Ph.D., Hokkaido University

動物生化学

Glycosylation

Carbohydrate

X-ray crystallography

carbohydrate-active enzyme

molecular enzymology

Life Science, Functional biochemistry

Life Science, Applied biochemistry

Life Science, Animal life science

Bachelor's degree program, School of Agriculture

Master's degree program, Graduate School of Agriculture

Doctoral (PhD) degree program, Graduate School of Agriculture

Mar. 2023, 日本農芸化学会, 2023年度農芸化学奨励賞
糖質加水分解酵素の機能構造相関の解明と応用
田上貴祥

Mar. 2021, 日本農芸化学会, 第18回農芸化学研究企画賞
ブタ血液を原料とする酵素製剤の開発と糖尿病治療薬研究への応用
田上 貴祥

Feb. 2013, 日本応用糖質科学会北海道支部, 奨励賞
α-グルコシダーゼの長鎖マルトオリゴ糖特異性に関与する構造因子に関する研究
田上 貴祥

May 2011, 9th Carbohydrate Bioengineering Meeting, Best Poster Award
Takayoshi Tagami

Effects of osteopontin treatment alone and in combination with hormonal therapy on endometrial epidermal growth factor concentration and fertility at the first insemination after calving in dairy cows
Taiwo Kamardeen Bello; Takashi Tanida; Takayoshi Tagami; Yojiro Yanagawa; Seiji Katagiri
Theriogenology, 254, 117812, 117812, Elsevier BV, Apr. 2026, [Peer-reviewed]
English, Scientific journal

Porcine serum maltase-glucoamylase: structure, kinetics, and inhibition
Ken Watanabe; Takayoshi Tagami; Chihiro Biwa; Masato Kawasaki; Naruhiko Adachi; Toshio Moriya; Toshiya Senda; Masayuki Okuyama
Journal of Enzyme Inhibition and Medicinal Chemistry, 41, 1, 2612391, Informa UK Limited, 14 Jan. 2026, [Peer-reviewed], [Corresponding author]
English, Scientific journal

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, Jul. 2025, [Peer-reviewed], [Lead author, Corresponding author]
Scientific journal

Self-assembly of malto-oligosaccharide-block-solanesol in aqueous solutions: Investigating morphology and sugar-based physiological compatibility
Weeranuch Lang; Tomohisa Watanabe; Chaehun Lee; Sho Fukushima; Feng Li; Takuya Yamamoto; Kenji Tajima; Takayoshi Tagami; Redouane Borsali; Kenji Takahashi; Toshifumi Satoh; Takuya Isono
Carbohydrate Polymers, 123207, 123207, Elsevier BV, Mar. 2025, [Peer-reviewed]
Scientific journal

Fully biosourced amphiphilic block copolymer from tamarind seed xyloglucan and solanesol: synthesis, aqueous self-assembly, and drug encapsulation
Weeranuch Lang; Tomohisa Watanabe; Chaehun Lee; Takayoshi Tagami; Feng Li; Takuya Yamamoto; Kenji Tajima; Redouane Borsali; Kenji Takahashi; Toshifumi Satoh; Takuya Isono
Carbohydrate Polymers, 123181, 123181, Elsevier BV, Mar. 2025, [Peer-reviewed]
Scientific journal

Molecular structure of enzyme-synthesized amylose-like chimeric isomaltomegalosaccharides and their encapsulation of the sulfasalazine prodrug
Weeranuch Lang; Yoshiaki Yuguchi; Chun-Yao Ke; Ting-Wei Chang; Yuya Kumagai; Wilaiwan Kaenying; Takayoshi Tagami; Feng Li; Takuya Yamamoto; Kenji Tajima; Kenji Takahashi; Takuya Isono; Toshifumi Satoh; Atsuo Kimura
Carbohydrate Polymers, 122956, 122956, Elsevier BV, Feb. 2025, [Peer-reviewed]
English, Scientific journal

Identification of an osteopontin structural element for the restoration of a normal endometrial epidermal growth factor (EGF) profile determined by the EGF concentration on day 3 of estrous cycle and pregnancy outcome in repeat breeder dairy cows.
Takashi Tanida; Takayoshi Tagami; Yojiro Yanagawa; Seiji Katagiri
Theriogenology, 231, 171, 181, 01 Jan. 2025, [Peer-reviewed], [International Magazine]
English, Scientific journal, The loss of a cyclic change with two peaks of increased endometrial epidermal growth factor (EGF) concentration on days 2-4 and 13-14 during the estrous cycle has been linked to low fertility in repeat breeder (RB) cows. We have shown that an intravaginal infusion of osteopontin (OPN) restored the EGF profile in RB cows. The present study aimed to determine a structural element of OPN to restore the normal EGF profile and fertility. Holstein RB cows were diagnosed the EGF profile by a single examination of the endometrial EGF concentration on day 3 of the estrous cycle. Those with an altered EGF profile were intravaginally infused with OPN and its fragments on the day of insemination (day 0); the concentration of endometrial EGF was measured on day 3, and pregnancy was diagnosed on days 30-35. In Study 1, recombinant OPN (rOPN) (16 nmol), thrombin-cleaved N- and C-terminal fragments of rOPN (N-rOPN and C-rOPN, respectively), and a combination of these fragments (Th-rOPN) were infused (n = 13-20). The restoration rate of the normal EGF profile of the N-rOPN group (25.0 %) was a level in between the C-rOPN group (7.7 %) and both the rOPN (55.6 %) and Th-rOPN (64.3 %) groups. In Study 2, PBS (n = 47), rOPN (9.5 nmol, n = 83), and peptides of integrin binding motifs, GRGDSVAYGLK (peptide 1; 32, 320, and 1600 nmol), GRGDS (peptide 2; 320 and 1600 nmol), and SVAYGLK (peptide 3; 320 and 1600 nmol), were infused (n = 20-25). Restoration rates of the normal EGF profile of peptide 1 (320 and 1600 nmol) and peptide 3 (1600 nmol) groups (44.0-56.3 %) were comparable with those of the rOPN group (63.9 %) and higher than those of the PBS group (15.6 %). Restoration rates of the other groups were similar to those of the PBS group. Additional cows received infusions to determine the effect on fertility. Conception rates of the peptide 1 (320 and 1600 nmol; n = 50 each), peptide 3 (1600 nmol; n = 55), and rOPN (n = 111) groups (41.8-50.0 %) were comparable and higher than that of the PBS group (21.6 %, n = 75). In Study 3, PBS (n = 24), peptide 1 (320 nmol; n = 78), and GRGESVAYGLK peptide (peptide 4; 320 and 1600 nmol; n = 50 and 26, respectively) were infused. Restoration rates of the normal EGF profile of peptide 4 and PBS groups (16.0-19.2 %) were comparable and lower than those of the peptide 1 group (44.9 %). Thus, the SVAYGLK motif may be an OPN structural element to restore the normal EGF profile and fertility in RB cows, and the RGD motif may enhance its effect.

Mutant β-fructofuranosidase synthesizing blastose [β-d-Fruf-(2→6)-d-Glcp].
Atsuki Takagi; Takayoshi Tagami; Masayuki Okuyama
Enzyme and microbial technology, 180, 110500, 110500, Oct. 2024, [Peer-reviewed], [International Magazine]
English, Scientific journal, Fructooligosaccharides (FOS) are leading prebiotics that help keep the gut healthy and aid wellness by stimulating the growth and activity of beneficial intestinal bacteria. The best-studied FOS are inulin-type FOS, mainly oligosaccharides with β-Fruf-(2→1)-Fruf linkages, including 1-kestose [β-Fruf-(2→1)-β-Fruf-(2↔1)-α-Glcp] and nystose [β-Fruf-(2→1)-β-Fruf-(2→1)-β-Fruf-(2↔1)-α-Glcp]. However, the properties of other types of FOS-levan-type FOS with β-Fruf-(2→6)-Fruf linkages and neo-type FOS with β-Fruf-(2→6)-Glcp linkages-remain ambiguous because efficient methods have not been established for their synthesis. Here, using site-saturation mutation of residue His79 of β-fructofuranosidase from Zymomonas mobilis NBRC13756, we successfully obtained a mutant β-fructofuranosidase that specifically produces neo-type FOS. The H79G enzyme variant loses the native β-Fruf-(2→1)-Fru-transfer ability (which produces 1-kestose), and instead has β-Fruf-(2→6)-Glc-transfer ability and produces neokestose. Its hydrolytic activity specific to the β-Fruf-(2↔1)-α-Glcp bond of neokestose then yields blastose [β-Fruf-(2→6)-Glcp]. The enzyme produces 0.4 M blastose from 1.0 M sucrose (80 % of the theoretical yield). The production system for blastose established here will contribute to the elucidation of the physiological functions of this disaccharide.

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]
Scientific journal

Substrate Specificity of GH29 α-L-Glucosidases from Cecembia lonarensis.
Hye-Jin Kang; Takayoshi Tagami; Masayuki Okuyama
Journal of applied glycoscience, 71, 3, 91, 94, Aug. 2024, [Peer-reviewed], [Domestic magazines]
English, Scientific journal, We recently found two α-L-glucosidases, which can hydrolyze p-nitrophenyl α-L-glucopyranoside (PNP L-Glc) rather than p-nitrophenyl α-L-fucopyranoside, in glycoside hydrolase family 29. This study evaluated their substrate specificity for p-nitrophenyl α-L-rhamnopyranoside (PNP L-Rha), α-L-quinovopyranoside (PNP L-Qui), and α-L-xylopyranoside (PNP L-Xyl), of which structure is similar to PNP L-Glc. The two α-L-glucosidases had little activity toward PNP L-Rha. They exhibited higher k cat/K m values for PNP L-Qui but smaller for PNP L-Xyl than for PNP L-Glc. The molecular docking studies indicated that these specificities were correlated well with the active-site structure of the α-L-glucosidases. The finding that α-L-quinovoside, which has been suggested to occur in nature, is also a substrate for α-L-glucosidases indicates that this enzyme are not solely dedicated to α-L-glucoside hydrolysis.

Structural insights into starch-metabolizing enzymes and their applications
Takayoshi Tagami
Bioscience, Biotechnology, and Biochemistry, 88, 8, 864, 871, Oxford University Press (OUP), 28 May 2024, [Peer-reviewed], [Invited], [Lead author, Corresponding author]
English, Scientific journal, ABSTRACT

Starch is a polysaccharide produced exclusively through photosynthesis in plants and algae; however, is utilized as an energy source by most organisms, from microorganisms to higher organisms. In mammals and the germinating seeds of plants, starch is metabolized by simple hydrolysis pathways. Moreover, starch metabolic pathways via unique oligosaccharides have been discovered in some bacteria. Each organism has evolved enzymes responsible for starch metabolism that are diverse in their enzymatic properties. This review, focusing on eukaryotic α-glucosidases and bacterial α-glucoside-hydrolyzing enzymes, summarizes the structural aspects of starch-metabolizing enzymes belonging to glycoside hydrolase families 15, 31, and 77 and their application for oligosaccharide production.

Effects of recombinant osteopontin expressed in Escherichia coli on the recovery of the endometrial epidermal growth factor profile and fertility in repeat breeder dairy cows
Takashi Tanida; Takayoshi Tagami; Hiroko Sato; Hay Mar Kyaw; Takeshi Fujikawa; Masashi Nagano; Kenji Momozawa; Yojiro Yanagawa; Seiji Katagiri
Theriogenology, Elsevier BV, Jan. 2024, [Peer-reviewed]
Scientific journal

Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
Wilaiwan Kaenying; Takayoshi Tagami; Eukote Suwan; Chariwat Pitsanuwong; Sinchai Chomngam; Masayuki Okuyama; Palangpon Kongsaeree; Atsuo Kimura; Prachumporn T. Kongsaeree
Heliyon, 9, 11, e21923, e21923, Elsevier BV, Nov. 2023, [Peer-reviewed]
Scientific journal

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], [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.

Function and Structure of Lacticaseibacillus casei GH35 β-Galactosidase LBCZ_0230 with High Hydrolytic Activity to Lacto-N-biose I and Galacto-N-biose
Wataru Saburi; 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, 20 May 2023, [Peer-reviewed]
Scientific journal

Partial depolymerization of tamarind seed xyloglucan and its functionality toward enhancing the solubility of curcumin
Weeranuch Lang; Takayoshi Tagami; Hye-Jin Kang; Masayuki Okuyama; Nobuo Sakairi; Atsuo Kimura
Carbohydrate Polymers, 307, 120629, 120629, Elsevier BV, May 2023, [Peer-reviewed]
Scientific journal

Nonreducing terminal chimeric isomaltomegalosaccharide and its integration with azoreductase for the remediation of soil-contaminated lipophilic azo dyes
Weeranuch Lang; Sarote Sirisansaneeyakul; Takayoshi Tagami; Hye-Jin Kang; Masayuki Okuyama; Nobuo Sakairi; Atsuo Kimura
Carbohydrate Polymers, 305, 120565, 120565, Elsevier BV, Apr. 2023, [Peer-reviewed]
Scientific journal

Formulation and evaluation of a novel megalomeric microemulsion from tamarind seed xyloglucan-megalosaccharides for improved high-dose quercetin delivery
Weeranuch Lang; Debashish Mondol; Aphichat Trakooncharoenvit; Takayoshi Tagami; Masayuki Okuyama; Tohru Hira; Nobuo Sakairi; Atsuo Kimura
Food Hydrocolloids, 137, 108430, 108430, Elsevier BV, Apr. 2023, [Peer-reviewed]
Scientific journal

Crystal structure and identification of amino acid residues for catalysis and binding of GH3 AnBX β-xylosidase from Aspergillus niger
Wilaiwan Kaenying; Khuanjarat Choengpanya; Takayoshi Tagami; Pakorn Wattana-Amorn; Weeranuch Lang; Masayuki Okuyama; Yaw-Kuen Li; Atsuo Kimura; Prachumporn T. Kongsaeree
Applied Microbiology and Biotechnology, 107, 7-8, 2335, 2349, Springer Science and Business Media LLC, 06 Mar. 2023, [Peer-reviewed]
Scientific journal

Discovery of α-L-Glucosidase Raises the Possibility of α-L-Glucosides in Nature
Rikako Shishiuchi; Hyejin Kang; Takayoshi Tagami; Yoshitaka Ueda; Weeranuch Lang; Atsuo Kimura; Masayuki Okuyama
ACS Omega, 7, 50, 47411, 47423, American Chemical Society (ACS), 09 Dec. 2022, [Peer-reviewed], [Corresponding author]
Scientific journal

The location of protein oxidation in dystrophic skeletal muscle from the mdx mouse model of Duchenne muscular dystrophy
Tomohito Iwasaki; Jessica R. Terrill; Kei Kawarai; Yusei Miyata; Takayoshi Tagami; Naoyuki Maeda; Yasuhiro Hasegawa; Takafumi Watanabe; Miranda D. Grounds; Peter G. Arthur
Acta Histochemica, 124, 8, 151959, 151959, Elsevier BV, Dec. 2022, [Peer-reviewed]
Scientific journal

Physicochemical functionality of chimeric isomaltomegalosaccharides with α-(1 → 4)-glucosidic segments of various lengths
Weeranuch Lang; Yuya Kumagai; Shinji Habu; Juri Sadahiro; Takayoshi Tagami; Masayuki Okuyama; Shinichi Kitamura; Nobuo Sakairi; Atsuo Kimura
Carbohydrate Polymers, 291, 119562, 119562, Elsevier BV, Sep. 2022, [Peer-reviewed]
Scientific journal

Effects of milk osteopontin on the endometrial epidermal growth factor profile and restoration of fertility in repeat breeder dairy cows
Hay Mar Kyaw; Hiroko Sato; Takayoshi Tagami; Yojiro Yanagawa; Masashi Nagano; Seiji Katagiri
Theriogenology, 184, 26, 33, Elsevier BV, May 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Endometrial epidermal growth factor (EGF) shows a cyclic change with two peaks on Days 2-4 and 13-14 during the estrous cycle. An altered (i.e., loss of the two peaks) profile has been linked to reduced fertility in repeat breeder cows. We previously demonstrated that a form of osteopontin (OPN), with a molecular weight of 29 kDa and found in bull seminal plasma (SP), normalized the EGF profile and restored fertility in repeat breeder cows. OPN has many molecular forms due to post-translational modifications and is abundant in bovine milk. The purpose of the present study was to investigate whether mOPN normalizes the endometrial EGF profile and restores fertility in repeat breeder dairy cows with an altered EGF profile. OPN was separated by one-step anion-exchange column chromatography from the whey of bovine milk. Purified mOPN was verified by Western blotting and peptide mass fingerprinting analyses. The OPN fraction showed three major protein bands of 61, 37 and 31 kDa (peptides I, II, and III, respectively) on SDS-PAGE. All three major bands were identified as OPNs by Western blotting and their tryptic peptide masses were matched at approximately 50, 40, and 10%, respectively, to the bovine OPN amino acid sequence by a peptide mass finger printing analysis. The three bands accounted for approximately 85% of the total protein content and 6-23 mg of OPN was obtained from 1 L of bovine milk. A lyophilized eluate containing 1.3 mg of mOPN (171 cows), 0.5 mL of frozen SP (62 cows), and PBS (84 cows) was infused at estrus into the vagina of repeat breeder cows with an altered EGF profile. Some of the cows treated with mOPN, SP, and PBS (46, 50, and 45 cows, respectively) were inseminated immediately before the infusion and then examined for pregnancy between Days 60 and 65. The rate at which mOPN to normalize the EGF profile (56.1%) was similar to that of SP (58.1%) and higher than that of PBS (23.8%) (P < 0.05). The conception rate after the infusion of mOPN (43.5%) was similar to that of SP (40.0%) and higher than that of PBS (22.2%) (P < 0.05). The present results indicate that the infusion of mOPN into the vagina is a treatment option for repeat breeder cows with an altered EGF profile. Further studies are needed to compare the capacity of the three OPN molecules in milk to normalize the EGF profile, together with their molecular characteristics due to post-translational modifications.

Characterization of an Unknown Region Linked to the Glycoside Hydrolase Family 17 β-1,3-Glucanase of Vibrio vulnificus Reveals a Novel Glucan-Binding Domain
Yuya Kumagai; Hideki Kishimura; Weeranuch Lang; Takayoshi Tagami; Masayuki Okuyama; Atsuo Kimura
Marine Drugs, 20, 4, 250, 250, MDPI AG, 31 Mar. 2022, [Peer-reviewed]
English, Scientific journal, The glycoside hydrolase family 17 β-1,3-glucanase of Vibrio vulnificus (VvGH17) has two unknown regions in the N- and C-termini. Here, we characterized these domains by preparing mutant enzymes. VvGH17 demonstrated hydrolytic activity of β-(1→3)-glucan, mainly producing laminaribiose, but not of β-(1→3)/β-(1→4)-glucan. The C-terminal-truncated mutants (ΔC466 and ΔC441) showed decreased activity, approximately one-third of that of the WT, and ΔC415 lost almost all activity. An analysis using affinity gel containing laminarin or barley β-glucan revealed a shift in the mobility of the ΔC466, ΔC441, and ΔC415 mutants compared to the WT. Tryptophan residues showed a strong affinity for carbohydrates. Three of four point-mutations of the tryptophan in the C-terminus (W472A, W499A, and W542A) showed a reduction in binding ability to laminarin and barley β-glucan. The C-terminus was predicted to have a β-sandwich structure, and three tryptophan residues (Trp472, Trp499, and Trp542) constituted a putative substrate-binding cave. Linker and substrate-binding functions were assigned to the C-terminus. The N-terminal-truncated mutants also showed decreased activity. The WT formed a trimer, while the N-terminal truncations formed monomers, indicating that the N-terminus contributed to the multimeric form of VvGH17. The results of this study are useful for understanding the structure and the function of GH17 β-1,3-glucanases.

Structural insights reveal the second base catalyst of isomaltose glucohydrolase
Takayoshi Tagami; Minghao Chen; Yuta Furunaga; Asako Kikuchi; Juri Sadahiro; Weeranuch Lang; Masayuki Okuyama; Yoshikazu Tanaka; Tomohito Iwasaki; Min Yao; Atsuo Kimura
The FEBS Journal, 289, 4, 1118, 1134, Wiley, Feb. 2022, [Peer-reviewed], [Lead author, Corresponding author], [International Magazine]
English, Scientific journal, Glycoside hydrolase family 15 (GH15) inverting enzymes contain two glutamate residues functioning as a general acid catalyst and a general base catalyst, for isomaltose glucohydrolase (IGHase), Glu178 and Glu335, respectively. Generally, a two-catalytic residue-mediated reaction exhibits a typical bell-shaped pH-activity curve. However, IGHase is found to display atypical non-bell-shaped pH-kcat and pH-kcat /Km profiles, theoretically better-fitted to a three-catalytic residue-associated pH-activity curve. We determined the crystal structure of IGHase by the single-wavelength anomalous dispersion method using sulfur atoms and the cocrystal structure of a catalytic base mutant E335A with isomaltose. Although the activity of E335A was undetectable, the electron density observed in its active site pocket did not correspond to an isomaltose but a glycerol and a β-glucose, cryoprotectant, and hydrolysis product. Our structural and biochemical analyses of several mutant enzymes suggest that Tyr48 acts as a second catalytic base catalyst. Y48F mutant displayed almost equivalent specific activity to a catalytic acid mutant E178A. Tyr48, highly conserved in all GH15 members, is fixed by another Tyr residue in many GH15 enzymes; the latter Tyr is replaced by Phe290 in IGHase. The pH profile of F290Y mutant changed to a bell-shaped curve, suggesting that Phe290 is a key residue distinguishing Tyr48 of IGHase from other GH15 members. Furthermore, F290Y is found to accelerate the condensation of isomaltose from glucose by modifying a hydrogen-bonding network between Tyr290-Tyr48-Glu335. The present study indicates that the atypical Phe290 makes Tyr48 of IGHase unique among GH15 enzymes.

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, Springer Science and Business Media LLC, 13 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%.

Molecular insight into regioselectivity of transfructosylation catalyzed by GH68 levansucrase and β-fructofuranosidase
Masayuki Okuyama; Ryo Serizawa; Masanari Tanuma; Asako Kikuchi; Juri Sadahiro; Takayoshi Tagami; Weeranuch Lang; Atsuo Kimura
Journal of Biological Chemistry, 296, 100398, 100398, Elsevier BV, Jan. 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, Glycoside hydrolase family 68 (GH68) enzymes catalyze β-fructosyltransfer from sucrose to another sucrose, so-called transfructosylation. Although regioselectivity of transfructosylation is divergent in GH68 enzymes, there is insufficient information available on the structural factor(s) involved in the selectivity. Here, we found two GH68 enzymes, β-fructofuranosidase (FFZm) and levansucrase (LSZm), encoded tandemly in the genome of Zymomonas mobilis, displayed different selectivity: FFZm catalyzed the β-(2→1)-transfructosylation (1-TF), whereas LSZm did both of 1-TF and β-(2→6)-transfructosylation (6-TF). We identified His79FFZm and Ala343FFZm and their corresponding Asn84LSZm and Ser345LSZm respectively as the structural factors for those regioselectivities. LSZm with the respective substitution of FFZm-type His and Ala for its Asn84LSZm and Ser345LSZm (N84H/S345A-LSZm) lost 6-TF and enhanced 1-TF. Conversely, the LSZm-type replacement of His79FFZm and Ala343FFZm in FFZm (H79N/A343S-FFZm) almost lost 1-TF and acquired 6-TF. H79N/A343S-FFZm exhibited the selectivity like LSZm but did not produce the β-(2→6)-fructoside-linked levan and/or long levanooligosaccharides that LSZm did. We assumed Phe189LSZm to be a responsible residue for the elongation of levan chain in LSZm and mutated the corresponding Leu187FFZm in FFZm to Phe. An H79N/L187F/A343S-FFZm produced a higher quantity of long levanooligosaccharides than H79N/A343S-FFZm (or H79N-FFZm), although without levan formation, suggesting that LSZm has another structural factor for levan production. We also found that FFZm generated a sucrose analog, β-D-fructofuranosyl α-D-mannopyranoside, by β-fructosyltransfer to d-mannose and regarded His79FFZm and Ala343FFZm as key residues for this acceptor specificity. In summary, this study provides insight into the structural factors of regioselectivity and acceptor specificity in transfructosylation of GH68 enzymes.

Increased serum malondialdehyde concentration in cows with subclinical ketosis.
Senoh, T; Oikawa, S; Nakada, K; Tagami, T; Iwasaki, T
J. Vet. Med. Sci., 81, 817, 820, 2019, [Peer-reviewed]
English, Scientific journal

Novel α-1,3/α-1,4-glucosidase from Aspergillus niger exhibits unique transglucosylation to generate high levels of nigerose and kojibiose.
Ma, M; Okuyama, M; Tagami, T; Kikuchi, A; Klahan, P; Kimura, A
J. Agr. Food Chem., 67, 12, 2280, 2288, 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, α-Glucosidase from Aspergillus niger (AgdA; typical α-1,4-glucosidase) is known to industrially produce α-(1→6)-glucooligosaccharides. This fungus also has another α-glucosidase-like protein, AgdB. To learn its function, wild-type AgdB was expressed in Pichia pastoris. However, the enzyme displayed two electrophoretic forms due to heterogeneity of N-glycosylation at Asn354. The deglycosylation mutant N354D shared the same properties with wild-type AgdB. N354D demonstrated hydrolytic specificity toward α-(1→3)- and α-(1→4)-glucosidic linkages, indicating that AgdB is an α-1,3-/α-1,4-glucosidase. N354D-catalyzed transglucosylation from maltose was analyzed in short- and long-term reactions, enabling us to learn the transglucosylation specificity and product accumulation, respectively. A short-term reaction (<15 min) synthesized 3II- O-α-glucosyl-maltose and maltotriose, indicating α-1,3-/α-1,4-transferring specificity. A long-term reaction (<24 h) accumulated kojibiose and nigerose using formed glucose as an acceptor substrate. AgdA and AgdB are distinct α-glucosidases. At a high concentration of glucose added exogenously, AgdB largely generated the rare sugars kojibiose and nigerose (exhibiting beneficial physiological functions) with 19% and 24% yields from maltose, respectively.

Engineered dextranase from Streptococcus mutans enhances the production of longer isomaltooligosaccharides.
Klahan P; Okuyama M; Jinnai K; Ma M; Kikuchi A; Kumagai Y; Tagami T; Kimura A
Bioscience, Biotechnology, and Biochemistry, 82, 9, 1480, 1487, Sep. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Herein, we investigated enzymatic properties and reaction specificities of Streptococcus mutans dextranase, which hydrolyzes α-(1→6)-glucosidic linkages in dextran to produce isomaltooligosaccharides. Reaction specificities of wild-type dextranase and its mutant derivatives were examined using dextran and a series of enzymatically prepared p-nitrophenyl α-isomaltooligosaccharides. In experiments with 4-mg·mL-1 dextran, isomaltooligosaccharides with degrees of polymerization (DP) of 3 and 4 were present at the beginning of the reaction, and glucose and isomaltose were produced by the end of the reaction. Increased concentrations of the substrate dextran (40 mg·mL-1) yielded isomaltooligosaccharides with higher DP, and the mutations T558H, W279A/T563N, and W279F/T563N at the -3 and -4 subsites affected hydrolytic activities of the enzyme, likely reflecting decreases in substrate affinity at the -4 subsite. In particular, T558H increased the proportion of isomaltooligosaccharide with DP of 5 in hydrolysates following reactions with 4-mg·mL-1 dextran.Abbreviations CI: cycloisomaltooligosaccharide; CITase: CI glucanotransferase; CITase-Bc: CITase from Bacillus circulans T-3040; DP: degree of polymerization of glucose unit; GH: glycoside hydrolase family; GTF: glucansucrase; HPAEC-PAD: high performance anion-exchange chromatography-pulsed amperometric detection; IG: isomaltooligosaccharide; IGn: IG with DP of n (n, 2‒5); PNP: p-nitrophenol; PNP-Glc: p-nitrophenyl α-glucoside; PNP-IG: p-nitrophenyl isomaltooligosaccharide; PNP-IGn: PNP-IG with DP of n (n, 2‒6); SmDex: dextranase from Streptococcus mutans; SmDexTM: S. mutans ATCC25175 SmDex bearing Gln100‒Ile732.

A novel glycoside hydrolase family 97 enzyme: Bifunctional beta-L-arabinopyranosidase/alpha-galactosidase from Bacteroides thetaiotaomicron
Asako Kikuchi; Masayuki Okuyama; Koji Kato; Shohei Osaki; Min Ma; Yuya Kumagai; Kana Matsunaga; Patcharapa Kiahan; Takayoshi Tagami; Min Yao; Atsuo Kimura
BIOCHIMIE, 142, 41, 50, Nov. 2017, [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

Efficient synthesis of α-galactosyl oligosaccharides using a mutant Bacteroides thetaiotaomicron retaining α-galactosidase (BtGH97b)
Okuyama M; Matsunaga K; Watanabe K; Yamashita K; Tagami T; Kikuchi A; Ma M; Klahan P; Mori H; Yao M; Kimura A
The FEBS Journal, 284, 5, 766, 783, Feb. 2017, [Peer-reviewed], [International Magazine]
English, Scientific journal, The preparation of a glycosynthase, a catalytic nucleophile mutant of a glycosidase, is a well-established strategy for the effective synthesis of glycosidic linkages. However, glycosynthases derived from α-glycosidases can give poor yields of desired products because they require generally unstable β-glycosyl fluoride donors. Here, we investigate a transglycosylation catalyzed by a catalytic nucleophile mutant derived from a glycoside hydrolase family (GH) 97 α-galactosidase, using more stable β-galactosyl azide and α-galactosyl fluoride donors. The mutant enzyme catalyzes the glycosynthase reaction using β-galactosyl azide and α-galactosyl transfer from α-galactosyl fluoride with assistance of external anions. Formate was more effective at restoring transfer activity than azide. Kinetic analysis suggests that poor transglycosylation in the presence of the azide is because of low activity of the ternary complex between enzyme, β-galactosyl azide and acceptor. A three-dimensional structure of the mutant enzyme in complex with the transglycosylation product, β-lactosyl α-d-galactoside, was solved to elucidate the ligand-binding aspects of the α-galactosidase. Subtle differences at the β→α loops 1, 2 and 3 of the catalytic TIM barrel of the α-galactosidase from those of a homologous GH97 α-glucoside hydrolase seem to be involved in substrate recognitions. In particular, the Trp residues in β→α loop 1 have separate roles. Trp312 of the α-galactosidase appears to exclude the equatorial hydroxy group at C4 of glucosides, whereas the corresponding Trp residue in the α-glucoside hydrolase makes a hydrogen bond with this hydroxy group. The mechanism of α-galactoside recognition is conserved among GH27, 31, 36 and 97 α-galactosidases. DATABASE: The atomic coordinates (code: 5E1Q) have been deposited in the Protein Data Bank.

Effects of Collagen Casing-Containing Diets Fed to Sows during Late Pregnancy Lactation and Farrowing on the Birth Performance, Growth, Meat Product and Serum Components in Piglets
Michi YAMADA; Yuki WATANABE; Hanako YOSHIDA; Syunoa MITANI; Takayuki HARA; Takayoshi TAGAMI; Tomohito IWASAKI; Mikio SUGANO; Kazushige TAKEHANA; Shinji SUGIYAMA; Tetsuya EBIHARA; Yoh-ichi KOYAMA; Koji YAMADA; Hiroki NAKATSUJI
Nihon Yoton Gakkaishi, 54, 3, 130, 141, The Japanese Society of Swine Science, 2017, [Peer-reviewed]
Japanese, Scientific journal

Substrate recognition of the catalytic alpha-subunit of glucosidase II from Schizosaccharomyces pombe
Masayuki Okuyama; Masashi Miyamoto; Ichiro Matsuo; Shogo Iwamoto; Ryo Serizawa; Masanari Tanuma; Min Ma; Patcharapa Klahan; Yuya Kumagai; Takayoshi Tagami; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 81, 8, 1503, 1511, 2017, [Peer-reviewed]
English, Scientific journal

Functional characterization of UDP-rhamnose-dependent rhamnosyltransferase involved in anthocyanin modification, a key enzyme determining blue coloration in Lobelia erinus
Yang-Hsin Hsu; Takayoshi Tagami; Kana Matsunaga; Masayuki Okuyama; Takashi Suzuki; Naonobu Noda; Masahiko Suzuki; Hanako Shimura
PLANT JOURNAL, 89, 2, 325, 337, Jan. 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

Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1 -> 6)-alpha-nigerosyl
Takayoshi Tagami; Eri Miyano; Juri Sadahiro; Masayuki Okuyama; Tomohito Iwasaki; Atsuo Kimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 291, 32, 16438, 16447, Aug. 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

The loop structure of Actinomycete glycoside hydrolase family 5 mannanases governs substrate recognition
Yuya Kumagai; Keitaro Yamashita; Takayoshi Tagami; Misugi Uraji; Kun Wan; Masayuki Okuyama; Min Yao; Atsuo Kimura; Tadashi Hatanaka
FEBS JOURNAL, 282, 20, 4001, 4014, Oct. 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

Production of 1,5-anhydro-d-fructose by an alpha-glucosidase belonging to glycoside hydrolase family 31
Janjira Maneesan; Hideyuki Matsuura; Takayoshi Tagami; Haruhide Mori; Atsuo Kimura
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 78, 12, 2064, 2068, Dec. 2014, [Peer-reviewed]
English, Scientific journal

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

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

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

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

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

自然界にはα-L-グルコシド加水分解酵素が存在する α-L-グルコシダーゼの発見と構造・機能解析
奥山正幸; 田上貴祥, 化学と生物, 109, 111, Mar. 2024, [Peer-reviewed]
Japanese, Introduction scientific journal

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

リピートブリーダー牛にて、子宮内膜の上皮成長因子profileinを正常化する乳由来オステオポンチンの効果(Capacity of milk osteopontin to normalize endometrial epidermal growth factor profilein repeat breeder cows)
Kyaw Hay Mar; 田上 貴祥; 柳川 洋二郎; 永野 昌志; 片桐 成二, 日本獣医学会学術集会講演要旨集, 163回, 267, 267, Oct. 2020
(公社)日本獣医学会, English

組換えオステオポンチンによるリピートブリーダー牛の子宮内膜上皮成長因子(EGF)濃度正常化と受胎性回復効果の検証
佐藤 弘子; Hay Mar Kyaw; 柳川 洋二郎; 永野 昌志; 田上 貴祥; 片桐 成二, Journal of Mammalian Ova Research, 37, 1, S39, S39, Jul. 2020
(一社)日本卵子学会, Japanese

デキストランデキストリナーゼの多糖合成に関与するアミノ酸の同定
佐々木優希; 熊谷祐也; 貞廣樹里; LANG Weeranuch; 田上貴祥; 奥山正幸; 木村淳夫, 日本農芸化学会大会講演要旨集(Web), 2018, ROMBUNNO.2A26p03 (WEB ONLY), 05 Mar. 2018
Japanese

GH15 isomaltose glucohydrolaseの基質認識に関わる構造因子の同定
田上貴祥; 陳明皓; 奥山正幸; 岩崎智仁; 田中良和; 姚閔; 木村淳夫, 応用糖質科学, 7, 3, 49, 20 Aug. 2017
Japanese

放線菌Kribbella flavidaによる環状四糖の生成と代謝
田上 貴祥, Bioscience & Industry, 75, 2, 140, 141, Mar. 2017, [Invited]

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
田上 貴祥, 応用糖質科学, 6, 103, 108, 2016, [Peer-reviewed], [Invited]
Japanese, Introduction scientific journal

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

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

GH31 α-グルコシダーゼの基質特異性と構造の多様性
田上 貴祥, 化学と生物, 52, 205, 207, 2014, [Peer-reviewed], [Invited]
Japanese, Introduction scientific journal

Key aromatic residues at subsites +2 and +3 of glycoside hydrolase family 31 .ALPHA.-glucosidase contribute to recognition of long-chain substrates
TAGAMI Takayoshi; OKUYAMA Masayuki; NAKAI Hiroyuki; KIM Young-min; MORI Haruhide; TAGUCHI Kazunori; SVENSSON Birte; KIMURA Atsuo, Biochimica et Biophysica Acta, 1834, 1, 329, 335, Jan. 2013
English

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
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

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

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

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, 37, 20 Jul. 2009
The Japanese Society of Applied Glycoscience, 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, 94, 20 Jul. 2008
The Japanese Society of Applied Glycoscience, Japanese

Molecular basis for conversion of a glycoside hydrolase family 15 anomer-retaining α-glucosyltransferase to an anomer-inverting α-glucohydrolase
Teni QIN; Wataru SABURI; Shiho TAKEI; Takayoshi TAGAMI; Tomoya OTA; Kenta KANAI; Toyoyuki OSE; Haruhide MORI
日本農芸化学会2026年度大会, Oral presentation
09 Mar. 2026 - 12 Mar. 2026

縮合反応効率を低下させたグルコアミラーゼ変異酵素の開発
窪田大樹; 田上貴祥; 奥山正幸
日本農芸化学会2026年度大会, Oral presentation
09 Mar. 2026 - 12 Mar. 2026

Cecembia lonarensis LW9 株におけるL-glucose/L-quinovose 異化関連酵素の解析
赤津諒大; 田上貴祥; 奥山正幸
令和7年度日本応用糖質科学会北海道支部講演会, 19 Feb. 2026, Poster presentation

Neobacillus drentensis 57N由来イヌロスクラーゼ（NdIS）の結晶構造解析とイヌリン鎖長に影響を与える基質結合部位周辺のアミノ酸残基の同定
和田 歩; 佐分利 亘; 太田 智也; 武井 梓穂; 田上 貴祥; 城戸 悠輔; 尾瀬 農之; 森 春英
令和7年度日本応用糖質科学会北海道支部講演会, 19 Feb. 2026, Poster presentation

糖質加水分解酵素ファミリー15酵素の機能と構造の多様性
田上貴祥; 佐分利亘; 奥山正幸; 森 春英
第14回 応用糖質科学シンポジウム, 05 Sep. 2025, Japanese, Public symposium

α-L-グルコシダーゼホモログの基質特異性の解析
小川 真依; カン ヘジン; 田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

L-キノボースに結合特異性を示す新規solute-binding proteinの発見
龍井 草汰; 田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

GH97 α-グルコシド加水分解酵素触媒残基変異酵素の機能・構造解析
本郷 維規; 田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

マルターゼ-グルコアミラーゼに対するアカルビオシルマルトトリオースの阻害型式の解析
田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

イソマルトースグルコハイドロラーゼのグルコースおよびイソマルトースチオ類似体との複合体結晶構造解析
布施 朱理; 田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

Kribbella flavida 由来 GH15 トレハラーゼの機能と結晶構造の解析
眞鍋 佑輔; 田上 貴祥; 奥山 正幸
日本農芸化学会2025年度大会, 06 Mar. 2025, Japanese, Poster presentation

デキストラン結合タンパク質のアフィニティータグとしての利用
高原出帆; 田上貴祥; 奥山正幸
令和6年度日本応用糖質科学会北海道支部講演会, 03 Feb. 2025, Japanese, Poster presentation

Rhodococcus erythropolis NBRC100887株由来GH71酵素の構造解析と基質探索
小川真央; 田上貴祥; 奥山正幸
令和6年度日本応用糖質科学会北海道支部講演会, 03 Feb. 2025, Japanese, Poster presentation

Kribbella flavida由来GH15酵素 Kfla3969の発現系の構築および機能解析
眞鍋佑輔; 田上貴祥; 奥山正幸
日本応用糖質科学会2024年度大会, 26 Sep. 2024, Japanese
25 Sep. 2024 - 27 Sep. 2024

植物GH1 β-グルコシダーゼのラミナリオリゴ糖鎖長特異性の分子機構
宇野裕弥; 佐分利亘; 田上貴祥; 森春英
日本応用糖質科学会2024年度大会, 26 Sep. 2024, Japanese, Oral presentation
25 Sep. 2024 - 27 Sep. 2024

ブタの血清に含まれるα-グルコシダーゼのcDNAクローニングと機能構造解析
田上貴祥; 渡邊 憲; 枇杷千尋; 奥山正幸
第61回 日本生化学会北海道支部例会 日本生化学北海道支部・日本生物物理学会北海道支部 合同シンポジウム, 20 Jul. 2024, Japanese, Nominated symposium
[Invited]

L-glucoside 代謝関連酵素の探索，機能構造解析
奥山正幸; 田上貴祥
第1回 L-Glucose研究会, 13 Apr. 2024, Japanese, Oral presentation

An active site of osteopontin restoring the endometrial epidermal growth factor profile and fertility in repeat breeder dairy cows.
T. Tanida; T. Tagami; Y. Yanagawa; S. Katagiri
International Embryo Technology Science 50th Annual Conference, English, Oral presentation
09 Jan. 2024 - 12 Jan. 2024

デンプン代謝関連酵素群の機能と構造の多様性
田上貴祥
2023年度日本農芸化学会北海道支部第２回学術講演会, 09 Dec. 2023, Japanese, Invited oral presentation
[Invited]

α-グルコシダーゼの一般酸塩基触媒残基変異酵素を用いたグルコシルエステルの合成
藤井大河; 田上貴祥; 木村淳夫; 奥山正幸
令和4年度日本農芸化学会 北海道・東北支部 合同支部会, 21 Sep. 2022, Japanese, Oral presentation

ブタ血清 maltase-glucoamylase の保存方法および粗精製方法の検討
枇杷千尋; 田上貴祥; 奥山正幸
令和4年度日本農芸化学会 北海道・東北支部 合同支部会, 21 Sep. 2022, Japanese, Oral presentation

デキストラン代謝遺伝子群にコードされた solute-binding protein の機能および結晶構造の解析
宮園駿介; 田上貴祥; 奥山正幸
令和4年度日本農芸化学会 北海道・東北支部 合同支部会, 21 Sep. 2022, Japanese, Oral presentation

GH55 β-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
日本応用糖質科学会2022年度大会, 01 Sep. 2022, Japanese, Oral presentation
ポスター賞受賞

Kribbella flavida由来GH31 α-1,3-イソマルトシダーゼの基質特異性と結晶構造の解析
楊恵佳; 田上貴祥; 木村淳夫; 奥山正幸
日本応用糖質科学会2022年度大会, 01 Sep. 2022, Japanese, Oral presentation

Bifidobacterium adolescentis由来α-ガラクトシダーゼの基質特異性
大江 剛平; 奥山 正幸; 田上 貴祥; 中川 雄登; 菊池 麻子; 木村 淳夫
令和3年度日本応用糖質科学会北海道支部講演会, 27 Jan. 2022, Japanese, Poster presentation

デキストラン代謝関連遺伝子群にコードされたsolute-binding proteinの特異性解析
宮園駿介; 田上貴祥; 奥山正幸; 木村淳夫
令和3年度日本応用糖質科学会北海道支部講演会, 27 Jan. 2022, Japanese, Poster presentation

多糖資化遺伝子群に含まれる2種のSolute-binding proteinの機能構造解析
宮園駿介; 田上貴祥; 奥山正幸; 木村淳夫
日本応用糖質科学会令和3年度大会, Sep. 2021, Japanese, Oral presentation

Bifidobacterium adolescentis由来α−ガラクトシダーゼの機能解析
大江剛平; 奥山正幸; 田上貴祥; 木村淳夫
日本応用糖質科学会令和3年度大会, Sep. 2021, Japanese, Oral presentation

新奇α-L-glucosidaseの機能および結晶構造の解析
猪内 里花子; 田上 貴祥; 奥山 正幸; 木村 淳夫
日本応用糖質科学会令和3年度大会, Sep. 2021, Japanese, Oral presentation

Isomaltose glucohydrolase に特徴的なPhe290の機能解析
田上 貴祥; 古永 雄太; 奥山 正幸; 木村 淳夫
日本農芸化学会2021年度大会, Mar. 2021, Japanese, Oral presentation

α-グルコシダーゼによるα-マンノシル基転移反応の受容体特異性の解析
藤井大河; 床波 篤; 田上貴祥; 奥山正幸; 木村淳夫
令和2年度日本応用糖質科学会北海道支部会, Jan. 2021, Japanese, Poster presentation

糖質加水分解酵素ファミリー97 α-ガラクトシダーゼの糖転移反応の解析
岡本菜央; 菊池麻子; 田上貴祥; 奥山正幸; 木村淳夫
令和2年度日本応用糖質科学会北海道支部会, Jan. 2021, Japanese, Poster presentation

タマネギのフルクトオリゴ糖分解に関わる酵素の性質
乕田春佳; 上野敬司; 髙木惇生; 奥山正幸; 田上貴祥; 木村淳夫; 義平大樹; 小野寺秀一
令和2年度日本応用糖質科学会北海道支部会, Jan. 2021, Japanese, Poster presentation

糸状菌 α-グルコシダーゼ B への部位特異的飽和変異導入が生成物特異性に与える影響
床波 篤; Min Ma; 奥山 正幸; 田上 貴祥; 木村 淳夫
2020年度日本農芸化学会北海道支部/第50回日本栄養・食糧学会北海道支部合同学術講演会, Dec. 2020, Japanese, Oral presentation

組換え大腸菌を用いたラフィノースの細胞工学的生産
松井亨太; 奥山正幸; 田上貴祥; 木村淳夫
2020年度日本農芸化学会北海道支部/第50回日本栄養・食糧学会北海道支部合同学術講演会, Dec. 2020, Japanese, Oral presentation

GH13 に発見された α-1,3-グルコシダーゼの基質特異性を支配する構造因子
中山 英里香; 田上 貴祥; 奥山 正幸; 木村 淳夫
2020年度日本農芸化学会北海道支部/第50回日本栄養・食糧学会北海道支部合同学術講演会, Dec. 2020, Japanese, Oral presentation

組換えオステオポンチンによるリピートブリーダー牛の子宮内膜上皮成長因子（EGF）濃度正常化と受胎性回復効果の検証
佐藤 弘子; Hay Mar Kyaw; 栁川 洋二郎; 永野 昌志; 田上 貴祥; 片桐 成二
第61回日本卵子学会, Oct. 2020, Japanese, Oral presentation

Capacity of milk osteopontin to normalize endometrial epidermal growth factor profile in repeat breeder dairy cows
Hay Mar Kyaw; 栁川洋二郎; 田上貴祥; 永野昌志; 片桐成二
第163回 日本獣医学会学術集会, Sep. 2020, English, Oral presentation

New treatment option to improve fertility by intravaginal infusion of recombinant osteopontin in repeat breeder dairy cows
佐藤弘子; 田上貴祥; 栁川洋二郎; 永野昌志; 片桐成二
8th Sapporo Summer Symposium for One Health, Sep. 2020, English, Poster presentation

Effect of milk osteopontin on the normalization of endometrial epidermal growth factor profile in repeat breeder dairy cows
Hay Mar Kyaw; Takayoshi Tagami; Yojiro Yanagawa; Masashi Nagano; Seiji Katagiri
8th Sapporo Summer Symposium for One Health, Sep. 2020, English, Poster presentation

細菌 trehalose 6-phosphate hydrolaseの活性中心に結合するリン酸イオンおよび基質との複合体構造の分子解析
猪内 里花子; Klahan Patcharapa; 田上 貴祥; 田口陽大; 佐分利 亘; 森 春英; 奥山 正幸; 木村 淳夫
日本応用糖質科学会令和2年度大会, Sep. 2020, Japanese, Poster presentation

GH97 α-グルコシドヒドロラーゼが示す多様な基質特異性: 新奇な基質認識を与える2酵素と分子機構
菊池 麻子; 田上 貴祥; 奥山 正幸; 木村 淳夫
日本応用糖質科学会令和2年度大会, Sep. 2020, Japanese, Poster presentation

α-グルコシダーゼを用いた α-マンノシルオリゴ糖の合成
前山和輝; 田上貴祥; 奥山正幸; 木村淳夫
日本農芸化学会2020年度大会, Mar. 2020, Japanese, Others

ブタ血清由来マルターゼ-グルコアミラーゼを構成する2 つのサブユニットは独立に機能する
渡邊 憲; 田上貴祥; 奥山正幸; 木村淳夫
日本農芸化学会2020年度大会, Mar. 2020, Japanese, Others

ニゲロース特異的な Solute-binding protein の発見
田上貴祥; 壷井芙美; 佐藤宏樹; 奥山正幸; 佐分利亘; 森 春英; 木村淳夫
日本農芸化学会2020年度大会, Mar. 2020, Japanese, Others

放線菌由来 GH13_30 α-グルコシダーゼの機能解析
佐分利亘; 田上貴祥; 壷井芙美; 奥山正幸; 木村淳夫; 森 春英
日本農芸化学会2020年度大会, Mar. 2020, Japanese, Others

治療試験用オステオポンチンの調製と野外試験の現状
田上 貴祥
第112回 日本繁殖生物学会, Sep. 2019, Japanese, Nominated symposium
[Invited], [Domestic Conference]

デキストランデキストリナーゼのSer923の機能解析
嶌村有季乃; 田上貴祥; 奥山正幸; 木村淳夫
日本応用糖質科学会令和元年度大会, Sep. 2019, Japanese, Oral presentation
[Domestic Conference]

β-Fructofuranosidaseの糖転移作用：飽和変異導入による改変
髙木惇生; 奥山正幸; 田上貴祥; 木村淳夫
日本応用糖質科学会令和元年度大会, Sep. 2019, Japanese, Oral presentation
[Domestic Conference]

組み換えオステオポンチンによる牛子宮内膜上皮成長因子濃度の正常化効果の検証
佐藤 弘子; Hay Mar KYAW; 栁川 洋二郎; 永野 昌志; 田上 貴祥; 片桐 成二
第112回 日本繁殖生物学会, Sep. 2019, Japanese, Oral presentation
[Domestic Conference]

Bacteroides thetaiotaomicron由来GH97酵素：5つの機能未知パラログの解析
菊池麻子; 中川雄登; 奥山正幸; 田上貴祥; 木村淳夫
日本農芸化学会2019年度大会, Mar. 2019, Japanese, Oral presentation
[Domestic Conference]

Bacteroides thetaiotaomicron由来α-galactosidase(BtGal97a)の天然基質に関する研究
中川雄登; 松永夏奈; 菊池麻子; 奥山正幸; 田上貴祥; 木村淳夫
日本農芸化学会2019年度大会, Mar. 2019, Japanese, Oral presentation
[Domestic Conference]

ブタ血清由来マルターゼ-グルコアミラーゼを形成する酵素ユニットの機能
渡邊 憲; 田上貴祥; 奥山正幸; 木村淳夫
日本農芸化学会2019年度大会, Mar. 2019, Japanese, Oral presentation
[Domestic Conference]

ブタ血清由来α-グルコシダーゼの cDNA クローニングと組換え酵素の機能解析
渡邊憲; 田上貴祥; 奥山正幸; 木村淳夫
公益社団法人 日本農芸化学会 東北・北海道合同支部大会 (東北支部第 153 回大会), 23 Sep. 2018, Japanese, Oral presentation
[Domestic Conference]

改変 β-fructofuranosidase による 6-ケストースの合成および精製
髙木惇生; 芹沢領; 奥山正幸; 田上貴祥; 上野敬司; 小野寺秀一; 木村淳夫
公益社団法人 日本農芸化学会 東北・北海道合同支部大会 (東北支部第 153 回大会), 23 Sep. 2018, Japanese, Oral presentation
[Domestic Conference]

Structural and functional investigation of starch-metabolizing enzymes
Takayoshi Tagami
13th International Symposium of the Protein Society of Thailand, 07 Aug. 2018, English, Invited oral presentation
[Invited], [International presentation]

α-galactosidaseに想定される新奇な基質認識機構
菊池 麻子; 田上 貴祥; 奥山 正幸; 木村 淳夫
日本農芸化学会2018年度大会, 2018, Japanese

デキストラン デキストリナーゼの多糖合成に関与するアミノ酸の同定
佐々木 優希; 熊谷 裕也; 貞廣 樹里; ラング ビーラヌッチ; 田上 貴祥; 奥山 正幸; 木村 淳夫
日本農芸化学会2018年度大会, 2018, Japanese

Amino acid residues to govern substrate recognition of trehalose-6-phosphate hydrolase from Streptococcus mutans NBRC13955
Patcharapa KLAHAN; Masayuki OKUYAMA; Takayoshi TAGAMI; Atsuo KIMURA
日本農芸化学会2018年度大会, 2018, Japanese

Enzymatic characterization of trehalose-6-phosphate hydrolase from Streptococcus mutants NBRC13955
P. Klahan; M. Okuyama; T. Tagami; A. Kimura
12th Carbohydrate Bioengineering Meeting, 23 Apr. 2017, English, Poster presentation
[International presentation]

a-Glucosidase newly found in Aspergillus niger displays high specificity to a-(1→3)-glucosidic linkage
M. Ma; M. Okuyama; T. Tagami; A. Kimura
12th Carbohydrate Bioengineering Meeting, 23 Apr. 2017, English, Poster presentation
[International presentation]

Characterization of thermostable α-glucosidase from Geobacillus caldoxylosilyticus NBRC 107762, which displays remarkable transglucosylation
Min MA; Masayuki OKUYAMA; Takayoshi TAGAMI; Atsuo KIMURA
日本農芸化学会2017年度大会, 2017, Japanese

シングルアンカー型イソマルトメガロ糖におけるケルセチン-3-O-β-グルコシド可溶化に寄与する糖質構造の決定
土生 慎二; ラング ビーラヌッチ; 熊谷 裕也; 貞廣 樹里; 植草 聡太; 田上 貴祥; 奥山 正幸; 原 博; 木村 淳夫
日本農芸化学会2017年度大会, 2017, Japanese

Vibrio由来glycoside hydrolase family 17の機能未知なC末端領域は酵素活性の発現に必須である
熊谷 祐也; 田上 貴祥; 奥山 正幸; 木村 淳夫
日本農芸化学会2017年度大会, 2017, Japanese

新奇な基質認識機構を有するα-galactosidaseに関する研究
菊池麻子; 奥山正幸; 田上貴祥; 木村淳夫
平成２９年度 日本農芸化学会 北海道支部第１回講演会, 2017, Japanese

GH15 isomaltose glucohydrolaseの基質認識に関わる構造因子の同定
田上貴祥; 陳明皓; 奥山正幸; 岩_智仁; 田中良和; 姚閔; 木村淳夫
日本応用糖質科学会平成29年度大会, 2017, Japanese

Structure element to regulate transglucosidation specificity of Aspergillus niger α-glucosidase
M. Ma; M. Okuyama; M. Sato; T. Tagami; H. Mori; A. Kimura
6th Symposium on the Alpha-Amylase Family, 11 Sep. 2016, English, Poster presentation
[International presentation]

Reaction mechanism of glycosidases by studying 1,5-anhydro-d-fructose (AF) production
J. Maneesan; H. Matsuura; T. Tagami; H. Mori; A. Kimura
28th International carbohydrate symposium, 17 Jul. 2016, English, Poster presentation
[International presentation]

Purification and characterization of a-1,3-glucosidase from Aspergillus niger
M. Ma; T. Tagami; M. Okuyama; A. Kimura
28th International carbohydrate symposium, 17 Jul. 2016, English, Poster presentation
[International presentation]

環状四糖[cyclo-(1→6)-(α-nigerosyl-nigerosyl)]の分解を担う2種の糖質加水分解酵素の発見
田上貴祥; 貞廣樹里; 奥山正幸; 木村淳夫; 岩崎智仁
日本農芸化学会2016年度大会, 2016, Japanese

Kribbella flavida由来isomaltosyltransferaseの機能解析
宮野江梨; 田上貴祥; 奥山正幸; 木村淳夫
日本農芸化学会2016年度大会, 2016, Japanese

Efficient production and characterization of α-1,3-glucosidase from Aspergillus niger
Min MA; Takayoshi TAGAMI; Masayuki OKUYAMA; Atsuo KIMURA
日本農芸化学会2016年度大会, 2016, Japanese

ブタロース肉とヒレ肉におけるトリポリリン酸加水分解活性の比較
岩崎智仁; 大塚創太; 田上貴祥; 舩津保浩; 石下真人
日本畜産学会第121回大会, 2016, Japanese

GH97 α-galactosidase求核触媒変異酵素が触媒する糖転移反応の反応機構
松永 夏奈; 奥山 正幸; 渡辺 健一; 田上 貴祥; 山下 恵太郎; 姚 閔; 森 春英; 木村 淳夫
日本応用糖質科学会平成28年度大会, 2016, Japanese

長鎖阻害剤の利用による植物a-グルコシダーゼの機能構造相関の解明
田上貴祥; 山下恵太郎; 奥山正幸; 森春英; 姚 閔; 木村淳夫
平成27年度 応用糖質科学シンポジウム, 18 Sep. 2015, Japanese, Nominated symposium
[Invited], [Domestic Conference]

Structural and biochemical studies of sugar beet a-glucosidase exhibiting high specificity for long-chain substrates
T. Tagami; K. Yamashita; M. Okuyama; H. Mori; M. Yao; A. Kimura
11th Carbohydrate Bioengineering Meeting, 10 May 2015, English, Poster presentation
[International presentation]

A transglycosylation of catalytic nucleophile mutant of GH97 a-galactosidase with an external nucleophile
M. Okuyama; K. Matsunaga; K. Watanabe; T. Tagami; K. Yamashita; H. Mori; M. Yao; A. Kimura
11th Carbohydrate Bioengineering Meeting, 10 May 2015, English, Poster presentation
[International presentation]

テンサイa-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; 姚 閔; 木村淳夫
日本農芸化学会2015年度大会, 2015, Japanese

食肉タンパク質の溶解性と熱ゲル化特性に及ぼすアミノ酸添加の影響
船津保浩; 岩_智仁; 田上貴祥
FOOMA JAPAN 2015, 2015, Japanese

Production of 1,5-anhydro-d-fructose by a-glucosidase belong to glycoside hydrolase family 13 and 31
Janjira Maneesan; Hideyuki Matsuura; Takayoshi Tagami; Momoko Kobayashi; Masayuki Okuyama; Haruhide Mori; Atsuo Kimura
日本農芸化学会2014年度大会, 2014, Japanese

筋損傷部位における酸化タンパク質領域の可視化
岩_智仁; 石崎恵梨; 田上貴祥; 美名口順; 竹花一成; Miranda Grounds; Peter Arthur
日本畜産学会第118回大会, 2014, Japanese

放線菌Kribbella flavidaによるオリゴ糖合成の検討
田上貴祥; 奥山正幸; 岩_智仁; 木村淳夫
日本農芸化学会平成26年度北海道支部・東北支部合同支部会, 2014, Japanese

Enzymatic synthesis of acarviosyl-maltooligosaccharides and their inhibitory effects on a-glucosidase
T. Tagami; Y. Tanaka; H. Mori; M. Okuyama; A. Kimura
26th International Carbohydrate Symposium, 22 Jul. 2012, English, Oral presentation
[International presentation]

Structural insights into specificity for long-chain substrates of sugar beet a-glucosidase
T. Tagami; K. Yamashita; M. Okuyama; H. Mori; M. Yao; A. Kimura
Plant and Seaweed Polysaccharides Symposium 2012, 17 Jul. 2012, English, Oral presentation
[International presentation]

Acarviosyl-maltooligosaccharideの酵素合成とテンサイa-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
日本応用糖質科学会平成24年度大会, 2012, Japanese

Aspergillus niger由来a-glucosidaseの基質特異性に関与するアミノ酸残基の解析
佐藤恵美; 田上貴祥; 奥山正幸; 森春英; 木村淳夫
日本農芸化学会北海道支部平成24年度支部講演会, 2012, Japanese

Structural element determining the diverse specificities for chain-length of substrate in glycoside hydrolase family 31 a-glucosidases
T. Tagami; M. Okuyama; H. Mori; A. Kimura
9th Carbohydrate Bioengineering Meeting, 15 May 2011, English, Poster presentation
[International presentation]

GH 31 a-glucosidase における基質の鎖長認識機構の解明
田上貴祥; 奥山正幸; 森 春英; 木村淳夫
日本農芸化学会北海道支部平成23年度夏期シンポジウム, 2011, Japanese

GH family 31に属するAspergillus niger 由来a-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
日本応用糖質科学会平成23年度大会, 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; 木村敦夫
日本応用糖質科学会平成23年度大会, 2011, Japanese

Shifting specificity of Aspergillus niger a-glucosidase from short-chain substrates to long-chain substrates
T. Tagami; T. Nishimura; M. Okuyama; H. Mori; A. Kimura
Plant Polysaccharide and Applied Glycoscience Workshop 2010, 29 Jul. 2010, English, Oral presentation
[International presentation]

GH family 31 a-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2010年度大会, 2010, Japanese

Aspergillus niger a-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
日本応用糖質科学会平成22年度大会, 2010, Japanese

Specificity element recognizing long-chain substrates in GH family 31 a-glucosidase derived from sugar beet (Beta vulgaris L.)
T. Tagami; M. Okuyama; H. Nakai; H. Mori; K. Taguchi; A. Kimura
8th Carbohydrate Bioengineering Meeting, 10 May 2009, English, Poster presentation
[International presentation]

テンサイa-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
日本応用糖質科学会平成21年度大会, 2009, Japanese

テンサイa-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
日本応用糖質科学会平成20年度大会, 2008, Japanese

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

バイオテクノロジー学特論, 2024年, 修士課程, 農学院

バイオテクノロジー学特論演習, 2024年, 修士課程, 農学院

応用分子生物学特論, 2024年, 修士課程, 農学院

応用分子生物学特論演習, 2024年, 修士課程, 農学院

生物学Ⅰ, 2024年, 学士課程, 全学教育

応用生命科学概論, 2024年, 学士課程, 農学部

蛋白質工学, 2024年, 学士課程, 農学部

応用生命科学演習Ⅰ, 2024年, 学士課程, 農学部

化学概論, 2024年, 学士課程, 農学部

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

Apr. 2021
日本蛋白質科学会

牛子宮での上皮成長因子異常におけるToll様受容体4活性化の意義と治療への応用
科学研究費助成事業
01 Apr. 2023 - 31 Mar. 2028
片桐 成二; 柳川 洋二郎; 杉浦 智親; 奥山 みなみ; 平山 博樹; 永野 昌志; 田上 貴祥
日本学術振興会, 基盤研究(A), 北海道大学, 23H00353

細胞質雄性不稔性発現における翻訳後メカニズムの研究
科学研究費助成事業
01 Apr. 2024 - 31 Mar. 2027
久保 友彦; 北崎 一義; 田上 貴祥; 松平 洋明
日本学術振興会, 基盤研究(B), 北海道大学, 24K01726

Characterization of carbohydrate-binding proteins and their applications
Grants-in-Aid for Scientific Research
01 Apr. 2023 - 31 Mar. 2026
田上 貴祥
本研究は、これまで積極的な研究がなされてこなかった糖質輸送タンパク質に焦点をあてたものである。なかでも原核生物において糖輸送の初発段階をになう溶質結合タンパク質(Solute-binding protein, SBP)の特異性と構造を網羅的に理解し、応用につなげることを目的としている。
令和5年度は、新規SBPの探索を実施した。原核生物のゲノム情報を参照し、周辺遺伝子情報からリガンド候補を推定できた3種のSBP（SBP1, SBP2, SBP3と略）を選択した。SBP1については、大腸菌による組換えタンパク質の生産と精製および等温滴定カロリメトリーを用いた相互作用解析を実施し、SBP1がニゲロオリゴ糖に特異性を有することを明らかにした。SBP1と既知のニゲロオリゴ糖結合SBPとのアミノ酸配列類似性は67%であり、4%のギャップを含んでいた。SBP1の結晶化条件探索もすでに開始している。今後は既知SBPとの立体構造比較を通じて、両者のニゲロオリゴ糖認識の分子機構を明らかにする計画である。SBP2については、大腸菌による組換えタンパク質の生産系を確立した。SBP3については、遺伝子を取得して発現用プラスミドベクターを構築済みであり、大腸菌による組換えタンパク質生産実験に移行中である。
また令和5年度は、IG2-SBPのアフィニティー精製タグとしての応用を検証した。IG2-SBP融合タンパク質は市販の安価なクロマトグラフィー担体に特異的に吸着し、グルコースによって解離することを明らかにした。現在のアフィニティー精製タグの主流であるヒスチジンタグよりも、簡便かつ高倍率で目的タンパク質を精製できることを明らかにした。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 23K05034

牛の精漿蛋白質による子宮機能調節機序の解明と受胎性改善技術への応用
科学研究費助成事業 基盤研究（A）
Apr. 2019 - Mar. 2024
片桐 成二
日本学術振興会, Competitive research funding

Problem-based researches utilized by novel megalosaccharides dissolving poorly-soluble BCS II compounds
Grants-in-Aid for Scientific Research Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
07 Oct. 2019 - 31 Mar. 2023
木村 淳夫; 橋床 泰之; 崎浜 靖子; 奥山 正幸; 田上 貴祥
我々は世界で初めてメガロ糖（MS）の生産に成功した。性質を調べると、BCS IIに属す化合物（難水溶性・高膜透過性の薬剤や食品素材など）を可溶化する画期的な機能が発見された。またMSは「BCS II化合物を溶質とする糖質水溶化剤」と捉えることもできた。一方、難溶性ベンジル系アゾ色素もBCS IIに属し、かつ「東南アジア諸国における名高い環境汚染物質」である点に注目し、MSとアゾ分解酵素を組合せることで、実験室レベルではあるが、色素の可溶化と酵素分解に成功した。以上は初めて生産したMS、すなわち従来型MSの知見である。極最近に従来型MSより高機能な新奇MS（新型MS）を発見した。本申請では、新型MSによるアゾ色素の酵素分解を目的とし、現地試験をタイで実施する。最終的な到達目標はアゾ色素の汚染解消（すなわち環境問題解決への貢献）である。
本年度は、新型コロナウイルス感染症の流行および相手国の政情不安から渡航が困難となり、現地調査に大きな支障が生じ、計画の遂行に予想外の大きな遅れが発生した。本状況下で得られた成果を述べる。多糖選抜および新型MS調製の項目に関し、昨年度に行った研究計画上の工夫（調査が不可な期間は、日タイ共通植物種を対象に研究を先行。渡タイ可能時に現地で結果検証）により研究を進めた。共通植物種から取得したMSが示すBCS II化合物の可溶化能を測定し、優れた能力を有する新奇MSを見出した。さらに調査対象の植物種を増やし、多糖調製・MS分離・構造と機能の解析を継続している。なお、現地で実施が必要なMS高機能化および色素汚染解消に関する計画に大幅な遅延が生じているが、渡航開始後に打開を目指したい。
Japan Society for the Promotion of Science, Fund for the Promotion of Joint International Research (Fostering Joint International Research (B)), Hokkaido University, 19KK0147

乳牛の低受胎対策技術の実行可能性検証事業
日本中央競馬会畜産振興事業
Apr. 2021 - Mar. 2023
公益財団法人全国競馬・畜産振興会, Coinvestigator

O-マンノース型糖鎖の酵素合成と分解酵素探索
科学研究費助成事業 若手研究
Apr. 2019 - Mar. 2022
田上 貴祥
日本学術振興会, Principal investigator, Competitive research funding

Further development of megalosaccharide research: synthesis and application of novel megalosaccharides displaying excellent functions
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
01 Apr. 2017 - 31 Mar. 2021
KIMURA Atsuo
We succeeded in production of megalosaccharides and found their valuable function to solubilize water-insoluble compounds. However, the period of our research is very short, so that we have many problems that must be solved. This project challenges a resolution of three important problems, from which we will obtain the fundamental knowledge about megalosaccharides. Furthermore, it also contributes to the development of application research on megalosaccharides. The purposes of this program are 1) analysis of molecular mechanism of polysaccharide-forming enzyme to produce megalosaccharide, 2) synthesis of new megalosaccharide with high functionality, and 3) improvement of azo-dye pollution using megalosaccharide.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 17H03801

タンパク質工学による新奇オリゴ糖合成酵素の作出
研究助成
Oct. 2018 - Mar. 2020
田上 貴祥
公益財団法人栗林育英学術財団, Principal investigator, Competitive research funding

乳牛の低受胎対策新規技術開発事業
日本中央競馬会畜産振興事業
Apr. 2017 - Mar. 2020
片桐 成二
日本中央競馬会, Competitive research funding

放線菌由来α-1,3-グルカナーゼの構造機能相関の解明
研究奨励金
Dec. 2017 - Mar. 2019
田上 貴祥
公益財団法人ホクト生物科学振興財団, Principal investigator, Competitive research funding

哺乳動物の血液・精液に存在する糖質分解酵素の機能解明
科学研究費助成事業 若手研究(B)
Apr. 2017 - Mar. 2019
田上 貴祥
日本学術振興会, Principal investigator, Competitive research funding

O-マンノース型糖鎖の新規合成法の確立
研究助成金
Apr. 2014 - Mar. 2016
田上 貴祥
公益財団法人酪農学園後援会, Principal investigator, Competitive research funding

食肉加工特性の可視化技術の開発
研究奨励金
Apr. 2014 - Mar. 2015
田上 貴祥
公益財団法人酪農育英会, Principal investigator, Competitive research funding

レボグルコサンを経由する新奇デンプン代謝経路の解明
研究助成（奨励）
Apr. 2014 - Mar. 2015
田上 貴祥
公益財団法人秋山記念生命科学振興財団, Principal investigator, Competitive research funding

放線菌Kribbella flavidaによる糖合成系の開発
フードイノベーション創造支援事業 研究シーズ発掘助成金
Aug. 2013 - Feb. 2014
田上 貴祥
公益財団法人北海道科学技術総合振興センター, Principal investigator, Competitive research funding

第 65 回澱粉研究懇談会
06 Jun. 2025
Lecturer
澱粉研究懇談会 世話人会

あぐり大学第41回講座「わたしたちのくらしと酵（こう）素」
07 Sep. 2024
Lecturer
北海道大学農学部、北海道新聞社

北海道札幌藻岩高等学校指定科目「環境教育講座」
06 Sep. 2019
Lecturer

第22期 ホイスコーレ札幌
05 Jun. 2019
Lecturer

北海道札幌藻岩高等学校指定科目「環境教育講座」
14 Sep. 2018
Lecturer

第29回三省堂サイエンスカフェin札幌〜日本農芸化学会 北海道シリーズ10〜
04 Aug. 2018
Planner

北海道札幌藻岩高等学校指定科目「環境教育講座」
15 Sep. 2017
Lecturer

えべつ市民カレッジ 平成27年度ふるさと江別塾
24 Oct. 2015
Lecturer

北海道札幌開成高等学校指定科目「先端科学特論」
29 Aug. 2015
Lecturer

北海道石狩南高等学校 「出前講義」
17 Jun. 2015
Lecturer

北海道札幌開成高等学校指定科目「先端科学特論」
31 Aug. 2014
Lecturer

北海道札幌開成高等学校指定科目「先端科学特論」
24 Aug. 2013
Lecturer