylases since the key residues in substrate binding are conserved in both primary and tertiary structures., The Japanese Society of Applied Glycoscience, English

Isomal tooligosaccharide 6‐α‐glucosyltransferase(I6GT)のイソマルトオリゴ糖への作用と転移に関する構造因子
西村崇志; 鐘ケ江倫世; KIM Young‐Min; 本同宏成; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2009, 41, 05 Mar. 2009
Japanese

アズキα‐アミラーゼ(VAA)のMet398は過酸化水素により特異的に酸化される
田中良幸; 森春英; 河合正悟; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2009, 44, 05 Mar. 2009
Japanese

加水分解反応の抑制によりトレハロースを縮合蓄積させるトレハラーゼ変異酵素
牧孝多朗; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2009, 41, 05 Mar. 2009
Japanese

Preliminary Structure Determination of Polysaccharide Extracted from Rhizobium sp.
Wongchawalit Jintanart; Hashidoko Yasuyuki; Okuyama Masayuki; Mori Haruhide; Chiba Seiya; Kimura Atsuo, Journal of Applied Glycoscience Supplement, 2009, 0, 39, 39, 2009
The Japanese Society of Applied Glycoscience, Japanese

Structural Comparison of Streptococcus mutans Dextran Glucosidase with Glucoside Hydrolases in GH13
本同宏成; 大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 56, 2, 111-117 (J-STAGE), 117, 2009
In glycoside hydrorase family (GH) 13, α-glucosidase, oligo-1,6-glucosidase and dextran glucosidase, which hydrolyze the non-reducing end glucosidic linkages of maltooligo- and/or isomaltoolligosaccharides, are categorized as α-glucoside hydrolase. Despite a high similarity in the sequence and overall structure of those family enzymes, GH 13 α-glucoside hydrolases show a wide range of substrate specificity. Until now, three crystal structures of α-glucoside hydrolase, dextran glucosidase from Streptococcus mutans (DGase), oligo-1,6-glucosidase from Bacillus cereus (O16G), and α-glucosidase from Geobacillus sp. HTA-462 (GSJ) have been determined. In this study, we have performed the structural comparison of these α-glucoside hydrolases. Their overall structures are generally similar, and consist of three major domains A, B and C as found in many α-amylase family enzymes. The significant structural differences in these enzymes are mainly found in loop regions. GSJ has a shorter β→α loop 6 in a different orientation in addition to the disordered regions, whereas DGase and O16G show high similarity in their tertiary structures. Though these enzymes have the different substrate preference, they all possess the completely conserved configuration at subsite -1. Therefore, the substrate preference will be originated from the structure at subsite for the reducing end side of substrate. The substrate binding modes of these glucoside hydrolases were predicted by superimposing the substrate molecule of substrate-complex structures of DGase and α-amylase. DGase and O16G are thought to have a similar manner in substrate binding with conserved amino acid residues. The substrate recognition of GSJ at subsite -1 and +1 would be similar to that of α-amylases since the key residues in substrate binding are conserved in both primary and tertiary structures., The Japanese Society of Applied Glycoscience, English

Structural and Functional Analysis of a Glycoside Hydrolase Family 97 Enzyme from Bacteroides thetaiotaomicron
Momoyo Kitamura; Masayuki Okuyama; Fumiko Tanzawa; Haruhide Mori; Yu Kitago; Nobuhisa Watanabe; Atsuo Kimura; Isao Tanaka; Min Yao, JOURNAL OF BIOLOGICAL CHEMISTRY, 283, 52, 36328, 36337, Dec. 2008
English

Dextran glucosidaseの加水分解および糖転移反応における基質特異性の変換
本同宏成; 大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 55, Suppl., 62, 148, 20 Jul. 2008
The Japanese Society of Applied Glycoscience, Japanese

大腸菌trehalaseの触媒アミノ酸残基の決定とglycosynthase反応
森春英; 西塔沙織; 尾川陽; 牧孝多朗; 奥山正幸; 木村淳夫, J Appl Glycosci, 55, Suppl., 50, 103, 20 Jul. 2008
The Japanese Society of Applied Glycoscience, Japanese

α‐1,4グルコシド結合特異的なdextran glucosidase変異体の基質認識機構
本同宏成; 大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 55, Suppl., 49, 99, 20 Jul. 2008
The Japanese Society of Applied Glycoscience, Japanese

Glycoside hydrolase family97アノマー保持型酵素の触媒機構
奥山正幸; 姚閔; 本同宏成; 北村百世; 森春英; 田中勲; 木村淳夫, J Appl Glycosci, 55, Suppl., 59, 20 Jul. 2008
Japanese

デキストリンデキストラナーゼ遺伝子の異宿主発現および組換え酵素の解析
貞廣樹里; 佐分利亘; 森春英; 奥山正幸; 岡田嚴太郎; 木村淳夫, J Appl Glycosci, 55, Suppl., 49, 100, 20 Jul. 2008
The Japanese Society of Applied Glycoscience, Japanese

Bacillus sp.由来isomaltooligosaccharide6‐α‐glucosyltrasferase(I6GT)の転移反応に関与する構造因子
鐘ケ江倫世; KIM Young‐Min; 本同宏成; 奥山正幸; 森春英; 木村淳夫, J Appl Glycosci, 55, Suppl., 50, 20 Jul. 2008
Japanese

テンサイα‐glucosidaseのcDNAクローニングとpichia pastorisによる組換え酵素の生産
田上貴祥; 奥山正幸; 森春英; 田口和憲; 木村淳夫, J Appl Glycosci, 55, Suppl., 48, 20 Jul. 2008
Japanese

Substrate recognition mechanism of alpha-1,6-glucosidic linkage hydrolyzing enzyme, dextran glucosidase from Streptococcus mutans
Hironori Hondoh; Wataru Saburi; Haruhide Mori; Masayuki Okuyama; Toshitaka Nakada; Yoshiki Matsuura; Atsuo Kimura, JOURNAL OF MOLECULAR BIOLOGY, 378, 4, 913, 922, May 2008
English

Substrate Recognition of Escherichia coli YicI (α-Xylosidase)
OKUYAMA Masayuki; KANG Min-Sun; YAOI Katsuro; MITSUISHI Yasushi; MORI Haruhide; KIMURA Atsuo, Journal of applied glycoscience, 55, 2, 111, 118, 20 Apr. 2008
Glycoside hydrolase family 31 (GH 31) is one of the most intriguing glycoside hydrolase families. This family contains α-glucosidase, α-xylosidase, α-glucan lyase and isomaltosyltransferase. Escherichia coli YicI (α-xylosidase) is a representative enzyme of GH 31 because its biochemical and structural studies have been thoroughly carried out. YicI is a strict α-xylosidase, which rigidly recognizes α-xyloside at the non-reducing terminal end, even though its amino acid sequence apparently displays similarity with α-glucosidases. Phe277, Cys307, Trp345 and Lys414 at the subsite-1 are important for α-xylosidase activity. The mutant YicI enzymes, which possesses Ile307/Asp308 instead of Cys307/Phe308 and which has a shorter β→α loop 1 of (β/α)8 barrel in place of the original longer loop, respectively, possess α-glucosidase activity. In the transxylosylation of YicI, glucose, mannose and allose are able to act as acceptors, but galactose, talose and gulose never do, implying that equatorial OH-4 of the aldopyranose is crucial for acting as an acceptor. YicI transfers α-xylosyl moieties to a specific hydroxy group in the acceptor sugar (except fructopyranose) showing 1,6 regioselectivity, which is in agreement with the structural feature of the aglycone-biding site. Among the transxylosylation products of YicI, α-D-xylopyranosyl-(1→6)-D-mannopyranose, α-D-xylopyranosyl-(1→6)-D-fructofuranose, and α-D-xylopyranosyl-(1→3)-D-fructopyranose are novel sugars. α-D-Xylopyranosyl-(1→6)-D-mannopyranose and α-D-xylopyranosyl-(1→6)-D-fructofuranose have the ability to inhibit rat intestinal α-glucosidases., The Japanese Society of Applied Glycoscience, English

新しく見出されたイソマルトオリゴ糖不均化酵素の遺伝子クローニングと異種宿主発現
鐘ケ江倫世; KIM Young‐Min; 中井博之; 本同宏成; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2008, 191, 05 Mar. 2008
Japanese

アクセプタ結合部位への変異導入によるdextran glucosidaseの糖転移生成物の制御
本同宏成; 大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2008, 191, 05 Mar. 2008
Japanese

Schwanniomyces occidentalis由来GH31 α‐glucosidaseへの変異導入によるグルコシド結合選択性の変換
西村茉利子; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2008, 33, 05 Mar. 2008
Japanese

デキストリンデキストラナーゼの一次構造決定および組換え酵素の解析
貞廣樹里; 佐分利亘; 森春英; 奥山正幸; 岡田嚴太郎; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2008, 25, 2008
Japanese

新規酵素isomal tool igosaccharide6‐α‐glucosyl transferase(I6GT)の機能解析
鐘ケ江倫世; KIM Young‐Min; 本同宏成; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2008, 25, 2008
Japanese

Aglycone specificity of Escherichia coli alpha-xylosidase investigated by transxylosylation
Min-Sun Kang; Masayuki Okuyama; Katsuro Yaoi; Yasushi Mitsuishi; Young-Min Kim; Haruhide Mori; Doman Kim; Atsuo Kimura, FEBS JOURNAL, 274, 23, 6074, 6084, Dec. 2007
English

Function-unknown glycoside hydrolase family 31 proteins, mRNAs of which were expressed in rice ripening and germinating stages, are alpha-glucosidase and alpha-xylosidase
Hiroyuki Nakai; Shigeki Tanizawa; Tatsuya Ito; Koutarou Kamiya; Young-Min Kim; Takeshi Yamamoto; Kazuki Matsubara; Makoto Sakai; Hiroyuki Sato; Tokio Imbe; Masayuki Okuyama; Haruhide Mori; Yoshio Sano; Seiya Chiba; Atsuo Kimura, JOURNAL OF BIOCHEMISTRY, 142, 4, 491, 500, Oct. 2007
English

Molecular cloning of cDNA for trehalase from the European honeybee, Apis mellifera L., and its heterologous expression in Pichia pastoris
Jin-Ha Lee; Saori Saito; Haruhide Mori; Mamoru Nishimoto; Masayuki Okuyama; Doman Kim; Jintanart Wongchawalit; Atsuo Kimura; Seiya Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 9, 2256, 2265, Sep. 2007
English

Crystallization and preliminary X-ray analysis of Streptococcus mutans dextran glucosidase
Wataru Saburi; Hironori Hondoh; Hideaki Unno; Masayuki Okuyama; Haruhide Mori; Toshitaka Nakada; Yoshiki Matsuura; Atsuo Kimura, ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 63, Pt9, 774, 776, Sep. 2007
English

新規なイソマルトオリゴ糖生成酵素の単離と性質
鐘ケ江倫世; KIM Young‐Min; 中井博之; 奥山正幸; 森春英; 木村淳夫, J Appl Glycosci, 54, Suppl., 43, 20 Jul. 2007
Japanese

Streptococcus mutans由来dextran glucosidaseの保存領域IVに存在するAsnの機能解析
大塚博昭; 本同宏成; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 54, Suppl., 33, 20 Jul. 2007
Japanese

大腸菌Yicl(α‐xylosidase)の基質認識の解析
奥山正幸; KANG Min‐Sun; 矢追克郎; 矢追克郎; 三石安; 森春英; 木村淳夫, J Appl Glycosci, 54, Suppl., 55, 20 Jul. 2007
Japanese

ホタテガイ由来塩素イオン依存性α‐amylaseの構造と機能
飯塚貴久; 小林和之; 中井博之; 奥山正幸; 森春英; 奈良岡哲志; 千葉誠哉; 木村淳夫, J Appl Glycosci, 54, Suppl., 32, 20 Jul. 2007
Japanese

Molecular cloning of cDNAs and genes for three alpha-glucosidases from European honeybees, Apis mellifera L., and heterologous production of recombinant enzymes in Pichia pastoris
Mamoru Nishimoto; Haruhide Mori; Tsuneharu Moteki; Yukiko Takamura; Gaku Iwai; Yu Miyaguchi; Masayuki Okuyama; Jintanart Wongchawalit; Rudee Surarit; Jisnuson Svasti; Atsuo Kimura; Seiya Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 7, 1703, 1716, Jul. 2007
English

Streptococcus mutans由来dextran glucosidase(DGase)の基質特異性の変換:α‐1,4結合加水分解酵素への改変
大塚博昭; 佐分利亘; 本同宏成; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2007, 206, 05 Mar. 2007
Japanese

デキストラン関連酵素が有するデキストラン結合ドメインの機能解析
中井博之; 金泳民; 原口慶子; 奥山正幸; 森春英; 舟根和美; 小林幹彦; 木村淳夫, 日本農芸化学会大会講演要旨集, 2007, 65, 05 Mar. 2007
Japanese

B.thetaiotaomicron BT1871の求核触媒残基変異酵素の性質
奥山正幸; KANG Minsun; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2007, 206, 05 Mar. 2007
Japanese

ホタテガイ閉殻筋由来α‐glucosidaseのマルチプルフォームの構造解析
飯塚貴久; 中井博之; 奥山正幸; 森春英; 奈良岡哲志; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2007, 206, 05 Mar. 2007
Japanese

Streptococcus mutans由来Dextran Glucosidase M198Wの結晶構造解析
本同宏成; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2007, 36, 2007
Japanese

クロマルハナバチ(Bombus terrestris)α‐グルコシダーゼ:アロステリック酵素の性質解明と遺伝子発現
佐藤なつ子; 鳥羽瀬輝; 中井博之; 西本完; 森春英; 奥山正幸; 木村淳夫, 生化学, 3P-0212, 2007
Japanese

Multiple forms of alpha-glucosidase in rice seeds (Oryza sativa L., var Nipponbare)
Hiroyuki Nakai; Tatsuya Ito; Masatoshi Hayashi; Koutarou Kamiya; Takeshi Yamamoto; Kazuki Matsubara; Young-Min Kim; Wongchawalit Jintanart; Masayuki Okuyama; Haruhide Mori; Seiya Chiba; Yoshio Sano; Atsuo Kimura, BIOCHIMIE, 89, 1, 49, 62, Jan. 2007
English

Purification and characterization of alpha-glucosidase I from Japanese honeybee (Apis cerana japonica) and molecular cloning of its cDNA
Jintanart Wongchawalit; Takeshi Yamamoto; Hiroyuki Nakai; Young-Min Kim; Natsuko Sato; Mamoru Nishimoto; Masayuki Okuyama; Haruhide Mori; Osamu Saji; Chanpen Chanchao; Siriwat Wongsiri; Rudee Surarit; Jisnuson Svasti; Seiya Chiba; Atsuo Kimura, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70, 12, 2889, 2898, Dec. 2006
English

ホタテガイ閉殻筋に存在するイオン要求性α‐glucosidaseの精製とその諸性質
飯塚貴久; 中井博之; 奥山正幸; 森春英; 奈良岡哲志; 千葉誠哉; 木村淳夫, J Appl Glycosci, 53, Suppl., 30, 30 Aug. 2006
Japanese

Bacillus subtills 168S由来α‐glucosidase(YugT)の基質特異性に関わるアミノ酸残基の推定
大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, J Appl Glycosci, 53, Suppl., 30, 30 Aug. 2006
Japanese

Bacteroides thetaiotaomicron由来SusBの触媒残基解析
丹澤史子; 奥山正幸; 北村百世; 森春英; 田中勲; 木村淳夫, J Appl Glycosci, 53, Suppl., 33, 30 Aug. 2006
Japanese

Dextran glucosidaseのα‐1,6グルコシド結合認識機構
本同宏成; 佐分利亘; 奥山正幸; 森春英; 松浦良樹; 木村淳夫, J Appl Glycosci, 53, Suppl., 34, 30 Aug. 2006
Japanese

クロマルハナバチ,Bombus ignitus,α‐GlucosidaseアイソザイムIの長鎖基質認識に関わるアミノ酸残基の決定
佐藤なつ子; 中井博之; 森春英; 奥山正幸; 千葉誠哉; 木村淳夫, J Appl Glycosci, 53, Suppl., 31, 30 Aug. 2006
Japanese

GST融合タンパク質を用いたイネα‐glucosidaseの澱粉粒吸着および分解機構の解析
谷沢茂紀; 中井博之; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, J Appl Glycosci, 53, Suppl., 43, 30 Aug. 2006
Japanese

Structural elements to convert Escherichia coli alpha-xylosidase (YicI) into alpha-glucosidase
M Okuyama; A Kaneko; H Mori; S Chiba; A Kimura, FEBS LETTERS, 580, 11, 2707, 2711, May 2006
English

Structural elements in dextran glucosidase responsible for high specificity to long chain substrate
Wataru Saburi; Haruhide Mori; Saori Saito; Masayuki Okuyama; Atsuo Kimura, BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 1764, 4, 688, 698, Apr. 2006
English

Glycoside hydrolase family 97酵素BT1871の反応機構
奥山正幸; 北村百世; 森春英; 田中勲; 木村淳夫, 日本農芸化学会大会講演要旨集, 2006, 307, 05 Mar. 2006
Japanese

Bacillus subtilis168S由来α‐glucosidase YugTの機能解析
大塚博昭; 佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2006, 154, 05 Mar. 2006
Japanese

セイヨウミツバチα‐glucosidaseアイソザイムIIのβ→αループ8変異酵素の機能解析
佐藤なつ子; 中井博之; 森春英; 奥山正幸; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2006, 154, 05 Mar. 2006
Japanese

アズキα‐amylaseのデンプン粒吸着能はMet398改変により変化する
河合正悟; 浜井英礼; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2006, 154, 05 Mar. 2006
Japanese

Bacillus subtilis 168S由来β‐N‐acetylglucosaminidase(YbbD)の求核触媒残基を酸素酸化型システインに置換した変異酵素の性質
須賀原千佳; 佐分利亘; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2006, 306, 05 Mar. 2006
Japanese

Identification and Manipulation of Subsite Structure and Starch Granule Binding Site in Plant α-Amylase
MORI Haruhide, Journal of applied glycoscience, 53, 1, 51, 56, 20 Jan. 2006
In germinating plant seeds, α-amylases degrade starch accumulated in seeds, and that requires two functions: catalysis itself and starch granule binding ability. All plant α-amylases belong to the α-amylase family and share the same catalytic machinery as other members, but are different in extended subsite structure accommodating the non-reducing end side of substrate even with high affinity, particularly in subsite -6, shown in α-amylases of kidney bean as well as barley. Barley α-amylase isozyme 1 (AMY1) mutants introduced site-directed mutagenesis along the predicted substrate binding site and the recent crystal structure solved in complex with a substrate occupying subsite -1 to -7 revealed that amino acid residues situated in a shallow cleft extending between domain A and B were involved in the subsite formation. Although plant α-amylases possess no additional starch-binding domain as seen in several α-amylases from microorganisms, plant α-amylases examined acted on starch granules. The residue corresponding to "sugar tongs" Tyr380_AMY1 was proven to be involved in starch granule binding in adzuki bean α-amylase., The Japanese Society of Applied Glycoscience, Japanese

Purification and characterization of the hyper-glycosylated extracellular alpha-glucosidase from Schizosaccharomyces pombe
M Okuyama; Y Tanimoto; T Ito; A Anzai; H Mori; A Kimura; H Matsui; S Chiba, ENZYME AND MICROBIAL TECHNOLOGY, 37, 5, 472, 480, Oct. 2005
English

Glucoamylase originating from Schwanniomyces occidentalis is a typical alpha-glucosidase
F Sato; M Okuyama; H Nakai; H Mori; A Kimura; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 69, 10, 1905, 1913, Oct. 2005
English

Oligosaccharide binding to barley alpha-amylase 1
Robert, X; R Haser; H Mori; B Svensson; N Aghajari, JOURNAL OF BIOLOGICAL CHEMISTRY, 280, 38, 32968, 32978, Sep. 2005
English

植物α‐アミラーゼのデンプン結合部位の同定と改変に関する研究
森春英, 飯島記念食品科学振興財団年報, 2003, 90-95, Aug. 2005
Japanese

セイヨウミツバチα‐Glucosidase IIのV167N変異酵素は高いK_mおよびk₀を与え,マルハナバチα‐Glucosidase IIの性質を示した
佐藤なつ子; 中井博之; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, J Appl Glycosci, 52, Suppl., 24, 20 Jul. 2005
Japanese

Acetobacter capsulatum由来dextrin dextranaseの一次構造の解析
佐分利亘; 奥山正幸; 森春英; 岡田厳太郎; 木村淳夫, J Appl Glycosci, 52, Suppl., 26, 20 Jul. 2005
Japanese

—ホタテガイ中腸腺由来イオン依存性α‐glucosidase—
飯塚貴久; 福川太郎; 西岡謙吾; 中井博之; 奥山正幸; 森春英; 吉田孝; 千葉誠哉; 木村淳夫, J Appl Glycosci, 52, Suppl., 26, 20 Jul. 2005
Japanese

植物α‐GlucosidaseのC末端領域は単独で澱粉粒に吸着する‐GST融合タンパク質による解析‐
谷沢茂紀; 中井博之; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, J Appl Glycosci, 52, Suppl., 25, 20 Jul. 2005
Japanese

植物α‐Glucosidase,特にイネ酵素の分子解析と発芽時の澱粉粒分解に関する研究
中井博之; 谷沢茂紀; 松原一樹; 奥山正幸; 森春英; 千葉誠哉; 佐野芳雄; 木村淳夫, J Appl Glycosci, 52, Suppl., 52, 20 Jul. 2005
Japanese

Bacteroides thetaiotaomicron SusBの機能解析
奥山正幸; 北村百世; 丹沢史子; 北郷悠; 森春英; 田中勲; 木村淳夫, J Appl Glycosci, 52, Suppl., 25, 20 Jul. 2005
Japanese

Molecular analysis of alpha-glucosidase isozymes from Japanese honeybees (Apis cerana)
J Wongchawalit; T Yamamoto; M Okuyama; H Mori; R Surarit; J Svasti; S Chiba; AK Kimura, FEBS JOURNAL, 272, 101, 101, Jul. 2005
English, Summary international conference

Bacteroides thetaiomicron VPI‐5482由来Glycoside hydrolase family 66様タンパク質の発現と精製および機能解析
山本英治; 金泳民; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 196, 05 Mar. 2005
Japanese

Pichia pastorisを用いたアズキα‐Amylase組換酵素の高生産条件の検討
河合正悟; 浜井英礼; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 195, 05 Mar. 2005
Japanese

Glycoside hydrolase family 31酵素の構造と機能
奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 30, 05 Mar. 2005
Japanese

イネGlycoside Hydrolase Family 31様遺伝子の発現解析
中井博之; 伊藤真吾; 奥山正幸; 森春英; 千葉誠哉; 佐藤芳雄; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 30, 05 Mar. 2005
Japanese

クロマルハナバチ,Bombus ignitus,α‐Glucosidaseアイソザイムの精製・諸性質および一次構造の解析
佐藤なつ子; 高橋有志; 中井博之; 光畑雅宏; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 30, 05 Mar. 2005
Japanese

イネGlycoside hydrolase family 31様タンパク質の機能解析
谷沢茂紀; 中井博之; 奥山正幸; 森春英; 千葉誠哉; 佐野芳雄; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 31, 05 Mar. 2005
Japanese

ミツバチ由来結合型トレハラーゼのcDNAクローニングと組換え酵素の生産
西塔沙織; LEE Jin‐Ha; 西本完; 森春英; 奥山正幸; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 196, 05 Mar. 2005
Japanese

Bacillus subtilis 168S由来YbbDはβ‐N‐acetylglucosaminidase活性を示す。
須賀原千佳; 佐分利亘; 奥山正幸; 森春英; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 196, 05 Mar. 2005
Japanese

Streptococus mutans由来dextran glucosidaseの基質特異性の改変
佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 26, 05 Mar. 2005
Japanese

ミツバチα‐グルコシダーゼIIIの蜂蜜生産への適応:Tyr210の高K_nへの寄与
岩井岳; 森春英; 佐分利亘; 奥山正幸; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2005, 30, 05 Mar. 2005
Japanese

Involvement of individual subsites and secondary substrate binding sites in multiple attack on amylose by barley alpha-amylase
B Kramhoft; KS Bak-Jensen; H Mori; N Juge; J Nohr; B Svensson, BIOCHEMISTRY, 44, 6, 1824, 1832, Feb. 2005
English

Crystallization and preliminary X-ray analysis of alpha-xylosidase from Escherichia coli
M Kitamura; T Ose; M Okuyama; H Watanabe; M Yao; H Mori; A Kimura; Tanaka, I, ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 61, Pt 2, 178, 179, Feb. 2005
English

大腸菌Trehalase(TreA)の触媒残基の部位特異的変異導入による解析
西塔沙織; 森春英; 奥山正幸; 千葉誠哉; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2005, 16, 2005
Japanese

Streptococcus mutans由来dextran glucosidaseのMet198変異酵素の機能
佐分利亘; 森春英; 大塚博昭; 岩井岳; 奥山正幸; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2005, 15, 2005
Japanese

Glycoside hydrolase family31酵素の活性中心に保存されているArg残基の役割
汐川由希子; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 2005, 16, 2005
Japanese

Enzymatic synthesis of alkyl alpha-2-deoxyglucosides by alkyl alcohol resistant alpha-glucosidase from Aspergillus niger
YM Kim; M Okuyama; H Mori; H Nakai; W Saburi; S Chiba; A Kimura, TETRAHEDRON-ASYMMETRY, 16, 2, 403, 409, Jan. 2005
English

Localization of alpha-glucosidases I, II, and III in organs of European honeybees, Apis mellifera L., and the origin of alpha-glucosidase in honey
M Kubota; M Tsuji; M Nishimoto; J Wongchawalit; M Okuyama; H Mori; H Matsui; R Surarit; J Svasti; A Kimura; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68, 11, 2346, 2352, Nov. 2004
English

Overexpression and characterization of two unknown proteins, YicI and YihQ, originated from Escherichia coli
M Okuyama; H Mori; S Chiba; A Kimura, PROTEIN EXPRESSION AND PURIFICATION, 37, 1, 170, 179, Sep. 2004
English

Purification and characterization of Acremonium implicatum alpha-glucosidase having regioselectivity for alpha-1,3-glucosidic linkage
T Yamamoto; T Unno; Y Watanabe; M Yamamoto; M Okuyama; H Mori; S Chiba; A Kimura, BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 1700, 2, 189, 198, Aug. 2004
English

植物α‐Glucosidaseの生澱粉吸着に関与するアミノ酸残基の解析
中井博之; 谷沢茂紀; 奥山正幸; 森春英; 山本健; 佐野芳雄; 千葉誠哉; 木村淳夫, J Appl Glycosci, 51, Suppl., 41, 20 Jul. 2004
Japanese

アズキα‐アミラーゼの澱粉粒結合部位に関する研究
森春英; 浜井英礼; MAR S S; 千葉誠哉; 木村淳夫, J Appl Glycosci, 51, Suppl., 42, 20 Jul. 2004
Japanese

大腸菌yicl産物の機能およびX線構造解析
奥山正幸; 尾瀬農之; 北村百世; 森春英; 千葉誠哉; 木村淳夫; 田中勲, J Appl Glycosci, 51, Suppl., 40, 20 Jul. 2004
Japanese

クロマルハナバチ,Bombus ignitus,α‐Glucosidase Iの精製と諸性質
佐藤なつ子; 高橋有志; 中井博之; 光畑雅宏; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, J Appl Glycosci, 51, Suppl., 40, 20 Jul. 2004
Japanese

Search for a mechanism by which barley .ALPHA.-amylase recongnizes the substrate.
MORI HARUHIDE, 化学と生物, 42, 3, 170-172, 172, 25 Mar. 2004
Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese

Streptomyces sp. No.35由来β‐glucosidase F₃の活性化現象
峰島希; 福田健二; 森春英; 奥山正幸; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 255, 05 Mar. 2004
Japanese

触媒残基をcysteineに置換したdextran glucosidaseは酸化処理により活性が回復する
佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 255, 05 Mar. 2004
Japanese

植物α‐Glucosidaseに見出された新規な生澱粉結合部位の解析
中井博之; 奥山正幸; 森春英; 山本健; 千葉誠哉; 佐野芳雄; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 254, 05 Mar. 2004
Japanese

アズキα‐Amylaseのデンプン粒に対する吸着能は翻訳後修飾によって低下する
浜井英礼; 森春英; SAN SAN M; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 108, 05 Mar. 2004
Japanese

Glycosidase様タンパク質のglycosyl fluorideに対する反応の解析
奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 257, 05 Mar. 2004
Japanese

ミツバチα‐glucosidase III触媒残基変異酵素D206Gはアノマー反転型反応を行う。
岩井岳; 坪野真子; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 255, 05 Mar. 2004
Japanese

Bacillus subtilis 168S^r由来α‐glucosidase Phe144の機能解析
八巻勉; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 255, 05 Mar. 2004
Japanese

オオムギα‐アミラーゼ1 Asn209変異酵素の機能解析
福原有信; 森春英; 奥山正幸; SVENSSON B; 木村淳夫, 日本農芸化学会大会講演要旨集, 2004, 108, 05 Mar. 2004
Japanese

α‐Glucosidase変異酵素がα‐Glycosynthase反応を触媒する
奥山正幸; 森春英; 渡辺琴美; 木村淳夫; 千葉誠哉, 日本農芸化学会誌, 77, 11, 1140, 1141, 01 Nov. 2003
Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese

Purification, characterization and molecular cloning of tyrosinase from the cephalopod mollusk, Illex argentinus
T Naraoka; H Uchisawa; H Mori; H Matsue; S Chiba; A Kimura, EUROPEAN JOURNAL OF BIOCHEMISTRY, 270, 19, 4026, 4038, Oct. 2003
English

Purification, characterization, and sequence analysis of two alpha-amylase isoforms from azuki bean, Vigna angularis, showing different affinity towards beta-cyclodextrin sepharose
SS Mar; H Mori; JH Lee; K Fukuda; W Saburi; A Fukuhara; M Okuyama; S Chiba; A Kimura, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 67, 5, 1080, 1093, May 2003
English

オオムギα‐アミラーゼ1(AMY1)の部位特異的変異による基質特異性の改変
福原有信; 森春英; 奥山正幸; SVENSSON B; 木村淳夫, 日本農芸化学会大会講演要旨集, 2003, 98, 05 Mar. 2003
Japanese

赤米および日本晴で発芽時に作用するα‐Glucosidaseの一次構造の解析
中井博之; 伊藤達也; 松原一樹; 奥山正幸; 森春英; 千葉誠哉; 佐野芳雄; 木村淳夫, 日本農芸化学会大会講演要旨集, 2003, 100, 05 Mar. 2003
Japanese

Streptococcus mutans由来Dextran glucosidase変異酵素の長鎖基質に対する作用
佐分利亘; 森春英; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集, 2003, 99, 05 Mar. 2003
Japanese

β‐Cyclodextrinに異なる親和力を示すアズキα‐amylaseアイソフォーム
森春英; SAN SAN M; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2003, 98, 05 Mar. 2003
Japanese

糸状菌Acremonium implicatum IF030538由来α‐Glucosidase遺伝子のクローニング
山本健; 大畑祐一郎; 海野剛裕; 小川浩一; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2003, 100, 05 Mar. 2003
Japanese

Evidence of Intramolecular Transglucosylation Catalyzed by an .ALPHA.-Glucosidase.
Son Mee; Mori Haruhide; Okuyama Masayuki; Kimura Atsuo; Chiba Seiya, Journal of Applied Glycoscience, 50, 1, 41, 44, 2003
The hydrolytic reaction of carbohydrate-hydrolase is essentially accompanied by a reverse reaction (the condensation reaction), meaning that only the substrate capable of being hydrolyzed is produced by the reverse reaction. Honeybee α-glucosidase I can't hydrolyze isomaltose, but is capable of hydrolyzing maltose, kojibiose and slightly nigerose. Nevertheless, the enzyme catalyzes the formation and accumulation of isomaltose from glucose together with α-glucobioses such as maltose, kojibiose and nigerose. This finding is in conflict with the data that the enzyme has no hydrolytic activity toward isomaltose. However, the conflict for the peculiar phenomenon on the reaction was rationally explained by the evidence that isomaltose might be formed by the intramolecular transglucosylation via other α-glucobioses that are easily produced from glucose by the condensation reaction. It is suggested that the usual transglycosylation of carbohydrate-hydrolase may be accompanied by an intramolecular transfer reaction., The Japanese Society of Applied Glycoscience, English

Barley alpha-amylase Met53 situated at the high-affinity subsite-2 belongs to a substrate binding motif in the beta ->alpha loop 2 of the catalytic (beta/alpha)(8)-barrel and is critical for activity and substrate specificity
H Mori; KS Bak-Jensen; B Svensson, EUROPEAN JOURNAL OF BIOCHEMISTRY, 269, 22, 5377, 5390, Nov. 2002
English

Identification of essential ionizable groups and evaluation of subsite affinities in the active site of beta-D-glucosidase F-1 from a Streptomyces sp.
K Fukuda; H Mori; M Okuyama; A Kimura; H Ozaki; M Yoneyama; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66, 10, 2060, 2067, Oct. 2002
English

alpha-glucosidase mutant catalyzes "alpha-glycosynthase"-type reaction
M Okuyama; H Mori; K Watanabe; A Kimura; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66, 4, 928, 933, Apr. 2002
English

α‐Glucosidase変異酵素はGlycosynthase反応を触媒する
奥山正幸; 森春英; 木村淳夫; 千葉誠哉, 日本農芸化学会大会講演要旨集, 2002, 127, 05 Mar. 2002
Japanese

Acetobacter capsulatum由来Dextrin Dextranaseの初期反応の解析とその測定法の確立
佐分利亘; 奥山正幸; 森春英; 岡田厳太郎; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2002, 35, 05 Mar. 2002
Japanese

Bacillus subtilis 168S^r α‐glucosidase組換え体の大腸菌での生産
森春英; 佐分利亘; 尾関理香; 水上裕紀子; 木村淳夫; 千葉誠哉, 日本農芸化学会大会講演要旨集, 2002, 126, 05 Mar. 2002
Japanese

Aspergillus niger α‐Galactosidase遺伝子のPichia pastorisでの発現
矢守美典; 石原啓吾; 奥山正幸; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2002, 265, 05 Mar. 2002
Japanese

赤米α‐Glucosidaseの限定分解とそれによる諸性質への影響
中井博之; 伊藤達也; 森春英; 千葉誠哉; 木村淳夫, 日本農芸化学会大会講演要旨集, 2002, 127, 05 Mar. 2002
Japanese

Kinetic Studies on Substrate Specificity and Active Site of beta-D-Glucosidase F1 from Streptomyces sp.
FUKUDA K; SHIRAKAWA K; MORI H; OKUYAMA M; KIMURA A; OZAKI H; YONEYAMA M; CHIBA S, J. Appl. Glycosci., 49, 3, 265, 272, 2002

Modulation of activity and substrate binding modes by mutation of single and double subsites+1/+2 and-5/-6 of barley alpha-amylase 1
H Mori; K Sass Bak-Jensen; TE Gottschalk; M Saddik Motawia; Damager, I; B Lindberg Moller; B Svensson, EUROPEAN JOURNAL OF BIOCHEMISTRY, 268, 24, 6545, 6558, Dec. 2001
English

Purification and identification of the essential ionizable groups of honeybee, Apis mellifera L., trehalase
JH Lee; M Tsuji; M Nakamura; M Nishimoto; M Okuyama; H Mori; A Kimura; H Matsui; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 65, 12, 2657, 2665, Dec. 2001
English

ビールの国でのオオムギα-アミラーゼ研究 デンマーク・カールスバーグ研究所に滞在して
森 春英, 化学と生物, 39, 9, 588, 591, 25 Sep. 2001
学会出版センタ-, Japanese

Purification and substrate specificity of honeybee, Apis mellifera L., alpha-glucosidase III
M Nishimoto; M Kubota; M Tsuji; H Mori; A Kimura; H Matsui; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 65, 7, 1610, 1616, Jul. 2001
English

Carboxyl group of residue Asp647 as possible proton donor in catalytic reaction of alpha-glucosidase from Schizosaccharomyces pombe
M Okuyama; A Okuno; N Shimizu; H Mori; A Kimura; S Chiba, EUROPEAN JOURNAL OF BIOCHEMISTRY, 268, 8, 2270, 2280, Apr. 2001
English

アイソトープ効果に基づいたアミラーゼの反応機構
十川詩帆; 西本完; 福士幸治; 森春英; 木村淳夫; 千葉誠哉, 日本農芸化学会誌, 75, 65, 05 Mar. 2001
Japanese

赤米α‐Glucosidaseの精製と諸性質
中井博之; 林正敏; 森春英; 木村淳夫; 千葉誠哉, 日本農芸化学会誌, 75, 65, 05 Mar. 2001
Japanese

Isolation and Sequence of Putative alpha-Glucosidase Gene from Brevibacterium fuscum var. dextranlyticum Strain 0407
水野 隆文; 森 春英; 西本 完; 伊藤 浩之; 松井 博和; 木村 淳夫; 本間 守; 千葉 誠哉, J Appl Glycosci, 48, 3, 287, 291, 2001

Protein engineering of substrate binding subsites in amylolytic enzymes.
H Mori; KS Bak-Jensen; B Svensson, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 220, U117, U117, Aug. 2000
English, Summary international conference

Molecular cloning of isomaltotrio-dextranase gene from Brevibacterium fuscum var. dextranlyticum strain 0407 and its expression in Escherichia coli
T Mizuno; H Mori; H Ito; H Matsui; A Kimura; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 63, 9, 1582, 1588, Sep. 1999
English

The engineering of specificity and stability in selected starch degrading enzymes
B Svensson; KS Bak-Jensen; H Mori; J Sauer; MT Jensen; B Kramhoft; TE Gottschalk; T Christensen; BW Sigurskjold; N Aghajari; R Haser; N Payre; S Cottaz; H Driguez, RECENT ADVANCES IN CARBOHYDRATE BIOENGINEERING, 246, 272, 281, 1999
English

Expression characteristic of an enzyme related to carbon and nitrogen balance in the leaves of rice plants.
NAKAJIMA MASAKAZU; OSAKI MITSURU; SHINANO TAKURO; MORI HARUHIDE; TANNO TOSHIAKI, 日本土壌肥料学会講演要旨集, 44, 256, 256, Mar. 1998
一般社団法人日本土壌肥料学会, Japanese

Analysis of amino acid residue concerning catalytic activity of .ALPHA.-glucosidase in Schizosaccharomyces pombe.
OKUYAMA MASAYUKI; MORI HARUHIDE; KIMURA ATSUO; CHIBA SEIYA, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 1998, 15, 1998
Japanese

Localization of α-Glucosidase in Yeast Cells
Oyo Toshitsu Kagaku (J. Appl. Glycosci.), 45, 3, 281, 283, 1998

Molecular Cloning of an α-Amylase cDNA from Germinating Cotyledons of Kidney Bean (Phaseolus vulgaris L. cv. Toramame)
MORI Haruhide; KOBAYASHI Tetsuya; TONOKAWA Takashi; TATEMATSU Ayumi; MATSUI Hirokazu; KIMURA Atsuo; CHIBA Seiya, Journal of applied glycoscience, 45, 3, 261, 267, 1998
日本応用糖質科学会, English

A catalytic amino acid and primary structure of active site in Aspergillus niger alpha-glucosidase
A Kimura; M Takata; Y Fukushi; H Mori; H Matsui; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 7, 1091, 1098, Jul. 1997
English

Identification of essential ionizable groups in active site of Aspergillus niger alpha-glucosidase
A Kimura; A Somoto; H Mori; O Sakai; H Matsui; S Chiba, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 3, 475, 479, Mar. 1997
English

Brevibacterium fuscum. Cloning and analysis of primary structure.of var. dextranlyticum derived isomaltotrio-dextranase gene
MIZUNO TAKAFUMI; MATSUI HIROKAZU; MORI HARUHIDE; ITO HIROYUKI; KIMURA ATSUO; CHIBA SEIYA, 日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨, 1997, 6, 1997
Japanese

Molecular cloning and nucleotide sequences of cDNA and gene encoding endo-inulinase from Penicillium purpurogenum
S Onodera; T Murakami; H Ito; H Mori; H Matsui; M Honma; S Chiba; N Shiomi, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 60, 11, 1780, 1785, Nov. 1996
English

Amino acid sequence analysis of the active center vicinity of buckwheat seed .ALPHA.-glucosidase.
MORI ISSEI; IWANAMI SHUNSUKE; MORI HARUHIDE; KIMURA ATSUO; MATSUI HIROKAZU; CHIBA MASAYA, 日本農芸化学会東北支部講演要旨, 1996, Godo Gakujutsu Koenkai, 18, Sep. 1996
Japanese

Schizosaccharomyces pombe. Inference of the primary structure and active free radical of .ALPHA.-glucosidase.
OKUYAMA MASAYUKI; MORI HARUHIDE; KIMURA ATSUO; CHIBA MASAYA, 日本農芸化学会東北支部講演要旨, 1996, Godo Gakujutsu Koenkai, 17, Sep. 1996
Japanese

テンサイα-Glucosidaseの一次構造について : 酵素
岩波 俊介; 松井 博和; 伊藤 浩之; 木村 淳夫; 森 春英; 本間 守; 千葉 誠哉, 日本農藝化學會誌, 70, 138, 138, 05 Mar. 1996
社団法人日本農芸化学会, Japanese

トラマメ(Phaseolus vulgaris L.)の発芽子葉及び緑葉で発現するα-amylaseについて : 酵素
吉崎 成洋; 森 春英; 伊藤 浩之; 松井 博和; 千葉 誠哉, 日本農藝化學會誌, 70, 137, 137, 05 Mar. 1996
社団法人日本農芸化学会, Japanese

Schizosaccharomyces pombe. Estimation of primary structure and active dissociated group of .ALPHA.-glucosidase.
OKUYAMA MASAYUKI; MORI HARUHIDE; KIMURA ATSUO; CHIBA SEIYA, 日本農芸化学会北海道支部・北海道農芸化学協会シンポジウム及び合同学術講演会講演要旨, 1996, 17, 1996
Japanese

Amino acid sequence analysis near active center of buckwheat seed .ALPHA.-glucosidase.
MORI KAZUO; IWANAMI SHUNSUKE; MORI HARUHIDE; KIMURA ATSUO; MATSUI HIROKAZU; CHIBA SEIYA, 日本農芸化学会北海道支部・北海道農芸化学協会シンポジウム及び合同学術講演会講演要旨, 1996, 18, 1996
Japanese

Reinvestigation of Isomaltotrio-Dextranase from Brevibacterium fuscum var. dextranlyticum
MIZUNO Takafumi; MATSUI Hirokazu; ITO Hiroyuki; MORI Haruhide; KIMURA Atsuo; HONMA Mamoru; CHIBA Seiya, Journal of applied glycoscience, 43, 3, 347, 353, 1996
日本応用糖質科学会, English

植物α-Glucosidaseの活性中心 : 特にテンサイについて : 酵素
岩波 俊介; 伊藤 浩之; 木村 淳夫; 森 春英; 松井 博和; 本間 守; 千葉 誠哉, 日本農藝化學會誌, 69, 200, 200, 05 Jul. 1995
社団法人日本農芸化学会, Japanese

Aspergillus nigerの生産するα-galactosidaseに関する研究 : 酵素
伊木 繁雄; 谷本 佳博; 森 春英; 木村 淳夫; 松井 博和; 千葉 誠哉, 日本農藝化學會誌, 69, 198, 198, 05 Jul. 1995
社団法人日本農芸化学会, Japanese

Streptomyces sp. No.35菌体内β-glucosidaseの基質特異性 : 酵素
白川 康; 森 春英; 木村 淳夫; 松井 博和; 本間 守; 千葉 誠哉; 米山 道男; 尾崎 八郎, 日本農藝化學會誌, 69, 200, 200, 05 Jul. 1995
社団法人日本農芸化学会, Japanese

トラマメ(Phaseolus vulgaris L.)緑葉α-amylaseの精製と諸性質 : 酵素
吉崎 成洋; 森 春英; 松井 博和; 千葉 誠哉, 日本農藝化學會誌, 69, 12, 12, 05 Jul. 1995
社団法人日本農芸化学会, Japanese

CHEMICAL MODIFICATION AND AMINO-ACID-SEQUENCE OF ACTIVE-SITE IN SUGAR-BEET ALPHA-GLUCOSIDASE
S IWANAMI; H MATSUI; A KIMURA; H ITO; H MORI; M HONMA; S CHIBA, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 3, 459, 463, Mar. 1995
English

Substrate Specificity and Subsite Affinities of α-Amylase from Germinating Cotyledons of Phaseolus vulgaris L. cv Toramame.
Oyo Toshitsu Kagaku (J. Appl. Glycosci.), 42, 4, 387, 394, 1995

MOLECULAR-CLONING AND EXPRESSION OF AN ISOMALTO-DEXTRANASE GENE FROM ARTHROBACTER-GLOBIFORMIS T6
A IWAI; H ITO; T MIZUNO; H MORI; H MATSUI; M HONMA; G OKADA; S CHIBA, JOURNAL OF BACTERIOLOGY, 176, 24, 7730, 7734, Dec. 1994
English, Introduction scientific journal

テンサイα‐Glucosidaseの活性中心のアミノ酸配列
岩波俊介; 伊藤浩之; 木村淳夫; 松井博和; 森春英; 本間守; 千葉誠哉, 日本農芸化学会誌, 68, 3, 430, Mar. 1994
Japanese

Schizosaccharomyces pombe菌体外分泌α‐Glucosidaseの精製と諸性質
谷本佳博; 木村淳夫; 森春英; 松井博和; 千葉誠哉, 日本農芸化学会誌, 68, 3, 430, Mar. 1994
Japanese

Arthrobacter globiformis T6のIsomalto‐dextranase: 大腸菌による発現酵素の性質
水野隆文; 岩井淳; 伊藤浩之; 森春英; 松井博和; 本間守; 千葉誠哉; 岡田厳太郎, 日本農芸化学会誌, 68, 3, 427, Mar. 1994
Japanese

トラマメ(Phaseolus vulgaris L)緑葉に発現するα‐amylase
森春英; 小林哲也; 伊藤浩之; 松井博和; 本間守; 千葉誠哉, 日本農芸化学会誌, 68, 3, 428, Mar. 1994
Japanese

トラマメ(Phaseolus vulgaris L.)α‐amylase遺伝子の発現について
小林哲也; 森春英; 伊藤浩之; 松井博和; 本間守; 千葉誠哉, 日本農芸化学会誌, 68, 3, 428, Mar. 1994
Japanese

Penicillium purpurogenum由来endo‐inulinaseの遺伝子構造
邑上豊隆; 小野寺秀一; 塩見徳夫; 伊藤浩之; 森春英; 松井博和; 本間守; 千葉誠哉, 日本農芸化学会誌, 68, 3, 436, Mar. 1994
Japanese

Aspergillus nigerの生産するα‐galactosidaseに関する研究
伊木繁雄; 谷本佳博; 森春英; 松井博和; 千葉誠哉, 日本農芸化学会北海道支部・北海道農芸化学協会シンポジウム及び合同学術講演会講演要旨, 1994, 34, 1994
Japanese

STARCH-HYDROLYZING ENZYMES IN GERMINATING KIDNEY BEAN
H MORI; A TATEMATSU; H MATSUI; T TAKAYANAGI; M HONMA; S CHIBA, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 56, 9, 1499, 1500, Sep. 1992
English, Introduction scientific journal

応用微生物学特論, 2024年, 修士課程, 農学院

リーダーシップ学総論, 2024年, 修士課程, 農学院

大学院共通授業科目（一般科目）：自然科学・応用科学, 2024年, 修士課程, 大学院共通科目

食品安全・機能性開発学特論, 2024年, 修士課程, 農学院

食品安全・機能性開発学特論演習, 2024年, 修士課程, 農学院

一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), 2024年, 学士課程, 全学教育

化学概論, 2024年, 学士課程, 農学部

生物化学Ⅰ, 2024年, 学士課程, 農学部

生物化学Ⅱ, 2024年, 学士課程, 農学部

生物化学Ⅲ, 2024年, 学士課程, 農学部

Diversification of carbohydrate phosphorylases
Grants-in-Aid for Scientific Research
28 Jun. 2019 - 31 Mar. 2022
MORI Haruhide
To develop the enzymatic production of carbohydrates, phosphorylases were investigated in this study. A new activity, solabiose phosphorylase, was found. A possible metabolism involving the enzyme and a practical enzymatic synthesis of solabiose using this enzyme were shown.
Maltoside phosphorylase, as a member of starch-hydrolyzing enzyme family, was analysed intensively. Kinetic analysis revealed that the reaction followed the two-steps reaction, and phosphorolysis, transglycosylation, and hydrolysis occurred through competitive binding of the second substrates on the glycosyl enzyme intermediate. Significant change of the proportion of the three activities caused by mutations in possible binding residues suggested appropriate residues for the activities. Using the enzyme activity, insoluble α-glucans were produced though extension of their branched chains.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Research (Exploratory), Hokkaido University, 19K22263

糖質の多様化高機能化に向けた酵素法アプローチの新展開
科学研究費補助金(基盤研究(B))
Apr. 2018 - Mar. 2021
森 春英
文部科学省, Principal investigator, Competitive research funding

Survey on members of sucrose-world in Asian tropical reef, namely in Thailand
Grants-in-Aid for Scientific Research
2009 - 2012
KIMURA Atsuo; MORI Haruhide
Plants synthesize the starch in the leaves by photosynthesis. Leaf-starch is converted to sucrose, and transported to storage organ (e.g., tubers and fruit-bodies), followed by re-formation of starch (storage-starch). From many sites of bodies, plants secret sucrose, by which plants perform the cross-talk with environmental organisms (e.g., microorganisms and insects). Recently, it was found that huge amount of sucrose was secreted (this system is call as sucrose-world). The purpose of this research is survey of sucrose-world-members (environmental organisms and their enzymes) in Thailand, where high sunshine is available. As a result, the system of "sucrose -> polysaccharide -> secondary or ternary sugar" was investigated at sucrose secreted from roots. About flower bee, the enzyme converting sucrose to oligosaccharide was also investigated.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 21405020

1残基のアミノ酸置換で「糖質分解酵素を多糖合成酵素に変換」する現象の分子機構
科学研究費助成事業
2008 - 2009
木村 淳夫; 森 春英; 奥山 正幸
1残基のアミノ酸置換で「多糖の加水分解酵素(デキストラナーゼ)を合成酵素に転換できる現象」を見出した。この反応機構を分子解析することが、本申請の目的である。このような合成反応は例がなく、世界で初めての現象である。また、産業利用への発展にも期待したい。この残基は触媒アミノ酸と考えられる。本現象は試験管内の観察であるが、このような点突然変異した酵素が実際に生物で機能している可能性を得た。進化の過程においてアミノ酸置換は容易に生じ、1つのアミノ酸を変異させることで酵素分子を「加水分解→合成」にする戦略は、進化的に効率が良い。この戦略の検証も本申請の目的である。本年度は次の結果を得た。多糖合成の分子解析(デキストラナーゼ):1)酵素の結晶化と立体構造解析:昨年度に大量精製した親酵素とGly置換体を用いて結晶化条件を検討した。良好な結晶化条件を確定でき、X線構造解析を進行中である。2)他のアミノ酸による変異酵素:Asp→Gly置換体が合成反応を示したが、より効率の良いアミノ酸置換も想定されたため、他の残基への点変異を試みた。その結果、Gly置換体が最も高い反応効率を与えた。Glyは最もサイズの小さな残基であり、Asp→Gly置換で生じた大きな空間が重要と考えられた。すなわち、このサイズの大きい空間に陰イオンが侵入し合成反応が進行したと考えられた。3)陰イオンの解析:アザイドイオンが最も反応効率の良い陰イオンであった。従って本イオンのサイズ・強度が合成反応に最適であることが分かった。反応の至適pHを確定でき、pK_a値に大きな変化がないと予想できた。4)生成物の構造解析:生成多糖はデキストラン様の構造であった。触媒残基の変異酵素の解析(ウニ酵素):5)遺伝子の発現:酵素遺伝子の異種宿主発現を行った。酵素蛋白質は封入体を形成せず発現しているが、塩存在下であっても酵素活性が極めて低かった。
日本学術振興会, 挑戦的萌芽研究, 北海道大学, 20658025

Promotion of reverse reaction of carbohydrolase and its applicationfor oligosaccharide synthesis
Grants-in-Aid for Scientific Research(基盤研究(C))
2007 - 2008
Aruhide MORI
トレハラーゼはトレハロースを加水分解する.本研究では, トレハラーゼ改変酵素と特殊化合物(βフッ化グルコース)を用いて, 加水分解の逆反応によりトレハロースを高効率で合成させることに成功した.変異酵素として, 塩基触媒変異体, および塩基触媒および加水分解の基質の水分子に影響を与えるアミノ酸残基変異体を用いた.何れも反応速度は野生型に比べ低下したが, 特に, 後者の合成効率が高く, 60%程度の収率を示した.
Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 19580102

Molecular Analysis of Carbohydrases Showing Different Reaction by Novel Structure and Its Application
Grants-in-Aid for Scientific Research
2005 - 2007
KIMURA Atsuo; MORI Haruhide; OKUYAMA Masayuki
This project is about the enzymes having the structure similar to α-glucosidase; i.e. α-xylosidase, glucan lyase, cyclic-tetrasaccharide-forming enzyme, and α-glucosidase, each of which catalyzes the different reaction. Three-dimensional structure available recently allows us to analyze the molecular mechanism of reactions exhibited by four enzymes. The purposes of research are 1) to elucidate the relationship between substrate and amino acid residue(s) in the catalytic site; 2) to analyze the function of catalytic residues; 3) to elucidate the structural element(s) to display the above-described different reactions; 4) to synthesize the useful enzyme. Results are as follows.
(1) We analyzed the amino acid residues in the catalytic site of α-xylosidase to recognize α-xyloside-structure, and succeeded in conversion of α-xylosidase into α-glucosidase by the mutagenesis of its structural elements. (2) The catalytic residues were identified and their functions were investigated. (3) Amino acid replacement of α-glucosidase (a hydrolyzing enzyme) lost its hydrolytic activity and enhanced the transglucosidation ability, meaning the conversion of hydrolyzing enzyme into transferring enzyme. (4) We have succeed in change of transferring products.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 17380060

Synthesis of a-linked oligosaccharides by engineered enzymes
Grants-in-Aid for Scientific Research(基盤研究(C))
2005 - 2006
Haruhide MORI
The aim of this project was to establish new methods to synthesize alpha-linked oligosaccharides using engineered carbohydrate-hydrolysing enzymes. I focused the following two points : 1. establishment the new reaction suitable for oligosaccharide synthesis using catalytic residue-mutated "inactive" hydrolases. 2. changing specificities of the enzyme and screening enzymes showing novel specificities. In the point-1, a new basic method effective for oligosaccharide synthesis was developed, and through the point-2, wide varieties of oligosaccharide would be prepared.Point-1, catalytic residue...
Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 17580076

触媒アミノ酸を置換した酵素が、発揮する新たな機能とその応用
科学研究費助成事業
2004 - 2005
木村 淳夫; 森 春英; 奥山 正幸
糖質の加水分解酵素は、2つの酸性アミノ酸(AspやGlu)を触媒残基とし、それぞれが-COO^-と-COOHの荷電状態を形成し、協奏的に加水分解を触媒する。応用性の高い糖転移作用も示すが、分解と転移は2つの酸性アミノ酸でなされ分割できない。最近、α-グルコシダーゼにある-COO^-型の触媒基であるAspをCysに置換した。本酵素(Asp→Cys)には活性はないが、温和な酸化で活性を発揮した。Cysの-SHが酸化され-SOOHとなり、活性中心内で-SOO^-に解離し-COO^-の代わりを行うと考えている。この酵素は分解能を失い、糖転移能が上昇し95%の収率を与えた。本研究の目的は、-SOO^-酵素に見出された「非分解・高転移」の現象を解析することである。具体的には、1)酸化したCys残基の構造決定、2)糖転移反応の解析、3)他の酵素を合成酵素にする先駆けとして、触媒基を-SOO^-にした糖質酵素の構築と機帯解析、である。計画は順調に進行し、1)と2)が完了した。本年度は、この現象の応用を図るために3)の課題を中心に研究を進行させた。レバン合成酵素とキチン分解酵素を取り上げ、触媒残基をCysに置換し、酸化処理を行った。両酵素のCys変異体には活性がなかったが、穏やかな酸化により活性が回復した。導入したSH基が-SOOHに変化したことを確認した。Cys酸化酵素は、親酵素と異なる性質を示した(レバン合成酵素:至適pHや転移作用の変化、キチン分解酵素:至適pHや協同性の変化)。
日本学術振興会, 萌芽研究, 北海道大学, 16658136

Investigation on Molecular Mechanism of Sucrose-degrading Enzyme in Asian Honeybee.
Grants-in-Aid for Scientific Research
2004 - 2005
KIMURA Atsuo; MORI Haruhide; OKUYAMA Masayuki
We have analyzed three sucrose-degrading isozymes (α-glucosidase I,II, and III) in European honeybees, since these enzymes exhibited the interesting activity to sucrose of high concentration. α-Glucosidase I was a unique allosteric enzyme to display negative cooperativity in extremely high sucrose concentration. α-Glucosidase II also exhibited the allosteric property of positive cooperativity. α-Glucosidase III was a Michaelis-Menten-type enzyme to be secreted to nectar (bees gathered from flower) and to contribute to the formation of honey. Purpose of this project is i)existence of allosteric α-glucosidase in Asian honeybees, ii)investigation of honey-forming mechanism, and iii)regulation in expression of three α-glucosidase isozymes.
Four kinds of honeybee species were investigated in this research. From typical Asian honeybees (living in Thailand, Korea and Japan), three isozymes were isolated by chromatographic method. One of them was an allosteric enzyme having similar properties to European honeybee α-glucosidase I, allowing to molecular analysis of negative cooperativity. The second enzyme was α-glucosidase II-type isozyme. Enzyme in honey was also investigated, and showed similar to one of enzymes (α-glucosidase III-type isozyme). cDNAs of these three enzyme were cloned. It was found that the expression of three isozymes was regulated. In small honeybees living in Thailand, three α-glucosidases were also found, in which two α-glucosidases were isolated. Their properties investigated were almost identical to those of α-glucosidases I and II. Enzyme in honey was compared with adult bee enzymes. All honeybee species studied in this project contained allosteric enzyme. Asian honeybees were divided into two types : i)having three α-glucosidase isozymes and ii)having two α-glucosidase isozymes. Recently, we found that bumblebees also had two α-glucosidase isozymes. Further research will elucidate the interesting molecular mechanism of sucrose-degrading enzymes in flower bees.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), HOKKAIDO UNIVERSITY, 16405025

Research on Glycosylases which Obtained New Functions by Mutation on Catalytic Residue.
Grants-in-Aid for Scientific Research
2002 - 2004
KIMURA Atsuo; MORI Haruhide; OKUYAMA Masayuki
Glycosylases are enzymes that hydrolyze the glycosidic linkage. These enzymes also catalyze the transglycosidation, in which the glycosyl residue is transferred to the acceptor substrate. The transglycosidation is an important reaction i)to produce oligosaccharides valuable for foods and ii)to synthesize bio-active sugar-chains. Transglycosidation and hydrolysis proceed in the same time, meaning that the substrate for transglycosidation as well as its product(s) is cleaved by hydrolysis even under conditions of transglycosidation. We have studied the reactions of glycosylase, and have found the phenomena that catalyzed the transglycosidation only. In this study, we analyze the mechanism of valuable phenomena and perform their application. The results are summarized as follows. 1)We have determined the catalytic residue of negatively charged by the method using suicide substrate. Mutant enzyme (synthase), of which catalytic residue was replaced, was produce and purified. The enzyme showed no hydrolytic reaction, only catalyzed the synthesis of oligosaccharide(s) from fluoride-substrate and acceptor. Acceptor of aryl glycoside is a good substrate, meaning that the hydrophobic interaction between aryl-group and subsite +2 is important. 2)Mutant enzyme, which recognized the plane-shaped substrate, a mimic compound of reaction intermediate, was constructed, and its ability of oligosaccharide-synthesis was studied. The low production was observed. We changed the substrate concentration, and succeeded in the improvement of yield. Addition of alcohol to reaction mixture was also effective, but the high concentration of alcohol decreased the production of oligosaccharide. We have found a glycosidase resistant for alcohol. Currently, the conversion of alcohol-stable enzyme to mutant enzyme of same type is trying.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), HOKKAIDO UNIVERSITY, 14360043

Molecular Mechanism and Molecular Evolution of Glycosidases
Grants-in-Aid for Scientific Research(基盤研究(B))
2000 - 2002
千葉 誠哉; Haruhide MORI; 森 春英; 木村 淳夫; 福士 幸治
α-Glucosidase (EC 3.2.1.20), an exo-type glycosidase releasing α-glucose from substrate, is divided into two types of groups (family I and family II) that have different structures and substrate recognition. There are also the glycosidases, of which structures are homologous to each of family enzyme, such as α-amylase for family I and α-xylosidase for family II. The purpose of the project is to investigate the relationship of structure and function of two α-glucosidase families, and to understand the molecular mechanism of glycosidases. 1)The catalytic amino acid residues of two family enzy...
Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 北海道大学, Principal investigator, Competitive research funding, 12460035

活性中心の改変による新しいグリコシラーゼの作製
科学研究費助成事業
2001 - 2001
木村 淳夫; 森 春英
α-グルコシダーゼは、α-グルコシド結合をもつ基質に作用し、グルコースを遊離させる酵素である。転移反応では、α-グルコシル基を移し、有用な二糖類であるイソマルトースやニゲロースが工業的に生産されている。本酵素は、一次構造や基質認識が異なる2つのグループ(ファミリーIとII)に分類できる。触媒反応は、2つのカルボキシル基(_-COO^-と_-COOH)でなされ、この点では両ファミリーともに共通である。我々は、ファミリーI・II酵素の触媒残基(酸性アミノ酸)を自殺基質法や点突然変異法で決定した。II型酵素の_-COO^-である触媒基AspをAsn・Ala・Glyに置換した変異酵素は、加水分解反応を触媒できないが、活性中心の構造に大きな変化はない。最近、我々は触媒基AspのGly組換え酵素に糖転移活性が存在することを見い出した。本変異酵素は糖転移のみを一方的に行い、転移生成物を分解しない。本研究の目的は、新しく見い出されたこの現象を解析することであり、次に示す研究成果が得られた。(1)Asp→Ala酵素では本現象が認められず、Asp→Gly酵素のみが本反応を触媒した。Alaより小さなアミノ酸残基への置換が有効であった。(2)β-グルコシルフルオリドが第一基質となったが、α-グルコシルフルオリドでは反応が生じなかった。野生型酵素はα-型基質に作用するので、変異酵素の基質認識は逆転していた。従って、反応機構は縮合であると考えられた。(3)第二基質には、p-ニトロフェニル(PNPと略)α-グルコシド、α-キシロシド、α-マンノシドおよびβ-グルコシドが利用された。マルトースやPNPα-ガラクトシドには作用しなかった。PNPα-グルコシドの場合、約70%の高収率でPNPα-マルトシドとα-イソマルトシドが得られた。(4)現在、I型α-グルコシダーゼ、β-グルコシダーゼやα-ガラクトシダーゼについても本現象の解析を行っている。
日本学術振興会, 萌芽的研究, 北海道大学, 13876018

Application Studies on Product-Immobilization Phenomenon Newly Discovered
Grants-in-Aid for Scientific Research
1998 - 2000
KIMURA Atsuo; MORI Haruhide; CHIBA Seiya
Recently we have found the "immobilization of product" in the enzyme-digestion of starch granule, in which the product precipitates with granule. The phenomenon is of importance in i) the easy recovery of product under the less energy and ii) the inclusion of effective compounds into the polysaccharide. In this project, the fundamental study was done to learn the phenomenon, and the application of unutilized or low-utilized polysaccharides to the food industry was tried to be established. The results obtained were as follows.
(1) Cellulose. i) The product-immobilization was also found in the degradation of cellulose by cellulase. ii) Products in the cellulose of precipitate were water-soluble oligosaccharides of short-chain. iii) The inside product increased by addition of ethanol to reaction mixture. Saccharides in the cellulose was easily recovered under the low concentration of ethanol. iv) Production of oligosaccharides was done using ethanol. After enzyme reaction the supernatant was discarded, and then water was added to the precipitate, releasing oligosaccharides of different size. Isolation was done by gel-filtration and HPLC.(2) Chitin and xylan. i) In the enzymatic degradation, the product-immobilization was also found. However, the included amount was small as compared with starch granule and cellulose. ii) The addition of ethanol increased products in the two polysaccharides. (3) Porous starch granule as inclusion material. i) Porous starch granules of various plant origins were prepared by enzyme treatment. There were two types of starches which formed pores or no pore on the surface of granule. ii) The pore size could be controlled by amount and treatment time of enzyme. iii) The smallporous granule was able to incorporate the short-chain oligosaccharides, and the long-chain one entered the starch of large pore. iv) Ascorbic acid was also incorporated into the porous starch granule. After the granule including ascorbic acid was transferred into water, the compound gradually released into water.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B)., HOKKAIDO UNIVERSITY, 10556027

Molecular Mechanism of Novel N-Linked Sugar Chain Having α-Galactofuranosyl Structure
Grants-in-Aid for Scientific Research
1998 - 1999
KIMURA Atsuo; MORI Haruhide; CHIBA Seiya
In microorganisms there are the N- and O-linked sugar chains which are not found in animals and plants. Their biological function and synthesis have not been elucidated. In this project the structural determination of novel O-linked sugar chains was done, and the enzymes concerned with the formation of α-galactofuranosyl structure (Galf) in N-linked oligosaccharide, UDP-Galf synthetase and Galf transferase, were analyzed. (1) Five kinds of O-linked sugar chains, which were separated chemically from Aspergillus niger α-glu-cosidase, were purified, and the following structures were determined by monosaccharide analysis, exo-glycosidase treatment, and MS, 1D- and 2D-NMR: I) mannose, ii) mannobiose having α-1,2-linkage, iii) glucosylmannobiose of branched type, iv) two mannotrioses of branched (iv-a) and linear (iv-b) structures. Sugar chains of iii and iv-b were novel ones. (2) The substrate and product for Galf transferase were prepared from A .niger α-gluco-sidase. Since the activities of UDP-Galf synthetase and Galf transferase in cell extract of A .niger were low, the cultivation conditions were examined. The addition of maltose or starch to culture broth increased the both enzyme activities with induction of amylases (secretory proteins). When disruption of cells, the loss of activities was observed. It was found that the detergent stabilized the Galf transferase. The transferase preparation of high purity, which did not give a single band in electophoretic analysis, was obtained after several chromatographies. The purification is now doing to figure out the amino acid sequence and to separate the enzyme gene.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), HOKKAIDO UNIVERSITY, 10660068

Joint Studies on Dextran-Producing and -Degrading Enzymes
Grants-in-Aid for Scientific Research
1998 - 1999
CHIBA Seiya; MORI Haruhide; KIMURA Atsuo
Dextransucrase, one of glucansucrases, catalyzes the formation of dextran from sucrose. Recently Dr. Kim has established a novel mutation technique, Vacuum UV radiation. The mutant strains obtained hyper-produced dextransucrase constitutively, giving a possibility to produce the large amount of dextran. Dextran and its oligosaccharides have been found to have several useful functions to human. In the international project we tried to elucidate the relationship between structures and functions of dextran-producing and -degrading enzymes, and prepared the dextran and isomaltooligosaccharides by both enzymes.
(1) We succeeded in isolation of a hyper-produced dextransucrase gene and its expression in Escherichia coli. The mutated position was found in the promoter region. (2) A gene of isomaltotrio-dextranase was cloned and expressed in E. coli. The α-glucosidase gene was found in the upstream region of this gene. Both genes made a cluster structure, which was controlled by one promoter. (3) The large amount of dextran was prepared by hyper-produced dextransucrase. We examined the effective production method of pure isomaltotriose from dextran using isomaltotrio-dextranase. (4) An enzyme giving tetra- and penta-saccharides from dextran was purified and the properties were investigated. (5) Mechanism-based inactivator for dextranase was designed. Kinetic studies on inactivation indicated that compounds synthesized were found to be novel suicide substrate for dextranase.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B)., HOKKAIDO UNIVERSITY, 10044190

Structures and Substrate Recognitions of Animal, Botanical and Microbial α-Glucosidases and Their Molecular Evolution
Grants-in-Aid for Scientific Research
1997 - 1999
CHIBA Seiya; MORI Haruhide; KIMURA Atsuo
α-Glucosidase (EC 3.2.1.20) is a typical exo-type glycosidase that releases the α-glucose from non-reducing side of substrate. We are interested in the relationship between catalytic action and the structure, since the substrate specificity differs greatly with the source of enzyme. There are at least two types of α-glucosidases which show different substrate recognitions, suggesting that enzymes can be classified into two groups (family I and family II). However, the structural information is not enough to learn that structures of enzymes belonging to each group are homologous or not. In this project, we analyzed the primary structures of α-glucosidases from animal, botanical and microbial origins, and found that the enzymes of family I and II had different primary amino acid sequences.
α-Glucosidases from insect and bacteria belonged to family I, of which molecular weight was about 70 kDa. Four catalytic regions found were similar to those of α-amylase. The activity toward heteroside (sucrose or p-nitrophenyl α-glucoside) was higher than holoside (maltooligosaccharides). Members of family II were from animal, botanical and mold origins, and showed the opposite substrate specificity, high activity to holoside and low to heteroside. The molecular sizes were about 100 kDa. We analyzed the primary structures of six kinds of α-glucosidases belonging to this group. The sequences obtained were homologous each other, but no similarity was observed with family I enzymes. The findings suggest that the α-glucosidase was evolved from two different ancestral proteins.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 09460041

双子葉植物α-amylaseの細胞内局在性とそれに関与する因子の同定
科学研究費補助金(奨励研究(A))
1997 - 1998
森 春英
双子葉植物であるPhaseolus vulgarisにおいて、α-amylaseにアイソザイムは存在せず、発芽子葉と緑葉において同一の酵素が発現する。この同一のタンパク質が器官特異的な因子により異なる細胞内オルガネラに標的されている可能性を確認するために、まずPhaseolus vulgarisにおいてα-amylaseの細胞内局在性の確認と、発現制御の観点から遺伝子のクローニングを行った。(1) Phaseolus vulgarisにおける細胞内局在性の確認:Phaseolus vulgaris緑葉および発芽子葉における酵素の細胞内局在性を明らかにすることを目的に、ショ糖密度勾配法を用いた細胞内器官分画を行った。これにより、本酵素は子葉においてはプラスチド画分にあることが示唆され、一方緑葉においては少なくとも葉緑体画分に酵素は検出されなかった。さらに明確にするために、金コロイドによる免疫電子顕微鏡像を作製する。抗体を調製した。(2) Arabidopsisの形質転換(減圧浸潤法)をpBl121を用いて行った。また、pBl121上のGUSのN型糖鎖付加配列(-)の変異体GUS N356Sをレポーターに持つpBl124を創出した。これを用いて、本酵素上の器官特異的細胞内局在性因子を解析する。(3) α-Amylase遺伝子のクローニング:P.vulgans α-amylase遺...
文部科学省, 奨励研究(A), 北海道大学, Principal investigator, Competitive research funding, 09760062

植物の栄養ストレス下における糖代謝およびストレス関連酵素
科学研究費助成事業
1995 - 1995
松井 博和; 森 春英; 伊藤 浩之
本研究では、低栄養素ストレス下でも期待する収量が得られるよう、栄養素の利用効率を高めた作物の作出が急務と考え、作物が多量に集積するデンプンに着目した。サイトウやイネを材料に、デンプン合成・分解に関わる酵素と、いわゆるストレス関連酵素であるperoxidaseについて得られた成果を概説する。
1.デンプン枝付け酵素(Branching enzyme: BEと略)の精製と諸性質……トラマメ登熟種子よりBEを精製し、一般的な諸性質を明らかにした。本酵素はZn^<2+>やHg^<2+>などの金属イオンにより阻害されるばかりでなく、Ca^<2+>によっても強い阻害を受けた。N末端配列とプロテアーゼ消化による幾つかのペプチド断片の配列を解析したところ、イネおよびトウモロコシ起源のBEに高い相同性を示した。
2.α-Amylaseの精製とcDNAの解析……トラマメ発芽期と登熟期のcDNA解析を行った。両者には3'末端側のpolyA部分のみに相違が認められた。一方、緑葉中のα-Amylaseを単一に精製し、発芽種子α-Amylaseと性質や一次配列が酷似していることを明らかにした。
3.ストレス酵素Peroxidase遺伝子……イネperoxidaseをコードするprxRPN cDNAに対応する遺伝子(poxN)を単離し構造を解析した。さらに、プロモーター領域を5段階に削り込み、その下流にβ-グルクロニダーゼ(GUS)遺伝子を導入した各プラスミドを用いてタバコを形質転換し、得られた形質転換タバコ葉に幾つかの処理を施しGUS活性を測定した。
日本学術振興会, 重点領域研究, 北海道大学, 07263202

New Technique for High Yield Production of Glucose by Using of Mutarotase
Grants-in-Aid for Scientific Research
1993 - 1995
CHIBA Seiya; MORI Haruhide; ITO Hiroyuki; KIMURA Atsuo
(1) We kinetically analyzed the glucoamylase-catalyzed condensation of beta-glucose (two substrates reaction), and obtained the rate parameters and reaction rate equation which makes an accurate estimate of condensation product formation in any beta-glucose concentration. Addition of alpha-glucose to beta-glucose reaction system activated the rate of condensation. We analyzed this activation, and found that the subsite 1 of glucoamylase did not bind to alpha-glucose, meaning no inhibition to condensation, and that alpha-glucose had higher affinity to the subsite 2 than beta-glucose. We changed the composition of alpha-and beta-glucose with keeping the total glucose concentration constant, and measured the reaction rate. The velocity was reduced with decreasig in the molar fraction of beta-glucose, suggesting that the decrease of substrate (beta-glucose) is more effective than the activation by alpha-glucose and that the mutarotase suppresses the formation of condensation products.
(2) It was found that the mutarotase reduced the amount of disaccharides in 30% which was the products from 30% beta-glucose by glucoamylase. We investigated the effect of mutarotase on glucose production by two methods. The first was the butch-typed method which is presently used in the industrial glucose production system. The second was the immobilized enzyme technique where glucoamylase was linked to matrix, and then reaction was done in the column by running of maltodextrin with mutarotase. In both tests mutarotase gave the effective results, the increase of glucose production and the decrease of condensation products.
(3) We analyzed the amino acid sequence of porcine kidney mutarotase for developing the extensive project in the cloning of its gene and over-production of enzyme. The N-terminus of mutarotase was found to be blocked. We purified the N-terminal-blocked peptide fragment which was prepared by protease digestion, and determined its amino acid sequence by tandem mass spectrometry, elucidating that the acetyl group blocked the N-terminus of mutarotase.
Japan Society for the Promotion of Science, Grant-in-Aid for Developmental Scientific Research (B), HOKKAIDO UNIVERSITY, 05556011

植物の栄養ストレス下における糖代謝酵素の生化学・分子生物学的解析
科学研究費助成事業
1994 - 1994
松井 博和; 森 春英; 伊藤 浩之
低栄養条件下でも期待する収量が得られるよう、栄養素の利用効率を高めた作物を作出する観点から、本研究ではイネ科およびマメ科作物の栄養利用効率の差異を明らかにし、炭素および糖代謝化合物分配系、澱粉合成関連酵素ならびにストレス一般に関わる幾つかの酵素について、生化学・分子生物学的に解析することを目的とし、以下の成果を得た。
1。C-Nバランス酵素系:イネとダイズを標準培養液で水耕栽培し、種々の窒素条件下での呼吸速度を調べたところ、光呼吸および暗呼吸のいずれの速度をダイズの方が高かった。このような条件下における両作物のPEPCおよびSPS活性を測定したところ、窒素量とPEPC活性に相関が認められ、ダイズではいずれの活性もイネのそれら活性より低かったが、PEPC/SPSは高く、ダイズSPSはイネ酵素よりも窒素含有量に敏感には感応していないものと判断された。
2。初期光合成産物の同定:作物に^<14>CO_2を10分間吸収させ、その直後と30分後の葉を採取し、^<14>Cの分配を調べた。その結果、イネでは糖画分に多く分配されるのに対し、ダイズでは有機酸やアミノ酸に多く分配された。
3。澱粉合成時の酵素系:トラマメ登熟種子には少なくとも2種類のBranching Enzymeが存在することを明らかにした。DEAE-SepharoseおよびBio-Gel P-200を用いたクロマトグラフィーにより、その1つをSDS-PAGE的に単一に精製した。
4。ストレス酵素系:peroxidase isozymesの発現誘導機構を明らかにする目的で、その遺伝子断片を単離し、構造を解析した。また、5^1上流プロモーター領域の解析を、レポーターとしてβ-glucuronidase(GUS)遺伝子を用いた形質転換タバコで行った。
日本学術振興会, 重点領域研究, 北海道大学, 06271202

エピメリ化活性を有するタンパク質
Patent right, 佐分利亘; 森春英; 飯塚貴久; 藤本佳則; 高木宏基
特願2017-157664, 17 Aug. 2017
特開2019-033702, 07 Mar. 2019

α－１，６－グルコシル転移活性を有する酵素
Patent right, 森春英; 佐分利亘; 金井研太; 相沢健太; 飯塚貴久; 竹地紀昭; 谷美生夏
特願2018-028272, 22 Feb. 2018
特許6417061
31 Oct. 2018

マンノオリゴ糖合成酵素およびこれを用いたマンノオリゴ糖の製造法
Patent right, 森春英; 佐分利亘; 伊吹昌久; 津村和伸; 吉田靖彦
特願2017-032466, 23 Feb. 2017
特開2018-134058, 30 Aug. 2018