Mar. 2023, Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA), JSBBA Award　　　　　　　　　　　　　　　
Studies on biosynthetic enzymes leading structural and functional diversities to microbial natural products.

Mar. 2022, 日本農芸化学会, トピックス賞　　　　　　　　　　　　　　　
微細藻類由来DHA合成酵素の炭素鎖伸長反応の解析
仲間 陸;小林 飛悠;大塚 慎;佐藤 康治;小笠原 泰志;大利 徹

Mar. 2021, 日本農芸化学会, トピックス賞　　　　　　　　　　　　　　　
ポリグルタミン酸生合成におけるエピメリ化酵素の同定
加藤 陽菜多、小笠原 泰志、大利 徹

Mar. 2019, 日本農芸化学会, トピックス賞　　　　　　　　　　　　　　　
in vitro解析による多価不飽和脂肪酸生合成酵素の炭素鎖長制御機構の解明
林 祥平;小笠原 泰志;佐藤 康治;丸山 千登勢;濱野 吉十;氏原 哲朗;大利 徹

Mar. 2017, 日本農芸化学会, トピックス賞　　　　　　　　　　　　　　　
An unprecedented glutamate epimerase for bacterial peptidoglycan biosynthesis
Ruoyin FENG;Yasuharu Satoh;Yasushi Ogasawara;Tohru Yoshimura;Tohru Dairi

Jun. 2015, 日本生物工学会, 第23回生物工学論文賞　　　　　　　　　　　　　　　
New gene responsible for para-aminobenzoate biosynthesis
Y. Satoh;M. Kuratsu;D. Kobayashi;T. Dairi

2012, 酵素応用シンポジウム運営委員会, 酵素応用シンポジウム奨励賞　　　　　　　　　　　　　　　
大利 徹

2011, 長瀬財団, Nagase Foundation Award　　　　　　　　　　　　　　　
大利 徹

2010, 日本放線菌学会, 日本放線菌学会 学会賞　　　　　　　　　　　　　　　
大利 徹

2004, 住木・梅澤記念賞　　　　　　　　　　　　　　　
Japan

2004, Sumiki & Umezawa Award from Japan Antibiotic Research Association　　　　　　　　　　　　　　　

2002, 日本農芸化学会論文賞　　　　　　　　　　　　　　　
Japan

2000, 日本農芸化学会奨励賞　　　　　　　　　　　　　　　
Japan

2000, Encouragement Award from Japan Society for Bioscience, Biotechnology, and Agrochemistry　　　　　　　　　　　　　　　

1999, とやま賞　　　　　　　　　　　　　　　
Japan

1999, TOYAMA Award from Toyama Hitodukuri Foundation　　　　　　　　　　　　　　　

1998, 日本放線菌学会浜田賞　　　　　　　　　　　　　　　
Japan

1998, Encouragement Award from The Society for Actinomycetes Japan　　　　　　　　　　　　　　　

Biosynthesis of lactacystin as a proteasome inhibitor.
Takeshi Tsunoda, Shunkichi Furumura, Haruka Yamazaki, Chitose Maruyama, Yoshimitsu Hamano, Yasushi Ogasawara, Tohru Dairi
Communications chemistry, 8, 1, 9, 9, 13 Jan. 2025, [International Magazine]
English, Scientific journal, Lactacystin is an irreversible proteasome inhibitor isolated from Streptomyces lactacystinicus. Despite its importance for its biological activity, the biosynthesis of lactacystin remains unknown. In this study, we identified the lactacystin biosynthetic gene cluster by gene disruption and heterologous expression experiments. We also examined the functions of the genes encoding a PKS/NRPS hybrid protein (LctA), NRPS (LctB), ketosynthase-like cyclase (LctC), cytochrome P450 (LctD), MbtH-like protein (LctE), and formyltransferase (LctF) by in vivo and in vitro experiments. In particular, we demonstrated that LctF directly transferred the formyl group of 10-N-formyl tetrahydrofolate to CoA. The formyl group of formyl-CoA was then transferred to ACP1 by LctA_AT1 to form formyl-ACP1. This is the first example of an AT domain recognizing a formyl group. The formyl group is perhaps transferred to methylmalonate tethered on LctA_ACP2 to yield methylmalonyl-semialdehyde-ACP2. Then, it would be condensed with leucine bound to PCP in LctB by the C domain in LctA. Using a mimic compound, we confirmed that LctC catalyzed the formation of the cyclic α,α-disubstituted amino acid structure with concomitant release of the product from PCP. Thus, we figured out the overall biosynthesis of lactacystin including a novel role of a formyl group in a secondary metabolite.

Optimization of tyrosol-producing pathway with tyrosine decarboxylase and tyramine oxidase in high-tyrosine-producing Escherichia coli
Ning Shen, Yasuharu Satoh, Daisuke Koma, Hiroyuki Ohashi, Yasushi Ogasawara, Tohru Dairi
Journal of Bioscience and Bioengineering, 137, 2, 115, 123, Elsevier BV, Feb. 2024
Scientific journal

Identification of a new oligomycin derivative as a specific inhibitor of the alternative peptidoglycan biosynthetic pathway.
Shuhei Umetsu, Takeshi Tsunoda, Haruka Kiyanagi, Yuki Inahashi, Kenichi Nonaka, Tohru Dairi, Yasushi Ogasawara
The Journal of antibiotics, 10 Jan. 2024, [International Magazine]
English, Scientific journal, Peptidoglycan is an important macromolecule in bacterial cell walls to maintain cell integrity, and its biosynthetic pathway has been well studied. Recently, we demonstrated that some bacteria such as Xanthomonas oryzae, a pathogen causing bacterial blight of rice, used an alternative pathway for peptidoglycan biosynthesis. In this pathway, MurD2, a MurD homolog, catalyzed the attachment of L-Glu to UDP-MurNAc-L-Ala and MurL, which did not show homology to any known protein, catalyzed epimerization of the terminal L-Glu of the MurD2 product to generate UDP-MurNAc-L-Ala-D-Glu. Because the alternative pathway also operates in some other plant pathogens and opportunistic pathogens, specific inhibitors of the alternative pathway could function as pesticides and antibiotics for these pathogens. In this study, we searched for specific inhibitors of the alternative pathway from metabolites produced by actinomycetes and identified a new oligomycin-class polyketide, which was revealed to inhibit the MurD2 reaction, in culture broth of Micromonospora sp. K18-0097.

Peptide Epimerase Responsible for d-Amino Acid Introduction in Poly-γ-glutamic Acid Biosynthesis
Hinata Kato, Moeka Sakuta, Takeshi Tsunoda, Yu Nakashima, Hiroyuki Morita, Yasushi Ogasawara, Tohru Dairi
Biomacromolecules, 25, 1, 349, 354, American Chemical Society (ACS), 14 Dec. 2023
Scientific journal

Structure of lasso peptide epimerase MslH reveals metal-dependent acid/base catalytic mechanism
Yu Nakashima, Atsushi Kawakami, Yasushi Ogasawara, Masatoshi Maeki, Manabu Tokeshi, Tohru Dairi, Hiroyuki Morita
Nature Communications, 14, 1, Springer Science and Business Media LLC, 08 Aug. 2023
Scientific journal, Abstract

The lasso peptide MS-271 is a ribosomally synthesized and post-translationally modified peptide (RiPP) consisting of 21 amino acids with D-tryptophan at the C-terminus, and is derived from the precursor peptide MslA. MslH, encoded in the MS-271 biosynthetic gene cluster (msl), catalyzes the epimerization at the Cα center of the MslA C-terminal Trp21, leading to epi-MslA. The detailed catalytic process, including the catalytic site and cofactors, has remained enigmatic. Herein, based on X-ray crystallographic studies in association with MslA core peptide analogues, we show that MslH is a metallo-dependent peptide epimerase with a calcineurin-like fold. The crystal structure analysis, followed by site-directed mutagenesis, docking simulation, and ICP-MS studies demonstrate that MslH employs acid/base chemistry to facilitate the reversible epimerization of the C-terminal Trp21 of MslA, by utilizing two pairs of His/Asp catalytic residues that are electrostatically tethered to a six-coordination motif with a Ca(II) ion via water molecules.

Peptide epimerase-dehydratase complex responsible for biosynthesis of the linaridin class ribosomal peptides.
Wanlu Xiao, Takeshi Tsunoda, Chitose Maruyama, Yoshimitsu Hamano, Yasushi Ogasawara, Tohru Dairi
Bioscience, biotechnology, and biochemistry, 87, 11, 1316, 1322, 04 Aug. 2023, [International Magazine]
English, Scientific journal, Grisemycin, salinipeptin, and cypemycin belong to the linaridin class of ribosomally synthesized and posttranslationally modified peptides that contain multiple dehydrobutyrine and D-amino acid residues. The biosynthetic gene clusters of these linaridins lack obvious candidate genes for the dehydratase and epimerase required to introduce dehydrobutyrine and D-amino acid residues, respectively. However, we previously demonstrated that the grisemycin (grm) cluster contained cryptic dehydratase and epimerase genes by heterologous expression of this biosynthetic gene cluster in Streptomyces lividans and proposed that two genes (grmH and grmL) with unknown functions catalyze dehydration and epimerization reactions. In this study, we confirmed that both GrmH and GrmL, which were shown to constitute a protein complex by a co-purification experiment, were required to catalyze the dehydration, epimerization, and proteolytic cleavage of a precursor peptide GrmA by in vivo experiments. Furthermore, we demonstrated that GrmH/GrmL complex accepted salinipeptin and cypemycin precursor peptides, which possess three additional amino acids.

Studies on biosynthetic enzymes leading to structural and functional diversity of microbial natural products
Tohru Dairi
Bioscience, Biotechnology, and Biochemistry, 87, 8, 797, 808, Oxford University Press (OUP), 24 May 2023
Scientific journal, ABSTRACT

The primary metabolic pathways, for the most part, have been disclosed in Escherichia coli and Saccharomyces cerevisiae. These pathways were believed to be common among all microorganisms. However, after discovery of an alternative pathway for biosynthesis of isopentenyl diphosphate, the methylerythritol phosphate pathway, genome mining of alternative biosynthetic pathways for primary metabolites has been performed. My collaborators and I focused on the biosynthetic pathways of menaquinone and peptidoglycan because some microorganisms lack ortholog genes of the known biosynthetic pathways for these compounds. I also studied biosynthetic enzymes for secondary metabolites produced by actinomycetes and fungi because they include many unique enzymes. In this review, outlines of these studies are described.

N-Formimidoylation/-iminoacetylation modification in aminoglycosides requires FAD-dependent and ligand-protein NOS bridge dual chemistry.
Yung-Lin Wang, Chin-Yuan Chang, Ning-Shian Hsu, I-Wen Lo, Kuan-Hung Lin, Chun-Liang Chen, Chi-Fon Chang, Zhe-Chong Wang, Yasushi Ogasawara, Tohru Dairi, Chitose Maruyama, Yoshimitsu Hamano, Tsung-Lin Li
Nature communications, 14, 1, 2528, 2528, 03 May 2023, [International Magazine]
English, Scientific journal, Oxidized cysteine residues are highly reactive and can form functional covalent conjugates, of which the allosteric redox switch formed by the lysine-cysteine NOS bridge is an example. Here, we report a noncanonical FAD-dependent enzyme Orf1 that adds a glycine-derived N-formimidoyl group to glycinothricin to form the antibiotic BD-12. X-ray crystallography was used to investigate this complex enzymatic process, which showed Orf1 has two substrate-binding sites that sit 13.5 Å apart unlike canonical FAD-dependent oxidoreductases. One site could accommodate glycine and the other glycinothricin or glycylthricin. Moreover, an intermediate-enzyme adduct with a NOS-covalent linkage was observed in the later site, where it acts as a two-scissile-bond linkage facilitating nucleophilic addition and cofactor-free decarboxylation. The chain length of nucleophilic acceptors vies with bond cleavage sites at either N-O or O-S accounting for N-formimidoylation or N-iminoacetylation. The resultant product is no longer sensitive to aminoglycoside-modifying enzymes, a strategy that antibiotic-producing species employ to counter drug resistance in competing species.

Ergothioneine production by Corynebacterium glutamicum harboring heterologous biosynthesis pathways.
Takashi Hirasawa, Yuki Shimoyamada, Yukio Tachikawa, Yasuharu Satoh, Yusuke Kawano, Tohru Dairi, Iwao Ohtsu
Journal of bioscience and bioengineering, 135, 1, 25, 33, Jan. 2023, [Domestic magazines]
English, Scientific journal, In this study, Corynebacterium glutamicum was engineered to produce ergothioneine, an amino acid derivative with high antioxidant activity. The ergothioneine biosynthesis genes, egtABCDE, from Mycolicibacterium smegmatis were introduced into wild-type and l-cysteine-producing strains of C. glutamicum to evaluate their ergothioneine production. In the l-cysteine-producing strain, ergothioneine production reached approximately 40 mg L-1 after 2 weeks, and the amount was higher than that in the wild-type strain. As C. glutamicum possesses an ortholog of M. smegmatis egtA, which encodes an enzyme responsible for γ-glutamyl-l-cysteine synthesis, the effect of introducing egtBCDE genes on ergothioneine production in the l-cysteine-producing strain was evaluated, revealing that a further increase to more than 70 mg L-1 was achieved. As EgtBs from Methylobacterium bacteria are reported to use l-cysteine as a sulfur donor in ergothioneine biosynthesis, egtB from Methylobacterium was expressed with M. smegmatis egtDE in the l-cysteine-producing strain. As a result, ergothioneine production was further improved to approximately 100 mg L-1. These results indicate that utilization of the l-cysteine-producing strain and introduction of heterologous biosynthesis pathways from M. smegmatis and Methylobacterium bacteria are effective for improved ergothioneine production by C. glutamicum.

First direct evidence for direct cell-membrane penetrations of polycationic homopoly(amino acid)s produced by bacteria
Yamato Takeuchi, Kazunori Ushimaru, Kohei Kaneda, Chitose Maruyama, Takashi Ito, Kazuya Yamanaka, Yasushi Ogasawara, Hajime Katano, Yasuo Kato, Tohru Dairi, Yoshimitsu Hamano
Communications Biology, 5, 1, Springer Science and Business Media LLC, 26 Oct. 2022
Scientific journal, Abstract

Bacteria produce polycationic homopoly(amino acid)s, which are characterized by isopeptide backbones. Although the biological significance of polycationic homopoly(amino acid)s remains unclear, increasing attention has recently been focused on their potential use to achieve cellular internalization. Here, for the first time, we provide direct evidence that two representative bacterial polycationic isopeptides, ε-poly-l-α-lysine (ε-PαL) and ε-oligo-l-β-lysine (ε-OβL), were internalized into mammalian cells by direct cell-membrane penetration and then diffused throughout the cytosol. In this study, we used clickable ε-PαL and ε-OβL derivatives carrying a C-terminal azide group, which were enzymatically produced and then conjugated with a fluorescent dye to analyze subcellular localization. Interestingly, fluorescent proteins conjugated with the clickable ε-PαL or ε-OβL were also internalized into cells and diffused throughout the cytosol. Notably, a Cre recombinase conjugate with ε-PαL entered cells and mediated the Cre/loxP recombination, and ε-PαL was found to deliver a full-length IgG antibody to the cytosol and nucleus.

Biosynthetic Gene Cluster of Linaridin Peptides Contains Epimerase Gene
Wanlu Xiao, Yasuharu Satoh, Yasushi Ogasawara, Tohru Dairi
ChemBioChem, 23, 12, Wiley, 20 Jun. 2022, [Peer-reviewed], [Corresponding author], [International Magazine]
Scientific journal

Identification of Cyclopropane Formation in the Biosyntheses of Hormaomycins and Belactosins: Sequential Nitration and Cyclopropanation by Metalloenzymes
Xiaojun Li, Ryo Shimaya, Tohru Dairi, Wei‐chen Chang, Yasushi Ogasawara
Angewandte Chemie International Edition, 61, 7, Wiley, 07 Feb. 2022, [Peer-reviewed], ［Internationally co-authored］, [International Magazine]
Scientific journal

Identification of pulvomycin as an inhibitor of the futalosine pathway
Yasushi Ogasawara, Shuhei Umetsu, Yuki Inahashi, Kenichi Nonaka, Tohru Dairi
The Journal of Antibiotics, 20 Aug. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Discovery of an alternative pathway of peptidoglycan biosynthesis: A new target for pathway specific inhibitors
Yasushi Ogasawara, Tohru Dairi
Journal of Industrial Microbiology and Biotechnology, Oxford University Press (OUP), 11 Jun. 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal,
Peptidoglycan in bacterial cell walls is a biopolymer consisting of sugars and amino acids and plays important role in maintaining cell integrity from the environment. Its biosynthesis is a major target for antibiotics and the genes and enzymes involved in the biosynthetic pathway have been well studied. However, we recently identified an alternative pathway in the early stage of peptidoglycan biosynthesis in Xanthomonas oryzae, a plant pathogen causing bacterial blight disease of rice. The distribution of the alternative pathway is limited to relatively few bacterial genera that contain many pathogenic species, including Xylella and Stenotrophomonas, besides Xanthomonas. Thus, the alternative pathway is an attractive target for the development of narrow-spectrum antibiotics specific to pathogens. In this minireview, we summarize the discovery of the alternative pathway and identification of its specific inhibitors.

Flavonoids from Woodfordia fruticosa as potential SmltD inhibitors in the alternative biosynthetic pathway of peptidoglycan
Yuan-E Lee, Takeshi Kodama, Nwet Nwet Win, Dae-Won Ki, Nhat Nam Hoang, Chin Piow Wong, Khine Zar Wynn Lae, Hla Ngwe, Tohru Dairi, Hiroyuki Morita
Bioorganic & Medicinal Chemistry Letters, 36, 127787, 127787, Elsevier BV, Mar. 2021, [Peer-reviewed], [Corresponding author], ［Internationally co-authored］, [International Magazine]
English, Scientific journal, SmltD is an ATP-dependent ligase that catalyzes the condensation of UDP-MurNAc-L-Ala and L-Glu to form UDP-MurNAc-L-Ala-L-Glu, in the newly discovered peptidoglycan biosynthesis pathway of a Gram-negative multiple-drug-resistant pathogen, Stenotrophomonas maltophilia. Phytochemical investigation of the 70% ethanol extract from Woodfordia fruticosa flowers collected in Myanmar led to the identification of anti-SmltD active flavonoids, kaempferol 3-O-(6''-galloyl)-β-D-glucopyranoside (1), astragalin (2), and juglalin (3). Among them, 1 showed the most potent SmltD inhibitory activity. An enzyme steady-state kinetic study revealed that 1 exerted competitive inhibition with respect to ATP. The results of this study provided an attractive foundation for the further development of novel inhibitors of SmltD.

Identification of the peptide epimerase MslH responsible for d-amino acid introduction at the C-terminus of ribosomal peptides
Zhi Feng, Yasushi Ogasawara, Tohru Dairi
Chemical Science, 12, 7, 2567, 2574, Royal Society of Chemistry (RSC), 2021, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal,

The biosynthesis of d-tryptophan containing lasso peptide MS-271 involves the epimerization of a ribosomal peptide MslA catalyzed by a novel class of metal- and cofactor-independent peptide epimerase MslH.

High Production of Ergothioneine in Escherichia coli using the Sulfoxide Synthase from Methylobacterium strains
Tomoyuki Kamide, Shun Takusagawa, Naoyuki Tanaka, Yasushi Ogasawara, Yusuke Kawano, Iwao Ohtsu, Yasuharu Satoh, Tohru Dairi
Journal of Agricultural and Food Chemistry, 68, 23, 6390, 6394, American Chemical Society (ACS), 10 Jun. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, We previously constructed a heterologous production system for ergothioneine (ERG) in Escherichia coli using five ERG biosynthesis genes (egtABCDE) from Mycobacterium smegmatis. However, significant amounts of hercynine (HER), an intermediate of ERG, as ERG were accumulated, suggesting that the reaction of EgtB catalyzing the attachment of γ-glutamylcysteine (γGC) to HER to yield hercynyl-γ-glutamylcysteine sulfoxide was a bottleneck. In this study, we searched for other EgtBs and found many egtB orthologs in diverse microorganisms. Among these, Methylobacterium strains possessed EgtBs that catalyze the direct conversion of HER into hercynylcysteine sulfoxide with l-cysteine (l-Cys) as a sulfur donor, in a manner similar to those of acidobacterial CthEgtB and fungal Egt1. An in vitro study with recombinant EgtBs from Methylobacterium brachiatum and Methylobacterium pseudosasicola clearly showed that both enzymes accepted l-Cys but not γGC. We reconstituted the ERG production system in E. coli with egtB from M. pseudosasicola; ERG productivity reached 657 mg L-1.

Recent advances in functional analysis of polyunsaturated fatty acid synthases.
Shohei Hayashi, Yasuharu Satoh, Yasushi Ogasawara, Tohru Dairi
Current opinion in chemical biology, 59, 30, 36, 19 May 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid, eicosapentaenoic acid, and arachidonic acid are essential fatty acids for humans. PUFAs are biosynthesized by either desaturases/elongases from oleic acid or PUFA synthases from acetyl units. PUFA synthases are composed of three or four subunits, and each creates a specific PUFA even though the multiple catalytic domains in each subunit are very similar. We recently dissected these PUFA synthases by in vivo and in vitro experiments and elucidated how the enzymes control PUFA profiles. Moreover, for the first time, we converted a practical microalgal docosahexaenoic acid synthase into an eicosapentaenoic acid synthase based on the results.

Off-Loading Mechanism of Products in Polyunsaturated Fatty Acid Synthases.
Shohei Hayashi, Yasushi Ogasawara, Yasuharu Satoh, Chitose Maruyama, Yoshimitsu Hamano, Tohru Dairi
ACS chemical biology, 15, 3, 651, 656, 20 Mar. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Marine microorganisms de novo biosynthesize polyunsaturated fatty acids such as docosahexaenoic acid and eicosapentaenoic acid by polyunsaturated fatty acid (PUFA) synthases composed of three or four polypeptides in a manner similar to fatty acid synthases (FASs). FASs usually possess thioesterase (TE) domains to release free fatty acids from acyl carrier protein (ACP)-tethered intermediates. Here, we investigated the off-loading mechanism with microalgal and bacterial PUFA synthases through in vivo and in vitro experiments. The in vitro experiments with acyltransferase (AT)-like domains and acyl-ACP substrates clearly demonstrated that the AT-like domains catalyzed the hydrolysis of acyl-ACPs to yield free fatty acids.

Subtle Control of Carbon Chain Length in Polyunsaturated Fatty Acid Synthases.
Naka M, Ikeuchi K, Hayashi S, Satoh Y, Ogasawara Y, Dairi T
ACS chemical biology, Nov. 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]

Identification of actinomycin D as a specific inhibitor of the alternative pathway of peptidoglycan biosynthesis.
Ogasawara Y, Shimizu Y, Sato Y, Yoneda T, Inokuma Y, Dairi T
The Journal of antibiotics, 73, 2, 125, 127, Oct. 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Gram-scale fermentative production of ergothioneine driven by overproduction of cysteine in Escherichia coli
Tanaka, Naoyuki, Kawano, Yusuke, Satoh, Yasuharu, Dairi, Tohru, Ohtsu, Iwao
SCIENTIFIC REPORTS, 9, 1895, Feb. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal

In vitro characterization of MitE and MitB: formation of N-acetylglucosaminyl-3-amino-5-hydroxybenzoyl-MmcB as a key intermediate in the biosynthesis of antitumor antibiotic mitomycins.　　　　　　　　　　　　　　　
Y. Ogasawara, Y. Nakagawa, C. Maruyama, Y. Hamano, T. Dairi
Bioorg. Med. Chem. Lett., 29, 2076, 2078, 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Involvement of Peptide Epimerization in Poly-γ-glutamic Acid Biosynthesis.　　　　　　　　　　　　　　　
Y. Ogasawara, M. Shigematsu, S. Sato, H. Kato, T. Dairi
Org. Lett., 21, 3972, 3975, 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Amino Acid Residues Recognizing Isomeric Glutamate Substrates in UDP-N-acetylmuramic acid-L-alanine-glutamate Synthetases.　　　　　　　　　　　　　　　
R. Feng, Y. Satoh, H. Morita, Y. Ogasawara, T. Dairi
ACS Chem. Biol, 14, 975, 978, 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Control mechanism for carbon chain length in polyunsaturated fatty acid synthases.　　　　　　　　　　　　　　　
S. Hayashi, M. Naka, K. Ikeuchi, M. Otsuka, K. Kobayashi, Y. Satoh, Y. Ogasawara, C. Maruyama, Y. Hamano, T. Ujihara, T. Dairi
Angew. Chem. Int. Ed., 58, 6605, 6610, 2019, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Control mechanism for cis-double bond formation by polyunsaturated fatty acid synthases.　　　　　　　　　　　　　　　
S. Hayashi, Y. Satoh, Y. Ogasawara, C. Maruyama, Y. Hamano, T. Ujihara, T. Dairi
Angew. Chem. Int. Ed., 58, 2326, 2330, 2019, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Searching for potent and specific antibiotics against pathogenic Helicobacter and Campylobacter strains.　　　　　　　　　　　　　　　
Y. Ogasawara, T. Dairi
J. Ind. Microbiol. Biotechnol, 46, 409, 414, 2019, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Enzymatic Formation of a Skipped Methyl-Substituted Octaprenyl Side Chain of Longestin (KS-505a): Involvement of Homo-IPP as a Common Extender Unit.
Ozaki T, Shinde SS, Gao L, Okuizumi R, Liu C, Ogasawara Y, Lei X, Dairi T, Minami A, Oikawa H
Angewandte Chemie (International ed. in English), 57, 22, 6629, 6632, May 2018, [Peer-reviewed]

Peptide epimerization machineries found in microorganisms
Yasushi Ogasawara, Tohru Dairi
Frontiers in Microbiology, 9, 156, Frontiers Media S.A., 06 Feb. 2018, [Peer-reviewed], [Invited], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Heterologous and High Production of Ergothioneine in Escherichia coli
Osawa, Ryo, Kamide, Tomoyuki, Satoh, Yasuharu, Kawano, Yusuke, Ohtsu, Iwao, Dairi, Tohru
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 66, 5, 1191, 1196, Feb. 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Functional analysis of methyltransferases participating in streptothricin-related antibiotic biosynthesis
Haruka Niikura, Chitose Maruyama, Yasushi Ogasawara, Kazuo Shin-ya, Tohru Dairi, Yoshimitsu Hamano
Journal of Bioscience and Bioengineering, 125, 2, 148, 154, Elsevier B.V., 01 Feb. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal

Ergothioneine production with Aspergillus oryzae.　　　　　　　　　　　　　　　
S. Takusagawa, Y. Satoh, I. Ohtsu, T. Dairi
Biosci Biotechnol Biochem., 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Aplasmomycin and boromycin are specific inhibitors of the futalosine pathway.　　　　　　　　　　　　　　　
Y. Shimizu, Y. Ogasawara, A. Matsumoto, T. Dairi
J Antibiot, 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Enzymatic formation of a skipped methyl-substituted octaprenyl side chain of longestin (KS-505a): Involvement of homo-IPP as a common extender unit.　　　　　　　　　　　　　　　
T. Ozaki, S.S. Shinde, L. Gao, R. Okuizumi, C. Liu, Y. Ogasawara, X Lei, T. Dairi, A. Minami, H. Oikawa
Angew. Chem. Int. Ed., 20, 6178, 6182, 2018, [Peer-reviewed], ［Internationally co-authored］, [International Magazine]
English, Scientific journal

Biosynthetic Gene Cluster of a D‐Tryptophan‐Containing Lasso Peptide, MS‐271.　　　　　　　　　　　　　　　
Z. Feng, Y. Ogasawara, S. Nomura, T. Dairi
ChemBioChem, 19, 2045, 2048, 2018, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal

Total Biosynthesis of Brassicicenes: Identification of a Key Enzyme for Skeletal Diversification.　　　　　　　　　　　　　　　
A. Tazawa, Y. Ye, T. Ozaki, C. Liu, Y. Ogasawara, T. Dairi, Y. Higuchi, N. Kato, K. Gomi, A. Minami, H. Oikawa
Org. Lett., 20, 6178, 6182, 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal

Novel enzymology in futalosine-dependent menaquinone biosynthesis.　　　　　　　　　　　　　　　
S. Joshi, D. Fedoseyenko, N. Mahanta, H. Manion, S. Naseem, T. Dairi, TP. Begley
Curr Opin Chem Biol, 47, 134, 141, 2018, [Peer-reviewed], ［Internationally co-authored］, [International Magazine]
English, Scientific journal

Synthesis of Acylborons by Ozonolysis of Alkenylboronates: Preparation of an Enantioenriched Amino Acid Acylboronate
Jumpei Taguchi, Toshiki Ikeda, Rina Takahashi, Ikuo Sasaki, Yasushi Ogasawara, Tohru Dairi, Naoya Kato, Yasunori Yamamoto, Jeffrey W. Bode, Hajime Ito
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 56, 44, 13847, 13851, Oct. 2017, [Peer-reviewed]
English, Scientific journal

Biosynthesis of Oligopeptides Using ATP-Grasp Enzymes
Yasushi Ogasawara, Tohru Dairi
CHEMISTRY-A EUROPEAN JOURNAL, 23, 45, 10714, 10724, Aug. 2017, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

N-Phenylacetylation and Nonribosomal Peptide Synthetases with Substrate Promiscuity for Biosynthesis of Heptapeptide Variants, JBIR-78 and JBIR-95
Kunpei Takeda, Kohei Kemmoku, Yasuharu Satoh, Yasushi Ogasawara, Kazuo Shin-ya, Tohru Dairi
ACS CHEMICAL BIOLOGY, 12, 7, 1813, 1819, Jul. 2017, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Identification of tirandamycins as specific inhibitors of the futalosine pathway
Yasushi Ogasawara, Kensuke Kondo, Ayumi Ikeda, Rikako Harada, Tohru Dairi
JOURNAL OF ANTIBIOTICS, 70, 6, 798, 800, Jun. 2017, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

A Glycopeptidyl-Glutamate Epimerase for Bacterial Peptidoglycan Biosynthesis
Ruoyin Feng, Yasuharu Satoh, Yasushi Ogasawara, Tohru Yoshimura, Tohru Dairi
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 139, 12, 4243, 4245, Mar. 2017, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Biosynthesis of the Carbonylmethylene Structure Found in the Ketomemicin Class of Pseudotripeptides
Junpei Kawata, Taiki Naoe, Yasushi Ogasawara, Tohru Dairi
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 56, 8, 2026, 2029, Feb. 2017, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Enhanced production of polyunsaturated fatty acids by enzyme engineering of tandem acyl carrier proteins
Shohei Hayashi, Yasuharu Satoh, Tetsuro Ujihara, Yusuke Takata, Tohru Dairi
SCIENTIFIC REPORTS, 6, 35441, Oct. 2016, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Biosynthesis of Shearinine: Diversification of a Tandem Prenyl Moiety of Fungal Indole Diterpenes
Chengwei Liu, Atsushi Minami, Tohru Dairi, Katsuya Gomi, Barry Scott, Hideaki Oikawa
ORGANIC LETTERS, 18, 19, 5026, 5029, Oct. 2016, [Peer-reviewed]
English, Scientific journal

Characterization of three amidinotransferases involved in the biosynthesis of ketomemicins
Yasushi Ogasawara, Michiko Fujimori, Junpei Kawata, Tohru Dairi
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 26, 15, 3662, 3664, Aug. 2016, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Exploring Peptide Ligase Orthologs in Actinobacteria-Discovery of Pseudopeptide Natural Products, Ketomemicins
Yasushi Ogasawara, Junpei Kawata, Motoyoshi Noike, Yasuharu Satoh, Kazuo Furihata, Tohru Dairi
ACS CHEMICAL BIOLOGY, 11, 6, 1686, 1692, Jun. 2016, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Advanced functionalization of polyhydroxyalkanoate via the UV-initiated thiol-ene click reaction
Kenji Tajima, Kosuke Iwamoto, Yasuharu Satoh, Ryosuke Sakai, Toshifumi Satoh, Tohru Dairi
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 100, 10, 4375, 4383, May 2016, [Peer-reviewed]
English, Scientific journal

Structure and activity relationships of the anti-Mycobacterium antibiotics resorcinomycin and pheganomycin
Yasushi Ogasawara, Koichi Ooya, Michiko Fujimori, Motoyoshi Noike, Tohru Dairi
JOURNAL OF ANTIBIOTICS, 69, 2, 119, 120, Feb. 2016, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Ergothioneine protects Streptomyces coelicolor A3(2) from oxidative stresses
Shunsuke Nakajima, Yasuharu Satoh, Kentaro Yanashima, Tomomi Matsui, Tohru Dairi
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 120, 3, 294, 298, Sep. 2015, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Identification and analysis of the resorcinomycin biosynthetic gene cluster
Koichi Ooya, Yasushi Ogasawara, Motoyoshi Noike, Tohru Dairi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 79, 11, 1833, 1837, 2015, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

A peptide ligase and the ribosome cooperate to synthesize the peptide pheganomycin
Motoyoshi Noike, Takashi Matsui, Koichi Ooya, Ikuo Sasaki, Shouta Ohtaki, Yoshimitsu Hamano, Chitose Maruyama, Jun Ishikawa, Yasuharu Satoh, Hajime Ito, Hiroyuki Morita, Tohru Dairi
NATURE CHEMICAL BIOLOGY, 11, 1, 71, +, Jan. 2015, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

New gene responsible for para-aminobenzoate biosynthesis
Yasuharu Satoh, Masahiro Kuratsu, Daiki Kobayashi, Tohru Dairi
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 117, 2, 178, 183, Feb. 2014, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

A fungal prenyltransferase catalyzes the regular di-prenylation at positions 20 and 21 of paxilline
Chengwei Liu, Motoyoshi Noike, Atsushi Minami, Hideaki Oikawa, Tohru Dairi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 78, 3, 448, 454, 2014, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Functional analysis of a prenyltransferase gene (paxD) in the paxilline biosynthetic gene cluster
Chengwei Liu, Motoyoshi Noike, Atsushi Minami, Hideaki Oikawa, Tohru Dairi
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 98, 1, 199, 206, Jan. 2014, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Rapid Reconstitution of Biosynthetic Machinery for Fungal Metabolites in Aspergillus oryzae: Total Biosynthesis of Aflatrem
Koichi Tagami, Atsushi Minami, Ryuya Fujii, Chengwei Liu, Mizuki Tanaka, Katsuya Gomi, Tohru Dairi, Hideaki Oikawa
Chembiochem, 15, 14, 2076, 2080, 2014, [Peer-reviewed]

Regiospecificities and prenylation mode specificities of the fungal indole diterpene prenyltransferases AtmD and PaxD
Chengwei Liu, Atsushi Minami, Motoyoshi Noike, Hiroaki Toshima, Hideaki Oikawa, Tohru Dairi
Applied and Environmental Microbiology, 79, 23, 7298, 7304, Dec. 2013, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Menaquinone Biosynthesis: Formation of Aminofutalosine Requires a Unique Radical SAM Enzyme
Nilkamal Mahanta, Dmytro Fedoseyenko, Tohru Dairi, Tadhg P. Begley
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 135, 41, 15318, 15321, Oct. 2013, [Peer-reviewed]
English, Scientific journal

In Vitro Reconstitution of the Radical S-Adenosylmethionine Enzyme MqnC Involved in the Biosynthesis of Futalosine-Derived Menaquinone
Lisa E. Cooper, Dmytro Fedoseyenko, Sameh H. Abdelwahed, Soong-Hyun Kim, Tohru Dairi, Tadhg P. Begley
BIOCHEMISTRY, 52, 27, 4592, 4594, Jul. 2013, [Peer-reviewed]
English, Scientific journal

Cellulose complementing factor (Ccp) is a new member of the cellulose synthase complex (terminal complex) in Acetobacter xylinum
Naoki Sunagawa, Takaaki Fujiwara, Takanori Yoda, Shin Kawano, Yasuharu Satoh, Min Yao, Kenji Tajima, Tohru Dairi
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 115, 6, 607, 612, Jun. 2013, [Peer-reviewed]
English, Scientific journal

Reconstitution of Biosynthetic Machinery for Indole-Diterpene Paxilline in Aspergillus oryzae
Koichi Tagami, Chengwei Liu, Atsushi Minami, Motoyoshi Noike, Tetsuya Isaka, Shuhei Fueki, Yoshihiro Shichijo, Hiroaki Toshima, Katsuya Gomi, Tohru Dairi, Hideaki Oikawa
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 135, 4, 1260, 1263, Jan. 2013, [Peer-reviewed]
English, Scientific journal

Cellulose production by Enterobacter sp CJF-002 and identification of genes for cellulose biosynthesis
Naoki Sunagawa, Kenji Tajima, Mariko Hosoda, Shin Kawano, Ryota Kose, Yasuharu Satoh, Min Yao, Tohru Dairi
CELLULOSE, 19, 6, 1989, 2001, Dec. 2012, [Peer-reviewed]
English, Scientific journal

Molecular Breeding of a Fungus Producing a Precursor Diterpene Suitable for Semi-Synthesis by Dissection of the Biosynthetic Machinery
Motoyoshi Noike, Yusuke Ono, Yuji Araki, Ryo Tanio, Yusuke Higuchi, Hajime Nitta, Yoshimitsu Hamano, Tomonobu Toyomasu, Takeshi Sassa, Nobuo Kato, Tohru Dairi
PLOS ONE, 7, 8, e42090, Aug. 2012, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

An Enzyme Catalyzing O-Prenylation of the Glucose Moiety of Fusicoccin A, a Diterpene Glucoside Produced by the Fungus Phomopsis amygdali
Motoyoshi Noike, Chengwei Liu, Yusuke Ono, Yoshimitsu Hamano, Tomonobu Toyomasu, Takeshi Sassa, Nobuo Kato, Tohru Dairi
CHEMBIOCHEM, 13, 4, 566, 573, Mar. 2012, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Menaquinone Biosyntheses in Microorganisms
Tohru Dairi
NATURAL PRODUCT BIOSYNTHESIS BY MICROORGANISMS AND PLANT, PT A, 515, 107, 122, 2012, [Peer-reviewed]
English, In book

Chemo-enzymatic synthesis of polyhydroxyalkanoate (PHA) incorporating 2-hydroxybutyrate by wild-type class I PHA synthase from Ralstonia eutropha
Xuerong Han, Yasuharu Satoh, Toshifumi Satoh, Ken&apos, ichiro Matsumoto, Toyoji Kakuchi, Seiichi Taguchi, Tohru Dairi, Masanobu Munekata, Kenji Tajima
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 92, 3, 509, 517, Nov. 2011, [Peer-reviewed]
English, Scientific journal

Isolation of a thermotolerant bacterium producing medium-chain-length polyhydroxyalkanoate
Y. Satoh, K. Tajima, S. Nakamoto, H. Xuerong, T. Matsushima, T. Ohshima, S. Kawano, T. Erata, T. Dairi, M. Munekata
JOURNAL OF APPLIED MICROBIOLOGY, 111, 4, 811, 817, Oct. 2011, [Peer-reviewed]
English, Scientific journal

20 Analysis of biosynthetic machinery of fusicoccin produced by Phomopsis amygdali(Oral Presentation)
Noike Motoyoshi, Ono Yusuke, Araki Yuji, Tanio Ryo, Hamano Yoshimitsu, Higuchi Yusuke, Toyomasu Tomonobu, Sassa Takeshi, Kato Nobuo, Dairi Tohru
Symposium on the Chemistry of Natural Products, symposium papers, 53, 115, 120, Symposium on the chemistry of natural products, 02 Sep. 2011
Japanese, Both fusicoccin A (FC) and structurally related cotylenin A (CN) are diterpene glucosides and show a phytohormone-like activity. However, only CN induces the differentiation of human myeloid leukemia cells. Since the CN producer lost its ability to proliferate during preservation, a study on the relationship between structure and activity was carried out and an elimination of hydroxyl group at 12-position of FC was essential to have the CN-like activity. Moreover, modified FC with hydroxyl group at 3-position was recently shown to be more effective. Therefore, we tried to identify a gene catalyzing 12-hydroxylation and to breed a mutant producing a modified FC without hydroxyl group at 12-position by a disruption of the gene. Previous identification of fusicocca-2,10(14)-diene synthase gene in Phomopsis amygdali, a FC producer, enabled us to identify a partial gene cluster for bisoynthesis of FC. However, other biosynthetic genes still remained unknown. In this study, we identified another gene cluster containing nine genes by draft genome sequencing. Of these, two cytochrome P450s catalyzing 813 and 9-hydrozylations, glycosylation, methylation, prenylation, and acetylation genes were confirmed to encode enzymes with the expected activities. We also identified a cytochrome P450 catalyzing 12-hydroxylations by a gene disruption experiment. The remaining one cytochrome P450 gene therefore probably catalyzes hydroxylation at the 19-position.

Synthesis of (+/-)-cyclic dehypoxanthine futalosine, the biosynthetic intermediate in an alternative biosynthetic pathway for menaquinones
Arata Yajima, Saki Kouno, Tohru Dairi, Manami Mogi, Ryo Katsuta, Haruo Seto, Tomoo Nukada
TETRAHEDRON LETTERS, 52, 38, 4934, 4937, Sep. 2011, [Peer-reviewed]
English, Scientific journal

Reveromycin A biosynthesis uses RevG and RevJ for stereospecific spiroacetal formation
Shunji Takahashi, Atsushi Toyoda, Yasuyo Sekiyama, Hiroshi Takagi, Toshihiko Nogawa, Masakazu Uramoto, Ryuichiro Suzuki, Hiroyuki Koshino, Takuto Kumano, Suresh Panthee, Tohru Dairi, Jun Ishikawa, Haruo Ikeda, Yoshiyuki Sakaki, Hiroyuki Osada
NATURE CHEMICAL BIOLOGY, 7, 7, 461, 468, Jul. 2011, [Peer-reviewed]
English, Scientific journal

Dioxygenases, Key Enzymes to Determine the Aglycon Structures of Fusicoccin and Brassicicene, Diterpene Compounds Produced by Fungi
Yusuke Ono, Atsushi Minami, Motoyoshi Noike, Yusuke Higuchi, Tomonobu Toyomasu, Takeshi Sassa, Nobuo Kato, Tohru Dairi
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 133, 8, 2548, 2555, Mar. 2011, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Diversity of the Early Step of the Futalosine Pathway
Chisato Arakawa, Masahiro Kuratsu, Kazuo Furihata, Tomoshige Hiratsuka, Nobuya Itoh, Haruo Seto, Tohru Dairi
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 55, 2, 913, 916, Feb. 2011, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Branched fatty acids inhibit the biosynthesis of menaquinone in Helicobacter pylori
Runi Tanaka, Takao Kunisada, Nobuaki Kushida, Keiko Yamada, Shunsuke Ikeda, Motoyoshi Noike, Yuusuke Ono, Nobuya Itoh, Hideto Takami, Haruo Seto, Tohru Dairi
JOURNAL OF ANTIBIOTICS, 64, 1, 151, 153, Jan. 2011, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Convergent strategies in biosynthesis
Tohru Dairi, Tomohisa Kuzuyama, Makoto Nishiyama, Isao Fujii
NATURAL PRODUCT REPORTS, 28, 6, 1054, 1086, 2011, [Peer-reviewed], [Corresponding author]
English

A NEW MENAQUINONE BIOSYNTHETIC PATHWAY, DIVERSITY OF THE EARLY STEP OF THE PATHWAY
Tohru Dairi
PROGRESS ON POST-GENOME TECHNOLOGIES AND MODERN NATURAL PRODUCTS, 2011, 9, 9, 2011, [Peer-reviewed]
English, International conference proceedings

Analysis of the Lactobacillus Metabolic Pathway
Masahiro Kuratsu, Yoshimitsu Hamano, Tohru Dairi
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 76, 21, 7299, 7301, Nov. 2010, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Substrate specificity of the CYC2 enzyme from Kitasatospora griseola: production of sclarene, biformene and novel bicyclic diterpenes by the enzymatic reactions of labdane- and halimane-type diterpene diphosphates
Nakano Chiaki, Hoshino Tsutomu, Sato Tsutomu, Toyomasu Tomonobu, Dairi Tohru, Sassa Takeshi
TETRAHEDRON LETTERS, 51, 1, 125, 128, 06 Jan. 2010, [Peer-reviewed]

Functional analyses of cytochrome P450 genes responsible for the early steps of brassicicene C biosynthesis
Makoto Hashimoto, Yusuke Higuchi, Shunji Takahashi, Hiroyuki Osada, Toshiyuki Sakaki, Tomonobu Toyomasu, Takeshi Sassa, Nobuo Kato, Tohru Dairi
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 19, 19, 5640, 5643, Oct. 2009, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

An alternative menaquinone biosynthetic pathway operating in microorganisms: an attractive target for drug discovery to pathogenic Helicobacter and Chlamydia strains
Tohru Dairi
JOURNAL OF ANTIBIOTICS, 62, 7, 347, 352, Jul. 2009, [Peer-reviewed], [Corresponding author]
English

Enzymatic Properties of Futalosine Hydrolase, an Enzyme Essential to a Newly Identified Menaquinone Biosynthetic Pathway
Tomoshige Hiratsuka, Nobuya Itoh, Haruo Seto, Tohru Dairi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 73, 5, 1137, 1141, May 2009, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Identification and functional analysis of brassicicene C biosynthetic gene cluster in Alternaria brassicicola
Atsushi Minami, Naoto Tajima, Yusuke Higuchi, Tomonobu Toyomasu, Takeshi Sassa, Nobuo Kato, Tohru Dairi
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 19, 3, 870, 874, Feb. 2009, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Biosynthetic Gene-Based Secondary Metabolite Screening: A New Diterpene, Methyl Phomopsenonate, from the Fungus Phomopsis amygdali
Tomonobu Toyomasu, Akane Kaneko, Tetsuo Tokiwano, Yuya Kanno, Yuri Kanno, Rie Niida, Shigeyoshi Miura, Taiki Nishioka, Chiho Ikeda, Wataru Mitsuhashi, Tohru Dairi, Tomikazu Kawano, Hideaki Oikawa, Nobuo Kato, Takeshi Sassa
JOURNAL OF ORGANIC CHEMISTRY, 74, 4, 1541, 1548, Feb. 2009, [Peer-reviewed]
English, Scientific journal

An alternative menaquinone biosynthetic pathway operating in microorganisms
Tohru Dairi
Seikagaku, 81, 2, 95, 99, 2, 2009, [Peer-reviewed], [Corresponding author]
Japanese

An alternative menaquinone biosynthetic pathway operating in microorganisms
Tomoshige Hiratsuka, Kazuo Furihata, Jun Ishikawa, Haruyuki Yamashita, Nobuya Itoh, Haruo Seto, Tohru Dairi
SCIENCE, 321, 5896, 1670, 1673, Sep. 2008, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Identification of diterpene biosynthetic gene clusters and functional analysis of labdane-related diterpene cyclases in Phomopsis amygdali
Tomonobu Toyomasu, Rie Nuda, Hiromichi Kenmoku, Yuri Kanno, Shigeyoshi Miura, Chiaki Nakano, Yoshihito Shiono, Wataru Mitsuhashi, Hiroaki Toshima, Hideaki Oikawa, Tsutomu Hoshino, Tohru Dairi, Nobuo Kato, Takeshi Sassa
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 72, 4, 1038, 1047, Apr. 2008, [Peer-reviewed]
English, Scientific journal

Studies on a new biosynthetic pathway for menaquinone
Haruo Seto, Yuusuke Jinnai, Tomoshige Hiratsuka, Miwako Fukawa, Kazuo Furihata, Nobuya Itoh, Tohru Dairi
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 130, 17, 5614, +, Apr. 2008, [Peer-reviewed], [Last author]
English, Scientific journal

Cloning of the gene cluster responsible for the biosynthesis of brasilicardin A, a unique diterpenoid
Yutaka Hayashi, Nobuyasu Matsuura, Hiroaki Toshima, Nobuya Itoh, Jun Ishikawa, Yuzuru Mikami, Tohru Dairi
JOURNAL OF ANTIBIOTICS, 61, 3, 164, 174, Mar. 2008, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Comparison of the enzymatic properties of ent-copalyl diphosphate synthases in the biosynthesis of phytoalexins and gibberellins in rice
Yutaka Hayashi, Tomonobu Toyomasu, Yuko Hirose, Yu Onodera, Wataru Mitsuhashi, Hisakazu Yamane, Takeshi Sassa, Tohru Dairi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 72, 2, 523, 530, Feb. 2008, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Engineering the phenylacetaldehyde reductase mutant for improved substrate conversion in the presence of concentrated 2-propanol
Yoshihide Makino, Tohru Dairi, Nobuya Itoh
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 77, 4, 833, 843, Dec. 2007, [Peer-reviewed]
English, Scientific journal

Cloning of the gene cluster responsible for biosynthesis of KS-505a (longestin), a unique tetraterpenoid
Yutaka Hayashi, Hiroyasu Onaka, Nobuya Itoh, Haruo Seto, Tohru Dairi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 12, 3072, 3081, Dec. 2007, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Studies on terpenoids produced by actinomycetes: Oxaloterpins A, B, C, D, and E, Diterpenes from Streptomyces sp KO-3988
Keiichiro Motohashi, Ryoko Ueno, Masayuki Sue, Kazuo Furihata, Takuo Matsumoto, Tohru Dairi, Satoshi Omura, Haruo Seto
JOURNAL OF NATURAL PRODUCTS, 70, 11, 1712, 1717, Nov. 2007, [Peer-reviewed]
English, Scientific journal

Fusicoccins are biosynthesized by an unusual chimera diterpene synthase in fungi
Tomonobu Toyomasu, Mai Tsukahara, Akane Kaneko, Rie Niida, Wataru Mitsuhashi, Tohru Dairi, Nobuo Kato, Takeshi Sassa
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 104, 9, 3084, 3088, Feb. 2007, [Peer-reviewed]
English, Scientific journal

Functional analysis of eubacterial ent-copalyl diphosphate synthase and pimara-9(11),15-diene synthase with unique primary sequences
Chiho Ikeda, Yutaka Hayashi, Nobuya Itoh, Haruo Seto, Tohru Dairi
JOURNAL OF BIOCHEMISTRY, 141, 1, 37, 45, Jan. 2007, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

A comparative analysis of the sugar phosphate cyclase superfamily involved in primary and secondary metabolism
Xiumei Wu, Patricia M. Flatt, Oliver Schloerke, Axel Zeeck, Tohru Dairi, Taifo Mahmud
CHEMBIOCHEM, 8, 2, 239, 248, Jan. 2007, [Peer-reviewed]
English, Scientific journal

P-355 Biosynthesis of a Natural Polyketide-Isoprenoid Hybrid Compound Furaquinocin A : Identification and Heterologous Expression of the Gene Cluster
Hayashi Yutaka, Kawasaki Takashi, Kuzuyama Tomohisa, Furihata Kazuo, Itoh Nobuya, Seto Haruo, Dairi Tohru
International Symposium on the Chemistry of Natural Products, 2006, "P, 355", Symposium on the chemistry of natural products, 23 Jul. 2006
English

Biosynthesis of a natural polyketide-isoprenoid hybrid compound, furaquinocin A: Identification and heterologous expression of the gene cluster
T Kawasaki, Y Hayashi, T Kuzuyama, K Furihata, N Itoh, H Seto, T Dairi
JOURNAL OF BACTERIOLOGY, 188, 4, 1236, 1244, Feb. 2006, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Gene cloning and expression of Leifsonia alcohol dehydrogenase (LSADH) involved in asymmetric hydrogen-transfer bioreduction to produce (R)-form chiral alcohols
K Inoue, Y Makino, T Dairi, N Itoh
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70, 2, 418, 426, Feb. 2006, [Peer-reviewed]
English, Scientific journal

Engineering of phenylacetaldehyde reductase for efficient substrate conversion in concentrated 2-propanol
Y Makino, K Inoue, T Dairi, N Itoh
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 71, 8, 4713, 4720, Aug. 2005, [Peer-reviewed]
English, Scientific journal

fldA is an essential gene required in the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis
KJ Puan, H Wang, T Dairi, T Kuzuyama, CT Morita
FEBS LETTERS, 579, 17, 3802, 3806, Jul. 2005, [Peer-reviewed]
English, Scientific journal

Studies on biosynthetic genes and enzymes of isoprenoids produced by actinomycetes
T Dairi
JOURNAL OF ANTIBIOTICS, 58, 4, 227, 243, Apr. 2005, [Peer-reviewed], [Corresponding author]
English

Mycobacterium tuberculosis H37Rv3377c encodes the diterpene cyclase for producing the halimane skeleton
C Nakano, T Okamura, T Sato, T Dairi, T Hoshino
CHEMICAL COMMUNICATIONS, Issue 8: 1016–1018, 8, 1016, 1018, Feb. 2005, [Peer-reviewed]
English, Scientific journal

Purification and characterization of NADPH-dependent aldo-keto reductase specific for beta-keto esters from Penicillium citrinum, and production of methyl (S)-4-bromo-3-hydroxybutyrate
N Itoh, H Asako, K Banno, Y Makino, M Shinohara, T Dairi, R Wakita, M Shimizu
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 66, 1, 53, 62, Nov. 2004, [Peer-reviewed]
English, Scientific journal

Presence of copalyl diphosphate synthase gene in an actinomycete possessing the mevalonate pathway
T Kawasaki, T Kuzuyama, Y Kuwamori, N Matsuura, N Itoh, K Furihata, H Seto, T Dairi
JOURNAL OF ANTIBIOTICS, 57, 11, 739, 747, Nov. 2004, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Identification and cloning of the gene involved in the final step of chlortetracycline biosynthesis in Streptomyces aureofaciens
T Nakano, K Miyake, H Endo, T Dairi, T Mizukami, R Katsumata
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68, 6, 1345, 1352, Jun. 2004, [Peer-reviewed]
English, Scientific journal

Heterologous mevalonate production in Streptawyces lividans TK23
T Kuzuyama, T Dairi, H Yamashita, Y Shoji, H Seto
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68, 4, 931, 934, Apr. 2004, [Peer-reviewed]
English, Scientific journal

3P110 Engineering of phenylacetaldehyde reductase for improved substrate conversion in concentrated 2-propanol
Makino Y., Dairi T., Itoh N.
Seibutsu Butsuri, 44, S217, The Biophysical Society of Japan General Incorporated Association, 2004
Japanese

A relationship between the mevalonate pathway and isoprenoid production in actinomycetes
T Kawasaki, T Kuzuyama, K Furihata, N Itoh, H Seto, T Dairi
JOURNAL OF ANTIBIOTICS, 56, 11, 957, 966, Nov. 2003, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

A novel enzyme, D-3-hydroxyaspartate aldolase from Paracoccus denitrificans IFO 13301: purification, characterization, and gene cloning
JQ Liu, T Dairi, N Itoh, M Kataoka, S Shimizu
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 62, 1, 53, 60, Jul. 2003, [Peer-reviewed]
English, Scientific journal

A new approach for the investigation of isoprenoid biosynthesis featuring pathway switching, deuterium hyperlabeling, and H-1 NMR spectroscopy. The reaction mechanism of a novel Streptomyces diterpene cyclase
T Eguchi, Y Dekishima, Y Hamano, T Dairi, H Seto, K Kakinuma
JOURNAL OF ORGANIC CHEMISTRY, 68, 14, 5433, 5438, Jul. 2003, [Peer-reviewed]
English, Scientific journal

Interconversion of the product specificity of type I eubacterial farnesyl diphosphate synthase and geranylgeranyl diphosphate synthase through one amino acid substitution
T Kawasaki, Y Hamano, T Kuzuyama, N Itoh, H Seto, T Dairi
JOURNAL OF BIOCHEMISTRY, 133, 1, 83, 91, Jan. 2003, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Functional analysis of eubacterial diterpene cyclases responsible for biosynthesis of a diterpene antibiotic, terpentecin
Y Hamano, T Kuzuyama, N Itoh, K Furihata, H Seto, T Dairi
JOURNAL OF BIOLOGICAL CHEMISTRY, 277, 40, 37098, 37104, Oct. 2002, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Detection of the mevalonate pathway in streptomyces species using the 3-hydroxy-3-methylglutaryl coenzyme A reductase gene
T Kuzuyama, S Takahashi, T Dairi, H Seto
JOURNAL OF ANTIBIOTICS, 55, 10, 919, 923, Oct. 2002, [Peer-reviewed]
English, Scientific journal

Chiral alcohol production by NADH-dependent phenylacetaldehyde reductase coupled with in situ regeneration of NADH
N Itoh, M Matsuda, M Mabuchi, T Dairi, J Wang
EUROPEAN JOURNAL OF BIOCHEMISTRY, 269, 9, 2394, 2402, May 2002, [Peer-reviewed]
English, Scientific journal

Growth-phase dependent expression of the mevalonate pathway in a terpenoid antibiotic-producing Streptomyces strain
Y Hamano, T Dairi, M Yamamoto, T Kuzuyama, N Itoh, H Seto
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66, 4, 808, 819, Apr. 2002, [Peer-reviewed], [Last author, Corresponding author]
English, Scientific journal

Reaction mechanism of the Co2+-activated multifunctional bromoperoxidase-esterase from Pseudomonas putida IF-3
T Kawanami, M Miyakoshi, T Dairi, N Itoh
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 398, 1, 94, 100, Feb. 2002, [Peer-reviewed]
English, Scientific journal

Studies on eubacterial diterpene cyclases found in actinomycetes
T Dairi, T Hamano, N Itoh, T Kuzuyama, K Furihata, H Seto
NIPPON NOGEIKAGAKU KAISHI-JOURNAL OF THE JAPAN SOCIETY FOR BIOSCIENCE BIOTECHNOLOGY AND AGROCHEMISTRY, 76, 12, 1191, 1194, 2002, [Peer-reviewed], [Lead author, Corresponding author]
Japanese, Scientific journal

Eubacterial diterpene cyclase genes essential for production of the isoprenoid antibiotic terpentecin
T Dairi, Y Hamano, T Kuzuyama, N Itoh, K Furihata, H Seto
JOURNAL OF BACTERIOLOGY, 183, 20, 6085, 6094, Oct. 2001, [Peer-reviewed], [Lead author, Corresponding author]
English, Scientific journal

Cloning of a gene cluster encoding enzymes responsible for the mevalonate pathway from a terpenoid-antibiotic-producing Streptomyces strain
Y Hamano, T Dairi, M Yamamoto, T Kawasaki, K Kaneda, T Kuzuyama, N Itoh, H Seto
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 65, 7, 1627, 1635, Jul. 2001, [Peer-reviewed], [Corresponding author]
English, Scientific journal

Cloning and biochemical characterization of Co(2+)-activated bromoperoxidase-esterase (perhydrolase) from Pseudomonas putida IF-3 strain.
Itoh N, Kawanami T, Liu JQ, Dairi T, Miyakoshi M, Nitta C, Kimoto Y
Biochimica et biophysica acta, 1545, 53, 66, 1-2, Feb. 2001, [Peer-reviewed]

2 Development of Methodology for Biochemical and Biosynthetic Studies of Isoprenoid Compounds using Perdeuterated Mevalonate
Eguchi Tadashi, Dekishima Yasumasa, Matsushima Yoshitaka, Tamegai Hideyuki, Kakinuma Katsumi, Takagi Motoki, Kuzuyama Tomohisa, Seto Haruo, Misawa Norihiko, Hamano Yoshimitsu, Dairi Tohru
Symposium on the Chemistry of Natural Products, symposium papers, 43, 7, 12, Symposium on the Chemistry of Natural Products Steering Committee, 2001
Japanese, Isoprenoids are chemically diverse in nature, ubiquitous in living organisms, and crucial in biological processes. The biosynthesis of such isoprenoids proceeds through mevalonate and nonmevalonate pathways depending upon organisms and cellular organella, isopentenyl diphosphate (IPP) being a key intermediate in both cases. Metabolic engineering and control of these pathways should thus provide new opportunity to study intriguing chemistry and biochemistry involved and to develop selective chemotherapeutic agents and isoprenoid-related materials. We describe a new practical approach to the preparation of highly- and multiply deuterated isoprenoids, zeaxanthin and diterpene anitibiotic terpentecin being as examples, and its potential for analyzing the biosynthetic mechanism of isoprenoids, based on the metabolic engineering of microorganisms. Obviously, deuterium-labeled compounds are invaluable in biochemical, bioorganic as well as physicochemical research. Metabolically engineered E. coli DK223 (pTMV20, pACCAR25ΔcrtX) produces zeaxanthin under the presence of mevalonate. Fully deuterated mevalonolactone-d_9 (MVL-d_9), which had been synthesized, was supplemented to the culture of the above triply-engineered E. coli, and the biosynthesized zeaxanthin was extracted and purified by repeated chromatography. All the zeaxanthin formed was proved to be derived only from the supplemented MVL-d_9. This was the first example of such highly and multiply deuterated zeaxanthin, and clearly demonstrated significant potential of the present approach for the preparation of various isotope-labeled isoprenoids. Additional example of this approach was also demonstrated in the mechanistic study of cyclization reaction in the biosynthesis of diterpene anitibiotic terpentecin. Straightforward stereochemical analysis of isoprenoid biosynthesis was demonstrated by one-shot labeling of MVL-d_9 and ^1H NMR spectroscopy. Precise analysis of the simplified proton spectra of highly deuterated isoprenoids, especially under the deuterium decoupled conditions, appeared to be beneficial for mechanistic enzymology, particularly, for the key transformation involving proton attack and proton quench as observed in the terpene cyclase reactions.

Gene cloning and overproduction of low-specificity D-threonine aldolase from Alcaligenes xylosoxidans and its application for production of a key intermediate for parkinsonism drug
JQ Liu, M Odani, T Yasuoka, T Dairi, N Itoh, M Kataoka, S Shimizu, H Yamada
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 54, 1, 44, 51, Jul. 2000, [Peer-reviewed]
English, Scientific journal

Cloning of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from terpenoid antibiotic-producing Streptomyces strains
T Dairi, Y Motohira, T Kuzuyama, S Takahashi, N Itoh, H Seto
MOLECULAR AND GENERAL GENETICS, 262, 6, 957, 964, Jan. 2000, [Peer-reviewed], [Lead author, Corresponding author]
English, Scientific journal

Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding phenylacetaldehyde reductase from styrene-assimilating Corynebacterium sp strain ST-10
JC Wang, M Sakakibara, JQ Liu, T Dairi, N Itoh
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 52, 3, 386, 392, Sep. 1999, [Peer-reviewed]
English, Scientific journal

Development of a self-cloning system for Actinomadura verrucosospora and identification of polyketide synthase genes essential for production of the angucyclic antibiotic pradimicin
T Dairi, Y Hamano, T Furumai, T Oki
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 65, 6, 2703, 2709, Jun. 1999, [Peer-reviewed], [Lead author]
English, Scientific journal

A new route to L-threo-3-[4-(methylthio)phenylserine], a key intermediate for the synthesis of antibiotics: recombinant low-specificity D-threonine aldolase-catalyzed stereospecific resolution
JQ Liu, M Odani, T Dairi, N Itoh, S Shimizu, H Yamada
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 51, 5, 586, 591, May 1999, [Peer-reviewed]
English, Scientific journal

A novel metal-activated pyridoxal enzyme with a unique primary structure, low specificity D-threonine aldolase from Arthrobacter sp. strain DK-38 - Molecular cloning and cofactor characterization
JQ Liu, T Dairi, N Itoh, M Kataoka, S Shimizu, H Yamada
JOURNAL OF BIOLOGICAL CHEMISTRY, 273, 27, 16678, 16685, Jul. 1998, [Peer-reviewed]
English, Scientific journal

Gene cloning, biochemical characterization and physiological role of a thermostable low-specificity L-threonine aldolase from Escherichia coli
JQ Liu, T Dairi, N Itoh, M Kataoka, S Shimizu, H Yamada
EUROPEAN JOURNAL OF BIOCHEMISTRY, 255, 1, 220, 226, Jul. 1998, [Peer-reviewed]
English, Scientific journal

Gene cloning, nucleotide sequencing, and purification and characterization of the low-specificity L-threonine aldolase from Pseudomonas sp. strain NCIMB 10558
JQ Liu, S Ito, T Dairi, N Itoh, M Kataoka, S Shimizu, H Yamada
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 64, 2, 549, 554, Feb. 1998, [Peer-reviewed]
English, Scientific journal

Shotgun cloning and characterization of the thymidylate synthase-encoding gene from Mycobacterium bovis BCG
S Matsumoto, H Yukitake, N Ohara, T Dairi, H Kanbara, T Yamada
MICROBIOLOGY AND IMMUNOLOGY, 42, 1, 15, 21, 1998, [Peer-reviewed]
English, Scientific journal

Crystallization of Arthrobacter sp. strain 1C N-(1-D-carboxyethyl)-L-norvaline dehydrogenase and its complex with NAD+
Britton KL, Rogers HF, Asano Y, Dairi T, Kato Y, Stillman TJ, Rice DW
Acta crystallographica. Section D, Biological crystallography, 53, 124, 126, Jan. 1998, [Peer-reviewed]
English, Scientific journal

Crystallization of Arthrobacter sp. strain 1C N-(1-D-carboxyethyl)-L-norvaline dehydrogenase and its complex with NAD+
K. Linda Britton, H. Fiona Rogers, Yasuhisa Asano, Tohru Dairi, Yasuo Kato, Timothy J. Stillman, David W. Rice
Acta Crystallographica Section D: Biological Crystallography, 54, 1, 124, 126, Pt 1, 01 Jan. 1998, [Peer-reviewed]
English, Scientific journal

Cloning and nucleotide sequence of the putative polyketide synthase genes for pradimicin biosynthesis from Actinomadura hibisca
T Dairi, Y Hamano, Y Igarashi, T Furumai, T Oki
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 61, 9, 1445, 1453, Sep. 1997, [Peer-reviewed], [Lead author]
English, Scientific journal

L-allo-threonine aldolase from Aeromonas jandaei DK-39: Gene cloning, nucleotide sequencing, and identification of the pyridoxal 5'-phosphate-binding lysine residue by site-directed mutagenesis
JQ Liu, T Dairi, M Kataoka, S Shimizu, H Yamada
JOURNAL OF BACTERIOLOGY, 179, 11, 3555, 3560, Jun. 1997, [Peer-reviewed]
English, Scientific journal

The GLY1 gene of Saccharomyces cerevisiae encodes a low-specific L-threonine aldolase that catalyzes cleavage of L-allo-threonine and L-threonine to glycine - Expression of the gene in Escherichia coli and purification and characterization of the enzyme
JQ Liu, S Nagata, T Dairi, H Misono, S Shimizu, H Yamada
EUROPEAN JOURNAL OF BIOCHEMISTRY, 245, 2, 289, 293, Apr. 1997, [Peer-reviewed]
English, Scientific journal

An alkaline D-stereospecific endopeptidase with beta-lactamase activity from Bacillus cereus
Y Asano, H Ito, T Dairi, Y Kato
JOURNAL OF BIOLOGICAL CHEMISTRY, 271, 47, 30256, 30262, Nov. 1996, [Peer-reviewed]
English, Scientific journal

NUCLEOTIDE-SEQUENCE OF FORTIMICIN KL1 METHYLTRANSFERASE GENE ISOLATED FROM MICROMONOSPORA-OLIVASTEROSPORA, AND COMPARISON OF ITS DEDUCED AMINO-ACID-SEQUENCE WITH THOSE OF METHYLTRANSFERASES INVOLVED IN THE BIOSYNTHESIS OF BIALAPHOS AND FOSFOMYCIN
T KUZUYAMA, T SEKI, T DAIRI, T HIDAKA, H SETO
JOURNAL OF ANTIBIOTICS, 48, 10, 1191, 1193, Oct. 1995, [Peer-reviewed]
English

A SELF-DEFENSE GENE HOMOLOGOUS TO TETRACYCLINE EFFLUXING GENE ESSENTIAL FOR ANTIBIOTIC PRODUCTION IN STREPTOMYCES-AUREOFACIENS
T DAIRI, K AISAKA, R KATSUMATA, M HASEGAWA
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 10, 1835, 1841, Oct. 1995, [Peer-reviewed], [Lead author]
English, Scientific journal

NUCLEOTIDE SEQUENCING OF PHENYLALANINE DEHYDROGENASE GENE FROM BACILLUS-BADIUS-IAM-11059
A YAMADA, T DAIRI, Y OHNO, XL HUANG, Y ASANO
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 10, 1994, 1995, Oct. 1995, [Peer-reviewed]
English

CLONING, NUCLEOTIDE SEQUENCING, AND EXPRESSION OF AN OPINE DEHYDROGENASE GENE FROM ARTHROBACTER SP STRAIN 1C
T DAIRI, Y ASANO
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 61, 8, 3169, 3171, Aug. 1995, [Peer-reviewed], [Lead author]
English

CONSERVED ORGANIZATION OF GENES FOR BIOSYNTHESIS OF CHLORTETRACYCLINE IN STREPTOMYCES STRAINS
T DAIRI, T NAKANO, T MIZUKAMI, K AISAKA, M HASEGAWA, R KATSUMATA
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 7, 1360, 1361, Jul. 1995, [Peer-reviewed], [Lead author]
English

CLONING AND NUCLEOTIDE-SEQUENCE OF THE GENE RESPONSIBLE FOR CHLORINATION OF TETRACYCLINE
T DAIRI, T NAKANO, K AISAKA, R KATSUMATA, M HASEGAWA
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 59, 6, 1099, 1106, Jun. 1995, [Peer-reviewed], [Lead author]
English, Scientific journal

Anti-infectives: Overview pradimicins: Potential antifungal and anti-HIV agents
Toshikazu Oki, Tohru Dairi
Expert Opinion on Therapeutic Patents, 4, 12, 1483, 1491, Informa Healthcare, 01 Dec. 1994
English, Scientific journal

CHARACTERIZATION OF 2 DIFFERENT TYPES OF RESISTANCE GENES AMONG PRODUCERS OF FORTIMICIN-GROUP ANTIBIOTICS
T OHTA, T DAIRI, M HASEGAWA
JOURNAL OF GENERAL MICROBIOLOGY, 139, 139, 591, 599, Mar. 1993, [Peer-reviewed]
English, Scientific journal

ORGANIZATION AND NATURE OF FORTIMICIN-A (ASTROMICIN) BIOSYNTHETIC GENES STUDIED USING A COSMID LIBRARY OF MICROMONOSPORA-OLIVASTEROSPORA DNA
T DAIRI, T OHTA, E HASHIMOTO, M HASEGAWA
MOLECULAR & GENERAL GENETICS, 236, 1, 39, 48, Dec. 1992, [Peer-reviewed], [Lead author]
English, Scientific journal

N-FORMIMIDOYL FORTIMICIN-A SYNTHASE, A UNIQUE OXIDASE INVOLVED IN FORTIMICIN A BIOSYNTHESIS - PURIFICATION, CHARACTERIZATION AND GENE CLONING
T DAIRI, K YAMAGUCHI, M HASEGAWA
MOLECULAR & GENERAL GENETICS, 236, 1, 49, 59, Dec. 1992, [Peer-reviewed], [Lead author]
English, Scientific journal

SELF CLONING IN MICROMONOSPORA-OLIVASTEROSPORA OF FMS GENES FOR FORTIMICIN-A (ASTROMICIN) BIOSYNTHESIS
T DAIRI, T OHTA, E HASHIMOTO, M HASEGAWA
MOLECULAR & GENERAL GENETICS, 232, 2, 262, 270, Mar. 1992, [Peer-reviewed], [Lead author]
English, Scientific journal

A NOVEL, HIGHLY EFFICIENT GENE-CLONING SYSTEM FOR MICROMONOSPORA STRAINS
M HASEGAWA, T DAIRI, T OHTA, E HASHIMOTO
JOURNAL OF BACTERIOLOGY, 173, 21, 7004, 7011, Nov. 1991, [Peer-reviewed]
English, Scientific journal

COMMON BIOSYNTHETIC FEATURE OF FORTIMICIN-GROUP ANTIBIOTICS
T DAIRI, M HASEGAWA
JOURNAL OF ANTIBIOTICS, 42, 6, 934, 943, Jun. 1989, [Peer-reviewed], [Lead author]
English, Scientific journal

POSITIVE CONTROL OF TRANSCRIPTION INITIATION IN ESCHERICHIA-COLI - A BASE SUBSTITUTION AT THE PRIBNOW BOX RENDERS OMPF EXPRESSION INDEPENDENT OF A POSITIVE REGULATOR
T DAIRI, K INOKUCHI, T MIZUNO, S MIZUSHIMA
JOURNAL OF MOLECULAR BIOLOGY, 184, 1, 1, 6, 1985, [Peer-reviewed], [Lead author]
English

Dissecting Ketosynthase Domains in Microalgal Polyunsaturated Fatty Acid Synthase
尾形海斗, 仲間陸, 小林飛悠, 角田毅, 丸山千登勢, 濱野吉十, 氏原哲朗, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Dissecting Dehydratase Domains in Microalgal Polyunsaturated Fatty Acid Synthase
川田智哉, 小林飛悠, 大塚慎, 角田毅, 丸山千登勢, 濱野吉十, 氏原哲朗, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Biosynthetic Study of Antitumor Antibiotic Mitomycins
高橋佑大, 角田毅, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Studies on biosynthesis of hydrazide-containing natural product
西田壮吾, 成冨駿, 小倉勇輝, 山崎陽香, 角田毅, 勝山陽平, 勝山陽平, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Identification of Novel GTP Cyclohydrolase with Genome Mining
梅澤朱理, 角田毅, 新宅一平, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Biosynthetic Study of Proteasome Inhibitor
角田毅, 古村淳吉, 山崎陽香, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Searching for Specific Inhibitors of the Alternative Menaquinone Biosynthetic Pathway
鬼柳春花, 菊池麻里江, 角田毅, 野中健一, 野中健一, 稲橋佑起, 稲橋佑起, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Searching for Specific Inhibitors of the Alternative Peptidoglycan Biosynthetic Pathway
梅津秀平, 角田毅, 鬼柳春花, 稲橋佑起, 稲橋佑起, 野中健一, 野中健一, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2024, 2024

Bioprocess for tyrosol production with bacterial phenylalanine hydroxylase
SHEN Ning, SATOH Yasuharu, OGASAWARA Yasushi, DAIRI Tohru, 日本栄養・食糧学会北海道支部大会講演要旨集, 53rd (CD-ROM), 2023

Engineering Escherichia coli for high production of tyrosol from phenylalanine
SHEN Ning, SATOH Yasuharu, KOMA Daisuke, OGASAWARA Yasushi, DAIRI Tohru, 日本生物工学会大会講演要旨集, 75th, 2023

Searching for Specific Inhibitors of the Alternative Pathway of Peptidoglycan Biosynthesis
梅津秀平, 角田毅, 稲橋佑起, 野中健一, 小笠原泰志, 大利徹, 日本放線菌学会大会講演要旨集, 37th, 2023

Biosynthetic Studies on Antitumor Antibiotic Mitomycins
高橋佑大, 中川陽, 角田毅, 大利徹, 小笠原泰志, 日本放線菌学会大会講演要旨集, 37th, 2023

Biosynthetic studies on natural product containing α,α-disubstituted amino acid
角田毅, 古村淳吉, 山崎陽香, 小笠原泰志, 大利徹, 日本放線菌学会大会講演要旨集, 37th, 2023

Involvement of Peptide Epimerization in the biosynthesis of Linaridin Class Ribosomally Synthesized and Post-translationally Modified Peptides
XIAO Wanlu, OGASAWARA Yasushi, SATOH Yasuharu, DAIRI Tohru, 日本農芸化学会大会講演要旨集(Web), 2022, 2022

Involvement of peptide epimerization in the biosynthesis of the linaridin class ribosomally synthesized and post-translationally modified peptides
XIAO Wanlu, OGASAWARA Yasushi, SATOH Yasuharu, DAIRI Tohru, 日本放線菌学会大会講演要旨集, 36th, 2022

Biosynthesis of lasso peptide RES-701
山谷優花, 小笠原泰志, 佐藤康治, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2022, 2022

Dissecting Biosynthetic Machinery of Microalgal Polyunsaturated Fatty Acid Synthase
仲間陸, 小林飛悠, 大塚慎, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2022, 2022

微細藻類由来DHA合成酵素のdehydrataseドメインの機能解析
小林飛悠, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会東北支部大会プログラム・講演要旨集, 2022, 2022

Biosynthesis of D-amino acid containing peptides in microorganism
小笠原泰志, 佐藤康治, 大利徹, 生化学, 93, 3, 329, 337, Jun. 2021, [Peer-reviewed], [Invited], [Last author, Corresponding author]
Japanese

Analysis of a novel pathway for para-aminobenzoate biosynthesis in bacteria
XIAO Wanlu, SATOH Yasuharu, OGASAWARA Yasushi, DAIRI Tohru, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2021, 2021

微細藻類由来DHA合成酵素の炭素鎖長制御機構の解明
仲間陸, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2021, 2021

微細藻類由来DHA合成酵素のdehydrataseドメインの機能解析
小林飛悠, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2021, 2021

Epimerase involved in polyglutamic acid biosynthesis
加藤陽菜多, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2021, 2021

Biosynthesis of the antitumor antibiotics mitomycins
中川陽, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2021, 2021

Reaction mechanism of ATP-dependent peptide epimerase MurL
中尾優文, 小笠原泰志, 佐藤康治, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2021, 2021

新規ATP依存ペプチドエピメラーゼMurLの反応機構解析
中尾優文, 小笠原泰志, 佐藤康治, 大利徹, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2020, 2020

Enhanced production of ergothioneine by recombinant Escherichia coli
田草川隼, 上出倫敬, 佐藤康治, 小笠原泰志, 大津厳生, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2020, 2020

海洋微生物における多価不飽和脂肪酸生合成酵素の解析と応用　　　　　　　　　　　　　　　
林 祥平, 小笠原泰志, 佐藤康治, 大利 徹, B&I, 77, 6, 448, 452, Nov. 2019, [Invited]
Japanese, Introduction scientific journal

多価不飽和脂肪酸生合成酵素の論理的機能改変
池内健心, 中真以, 林祥平, 大塚慎, 小林洸太, 佐藤康治, 小笠原泰志, 氏原哲朗, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.1D2a12 (WEB ONLY), 05 Mar. 2019
Japanese

in vitro解析による多価不飽和脂肪酸生合成酵素の炭素鎖長制御機構の解明
林祥平, 小笠原泰志, 佐藤康治, 丸山千登勢, 濱野吉十, 氏原哲朗, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.1D2a10 (WEB ONLY), 05 Mar. 2019
Japanese

in vitro解析による多価不飽和脂肪酸生合成酵素のcis二重結合形成機構の解明
林祥平, 小笠原泰志, 佐藤康治, 丸山千登勢, 濱野吉十, 氏原哲朗, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.1D2a09 (WEB ONLY), 05 Mar. 2019
Japanese

精密解析に基づく多価不飽和脂肪酸生合成酵素の機能改変
林祥平, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.3S13D6a5 (WEB ONLY), 05 Mar. 2019
Japanese

in vivo解析による多価不飽和脂肪酸生合成酵素の生成物制御ドメインの同定
林祥平, 中真以, 佐藤康治, 氏原哲朗, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2019, ROMBUNNO.1D2a08 (WEB ONLY), 05 Mar. 2019
Japanese

ロンゲスチン生合成における特異なメチル基導入機構の解明
尾崎太郎, SHINDE Sandip S., GAO Lei, 奥泉諒, LIU Chengwei, 小笠原泰志, 大利徹, 南篤志, 及川英秋, 日本農芸化学会大会講演要旨集(Web), 2019, 2019

抗腫瘍抗生物質マイトマイシンの生合成解析
中川陽, 小笠原泰志, 大利徹, 日本生物工学会大会講演要旨集, 71st, 2019

抗腫瘍抗生物質マイトマイシンの初期生合成機構のin vitro解析
中川陽, 小笠原泰志, 丸山千登勢, 濱野吉十, 大利徹, 日本放線菌学会大会講演要旨集, 34th, 2019

ポリグルタミン酸生合成における新規エピメリ化反応
佐藤将太, 重松真由子, 加藤陽菜多, 小笠原泰志, 大利徹, 日本生物工学会大会講演要旨集, 71st, 2019

精密解析に基づく多価不飽和脂肪酸生合成酵素の機能改変
林祥平, 佐藤康治, 小笠原泰志, 大利徹, 日本生物工学会大会講演要旨集, 71st, 2019

天然抗酸化剤~エルゴチオネイン~の高効率微生物生産
大利徹, アサヒグループ学術振興財団研究紀要(Web), 2019, 2019

多価不飽和脂肪酸合成酵素の不飽和度制御機構の解明
林祥平, 佐藤康治, 小笠原泰志, 大利徹, 日本農芸化学会東北支部大会プログラム・講演要旨集, 153rd, 71, 22 Sep. 2018
Japanese

多価不飽和脂肪酸合成酵素の機能改変
池内健心, 林祥平, 大塚慎, 小林洸太, 佐藤康治, 大利徹, 日本農芸化学会東北支部大会プログラム・講演要旨集, 153rd, 71, 22 Sep. 2018
Japanese

抗酸化物質エルゴチオネインの発酵生産
佐藤康治, 河野祐介, 大津厳生, 大利徹, バイオサイエンスとインダストリー, 76, 5, 402‐403, 10 Sep. 2018
Japanese

Fermentative overproduction of ergothioneine, a sulfur-containing antioxidant by metabolic engineering of cysteine-overproducing Escherichia coli
大津厳生, 河野祐介, 佐藤康治, 大利徹, Fragrance Journal, 46, 7, 65‐70, 15 Jul. 2018
Japanese

異種発現によるバナナ病原菌由来ジテルペン類の生合成研究
田澤聡大, YE Ying, 尾崎太郎, LIU Chengwei, 南篤志, 小笠原泰志, 大利徹, 及川英秋, 日本農芸化学会大会講演要旨集(Web), 2018, 2018

Studies on the Biosynthesis and Heterologous Production of Fungal Terpenoids
尾崎太郎, 山根桃華, 田澤聡大, YE Ying, LIU Chengwei, 小笠原泰志, 大利徹, 南篤志, 及川英秋, 天然有機化合物討論会講演要旨集(Web), 60th, 2018

カルボニルメチレン構造を有するシュードトリペプチド(Ketomemicin)の生合成機構
大利徹, 日本農芸化学会大会講演要旨集(Web), 2018, 2018

バナナ病原菌が感染時に生産するジテルペン類の異種生産
田澤聡大, YE Ying, 尾崎太郎, LIU Chengwei, 南篤志, 小笠原泰志, 大利徹, 及川英秋, 日本化学会春季年会講演予稿集(CD-ROM), 98th, 2018

ペプチドグリカンの新しい生成機構
大利徹, 日本農芸化学会大会講演要旨集(Web), 2018, 2018

放線菌が生み出した疑似ペプチド化合物-疑似ペプチド（ケトメミシン）がもつカルボニルメチレンの生合成を解明-　　　　　　　　　　　　　　　
小笠原 泰志, 大利 徹, 化学と生物, 56, 76, 78, 2018
Japanese, Introduction scientific journal

多価不飽和脂肪酸合成酵素における3‐ケトアシルリダクターゼとデヒドラターゼの機能解析
林祥平, 佐藤康治, 小笠原泰志, 氏原哲朗, 大利徹, 日本生物工学会大会講演要旨集, 69th, 322, 08 Aug. 2017
Japanese

大腸菌によるErgothioneineの発酵生産
大澤怜, 上出倫敬, 佐藤康治, 河野祐介, 大津厳生, 大利徹, 日本生物工学会大会講演要旨集, 69th, 323, 08 Aug. 2017
Japanese

細菌ペプチドグリカンの新規生合成機構　　　　　　　　　　　　　　　
佐藤康治, 馮 若茵, 大利 徹, B & I, 75, 422, 423, 2017
Japanese, Introduction scientific journal

創薬を志向した微生物の生合成工学　　　　　　　　　　　　　　　
大利 徹, The Japanese Journal of Antibiotics, 71, 1, 1, 11, 2017
Japanese, Introduction scientific journal

ペプチド結合を触媒する新たな酵素　　　　　　　　　　　　　　　
小笠原 泰志, 大利 徹, ペプチド医薬品のスクリーニング・安定化・製剤化技術 (技術情報協会), 2017
Japanese, Introduction commerce magazine

Bioengineering utilizing microorganisms
大利 徹, 生化学, 89, 2, 221, 229, 2017
日本生化学会, Japanese, Introduction scientific journal

ペプチド系天然化合物JBIR‐78およびJBIR‐95の生合成研究
竹田薫平, 佐藤康治, 小笠原泰志, 新家一男, 大利徹, 日本放線菌学会大会講演要旨集, 31st, 80, 08 Sep. 2016
Japanese

抗酸化物質Ergothioneineの酵素合成法の開発
大澤怜, 上出倫敬, 佐藤康治, 大津厳生, 大利徹, 日本放線菌学会大会講演要旨集, 31st, 141, 08 Sep. 2016
Japanese

Photobacterium profundum由来エイコサペンタエン酸合成酵素の異種宿主発現
中真以, 林祥平, 佐藤康治, 氏原哲朗, 大利徹, 日本生物工学会大会講演要旨集, 68th, 253, 25 Aug. 2016
Japanese

多価不飽和脂肪酸合成酵素におけるタンデムアシルキャリアプロテインの機能解明
林祥平, 佐藤康治, 氏原哲朗, 大利徹, 日本生物工学会大会講演要旨集, 68th, 253, 25 Aug. 2016
Japanese

多価不飽和脂肪酸合成酵素の機能解析
林祥平, 佐藤康治, 氏原哲朗, 大利徹, 日本生物工学会大会講演要旨集, 68th, 168, 25 Aug. 2016
Japanese

放線菌に見出されたペプチドライゲースオルソログの機能解析
小笠原泰志, 川田純平, 佐藤康治, 降旗一夫, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2016, 4SY05‐3 (WEB ONLY), 05 Mar. 2016
Japanese

ペプチド系天然化合物JBIR‐78および95の生合成研究
竹田薫平, 佐藤康治, 小笠原泰志, 新家一男, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2016, 2B038 (WEB ONLY), 05 Mar. 2016
Japanese

Xanthomonas oryzaeに見出した新規アミノ酸異性化酵素
FENG Ruoyin, 佐藤康治, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2016, 2B040 (WEB ONLY), 05 Mar. 2016
Japanese

放線菌におけるエルゴチオネイン特異的輸送体の探索
上出倫敬, 佐藤康治, 河野祐介, 大津厳生, 大利徹, 日本農芸化学会北海道支部講演会講演要旨, 2016, 2016

New gene responsible for para-aminobenzoate biosynthesis
佐藤 康治, 倉都 将宏, 小林 大毅, 大利 徹, 生物工学会誌 : seibutsu-kogaku kaishi, 94, 2, 72, 72, 2016
日本生物工学会, Japanese

微生物が生産する生理活性物質研究 : 生物工学的観点から (大村智先生ノーベル賞受賞記念特集 微生物由来天然物の実用化と未来)
大利 徹, 生物工学会誌, 94, 7, 390, 392, 2016
日本生物工学会, Japanese

ペプチド系天然化合物JBIR‐95の生合成研究
竹田薫平, 佐藤康治, 小笠原泰志, 新家一男, 大利徹, 日本農芸化学会北海道支部講演会講演要旨, 2015, 12, 14 Nov. 2015
Japanese

多価不飽和脂肪酸生産能に対するacyl carrier proteinドメイン数の影響
林祥平, 佐藤康治, 氏原哲朗, 大利徹, 日本生物工学会大会講演要旨集, 67th, 337, 337, 25 Sep. 2015
日本生物工学会, Japanese

芳香族アミノ酸水酸化酵素の物質生産への応用
佐藤康治, 大利徹, 日本生物工学会大会講演要旨集, 67th, 352, 352, 25 Sep. 2015
日本生物工学会, Japanese

シアノバクテリアにおけるエルゴチオネイン生合成遺伝子の探索
大澤怜, 佐藤康治, 中嶋駿介, 大津厳生, 高木博史, 大利徹, 日本生物工学会大会講演要旨集, 67th, 336, 336, 25 Sep. 2015
日本生物工学会, Japanese

バクテリア由来フェニルアラニン水酸化酵素を利用したチロシン生産
佐藤康治, 大利徹, 日本生物工学会大会講演要旨集, 67th, 337, 337, 25 Sep. 2015
日本生物工学会, Japanese

緩い基質特異性を示すペプチドリガーゼの結晶構造からの考察と応用
NOIKE MOTOYOSHI, MATSUI TAKASHI, OGASAWARA YASUSHI, MORITA HIROYUKI, DAIRI TOORU, 日本農芸化学会大会講演要旨集(Web), 2015, 3E11P02 (WEB ONLY), 05 Mar. 2015
Japanese

C‐末にD‐Trpを持つラッソペプチドの生合成機構
NOMURA SATOSHI, OGASAWARA YASUSHI, SATO YASUHARU, DAIRI TOORU, 日本農芸化学会大会講演要旨集(Web), 2015, 3E11P05 (WEB ONLY), 05 Mar. 2015
Japanese

3S-Ba05 Amido bond formation catalyzed by newly discovered ATP-grasp-ligases
Dairi Tohru, 日本生物工学会大会講演要旨集, 67, 344, 344, 2015
日本生物工学会, Japanese

Studies on the new menaquinone biosynthetic pathway found in microorganisms and application for drug discovery to pathogenic Helicobacter and Chlamydia strains
大利 徹, 旭硝子財団助成研究成果報告 Reports of research assisted by the Asahi Glass Foundation, 1, 4, 2015
旭硝子財団, Japanese

A new enzyme catalyzing amide bond formation
大利 徹, バイオサイエンスとインダストリー, 73, 4, 274, 279, 2015
バイオインダストリー協会, Japanese

放線菌に見いだされた新規ペプチドライゲースは幅広い基質特異性を持つ
NOIKE MOTOYOSHI, MATSUI TAKASHI, SATO YASUHARU, MORITA HIROYUKI, DAIRI TOORU, 日本農芸化学会北海道支部講演会講演要旨, 2014, 30, 22 Sep. 2014
Japanese

インドールジテルペンの生合成に関与するプレニル基転移酵素に関する研究
RYU SEII, FUJIMORI MICHIKO, MINAMI ATSUSHI, NOIKE MOTOYOSHI, SATO YASUHARU, OGURI HIROKI, OIKAWA HIDEAKI, DAIRI TOORU, イソプレノイド研究会例会講演要旨集, 24th, 19, 12 Sep. 2014
Japanese

バクテリアにおける新規葉酸生合成関連遺伝子の同定
SATO YASUHARU, KOBAYASHI DAIKI, DAIRI TOORU, 日本生物工学会大会講演要旨集, 66th, 162, 162, 05 Aug. 2014
日本生物工学会, Japanese

エルゴチオネインは酸化ストレスから放線菌を守る
NAKAJIMA SHUNSUKE, SATO YASUHARU, DAIRI TOORU, 日本生物工学会大会講演要旨集, 66th, 162, 162, 05 Aug. 2014
日本生物工学会, Japanese

放線菌に見いだされた新規ペプチドライゲースは幅広い基質特異性を持つ
NOIKE MOTOYOSHI, MATSUI TAKASHI, SATO YASUHARU, MORITA HIROYUKI, DAIRI TOORU, 日本放線菌学会大会講演要旨集, 29th, 92, 19 Jun. 2014
Japanese

ペプチドライゲースとリボソームとの協同によるペプチド新奇生合成機構の解明
NOIKE MOTOYOSHI, OYA KOICHI, SASAKI IKUO, MARUYAMA CHITOSE, ISHIKAWA JUN, SATO YASUHARU, HAMANO YOSHIMITSU, ITO HAJIME, DAIRI TOORU, 日本農芸化学会大会講演要旨集(Web), 2014, 2A04P08 (WEB ONLY), 05 Mar. 2014
Japanese

インドールジテルペンの生合成に関与するプレニル基転移酵素に関する研究―2
LIU CHENGWEI, FUJIMORI MICHIKO, MINAMI ATSUSHI, NOIKE MOTOYOSHI, SATO YASUHARU, OGURI HIROKI, OIKAWA HIDEAKI, DAIRI TOORU, 日本農芸化学会大会講演要旨集(Web), 2014, 2A04P10 (WEB ONLY), 05 Mar. 2014
Japanese

エルゴチオネインはStreptomyces coelicolorを酸化ストレスから守る
NAKAJIMA SHUNSUKE, SATO YASUHARU, YANASHIMA KENTARO, MATSUI TOMOMI, DAIRI TOORU, 日本農芸化学会大会講演要旨集(Web), 2014, 2A04P11 (WEB ONLY), 05 Mar. 2014
Japanese

2S-Da05 A novel peptide ligase catalyzing a capping of N-terminus of peptides
Noike Motoyoshi, Ooya Koichi, Dairi Tohru, 日本生物工学会大会講演要旨集, 66, 98, 98, 2014
日本生物工学会, Japanese

Peptide synthesis cooperatively achieved by peptide ligase and ribosomes
Noike Motoyoshi, Morita Hiroyuki, Dairi Tohru, Matsui Takashi, Ooya Koichi, Sasaki Ikuo, Maruyama Chitose, Hamano Yoshimitsu, Ishikawa Jun, Satoh Yasuharu, Ito Hajime, Symposium on the Chemistry of Natural Products, symposium papers, 56, 0, Oral8, 2014

Peptide antibiotics are biosynthesized by two distinct machineries: (i) ribosome-based synthesis, and (ii) ribosome-independent synthesis exemplified by nonribosomal peptide synthesis. Pheganomycin (PGM) consists of the nonproteinogenic amino acid (S)-2-(3,5-dihydroxy-4-methoxyphenyl)-2-guanidinoacetic acid (1) at the N-terminus and a proteinogenic core peptide derived from NVKDGPT or NVKDR. The biosynthetic gene cluster responsible for the catalysis was identified in Streptomyces cirratus, containing a gene encoding a precursor peptide, including both core peptides, and several genes plausibly encoding enzymes for 1 biosynthesis and tailoring reactions. We searched for the gene responsible for the peptide bond formation between 1 and the peptides in the cluster, and identified pgm1, which has an ATP-grasp domain. A pgm1-deletion mutant lost PGM productivity, and recombinant PGM1 catalyzed the ATP-dependent peptide bond formation. This is the first example of cooperative peptide synthesis achieved by ribosomes and peptide ligases using a peptide as a nucleophile. PGM1 accepted a variety of peptides as the nucleophile and its flexibility was comprehended by the crystal structure of PGM1 and mutagenesis analyses.

, Symposium on the Chemistry of Natural Products Steering Committee, Japanese

ペプチド系抗生初質フェガノマイシンの生合成に見いだされた新規ペプチドライゲースPGM1の結晶構造解析
MATSUI TAKASHI, OTAKI SHOTA, NOIKE MOTOYOSHI, DAIRI TOORU, MORITA HIROYUKI, 日本結晶学会年会講演要旨集, 2014, 79, 2014
Japanese

ビタミンKの生合成　　　　　　　　　　　　　　　
大利 徹, ビタミンKと疾患, 40, 46, 2014
Japanese, Introduction commerce magazine

ペプチド系抗生物質フェガノマイシンの生合成に関与する新規ペプチドライゲース
野池基義, 雄谷洸一, 佐々木郁雄, 丸山千登勢, 石川淳, 佐藤康治, 濱野吉十, 伊藤肇, 大利徹, 日本農芸化学会北海道支部講演会講演要旨, 2013, 24, 27 Nov. 2013
Japanese

エルゴチオネインはStreptomyces coelicolorを酸化ストレスから守る
中嶋駿介, 佐藤康治, 簗島謙太郎, 松井智未, 大利徹, 日本農芸化学会北海道支部講演会講演要旨, 2013, 18, 27 Nov. 2013
Japanese

放線菌に見出した新規ペプチドライゲース
野池基義, 雄谷洸一, 佐々木郁雄, 丸山千登勢, 石川淳, 佐藤康治, 濱野吉十, 伊藤肇, 大利徹, 日本放線菌学会大会講演要旨集, 28th, 62, 05 Sep. 2013
Japanese

エルゴチオネインはStreptomyces coelicolorを酸化ストレスから守る
中嶋駿介, 佐藤康治, 簗島謙太郎, 松井智未, 大利徹, 日本放線菌学会大会講演要旨集, 28th, 63, 05 Sep. 2013
Japanese

ペプチド系抗生物質,ボトロマイシン生合成機構の解明
佐藤康治, 大門美保, 簗島謙太郎, 尾仲宏康, 石川淳, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2013, WEB ONLY 3C22A12, 05 Mar. 2013
Japanese

バクテリアにおける新規葉酸生合成経路の解明
佐藤康治, 小林大毅, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2013, WEB ONLY 3C22A11, 05 Mar. 2013
Japanese

ペプチド系抗生物質,フェガノマイシン生合成機構の解明
野池基義, 雄谷洸一, 佐藤康治, 丸山千登勢, 濱野吉十, 石川淳, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2013, WEB ONLY 3C22A13, 05 Mar. 2013
Japanese

放線菌が持つメナキノン新規生合成経路の解明と特異的阻害剤の探索
池田駿介, 池田安由美, 佐藤康治, 野池基義, 瀬戸治男, 大利徹, 日本放線菌学会大会講演要旨集, 27th, 104, 06 Sep. 2012
Japanese

熱安定性酵素を用いたポリヒドロキシアルカン酸のin vitro合成
橋本佳季, HAN Xuerong, 佐藤康治, 田島健次, 田口精一, 大利徹, 高分子学会予稿集(CD-ROM), 61, 2, ROMBUNNO.1PB118, 05 Sep. 2012
Japanese

バイオディーゼル燃料副生成物からのバクテリアセルロースの生産
小瀬亮太, 吉田誠, 砂川直輝, 田島健次, 大利徹, セルロース学会年次大会講演要旨集, 19th, 122, 01 Jul. 2012
Japanese

遺伝子組換え酢酸菌によるセルロース誘導体の合成
佐藤智章, 砂川直輝, 小瀬亮太, 田島健次, 大利徹, セルロース学会年次大会講演要旨集, 19th, 61, 01 Jul. 2012
Japanese

遺伝子組換え酢酸菌によるGlcNAc含有バクテリアセルロースの合成
砂川直輝, 今井友也, YAO Min, 小瀬亮太, 田島健次, 大利徹, セルロース学会年次大会講演要旨集, 19th, 15, 16, 01 Jul. 2012
Japanese

酢酸菌のセルロース合成におけるbglxAおよび下流遺伝子の機能
砂川直輝, 松田恵利子, 小瀬亮太, YAO Min, 田島健次, 大利徹, セルロース学会年次大会講演要旨集, 19th, 133, 01 Jul. 2012
Japanese

バクテリアにおける新規葉酸生合成経路の解明
佐藤康治, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2012, WEB ONLY 2A06A03, 05 Mar. 2012
Japanese

フタロシン(FL)経路の解析
池田駿介, 柏崎恭平, 小野裕介, 池田安由美, 佐藤康治, 矢島新, 瀬戸治男, 大利徹, 日本農芸化学会大会講演要旨集(Web), 2012, WEB ONLY 2A06A02, 05 Mar. 2012
Japanese

Enterobacter sp.CJF002株におけるバクテリアセルロース合成の解析
砂川直輝, 細田真梨子, 田島健次, 河野信, YAO Min, 佐藤康治, 大利徹, セルロース学会年次大会講演要旨集, 18th, 137, 01 Jul. 2011
Japanese

海藻由来ポリヒドロキシアルカン酸合成菌の単離・同定
栗城夢実, 田島健次, 佐藤康治, 尾島孝男, 小林孝紀, 大利徹, 高分子学会北海道支部研究発表会講演要旨集, 45th, 47, 01 Feb. 2011
Japanese

皮膚における脂肪細胞の効果—皮膚を構成する他細胞との相互作用
大木菜津美, 金山寿之, 佐藤康治, 田島健次, 大利徹, 棟方正信, 高分子学会北海道支部研究発表会講演要旨集, 45th, 46, 01 Feb. 2011
Japanese

Studies on an alternative menaquinone biosynthetic pathway operating in actinomycetes
大利 徹, 日本放線菌学会誌, 25, 2, 2, 6, 2011
日本放線菌学会, Japanese

微生物におけるビタミンＫ生合成の分子機構と生物学的意義　　　　　　　　　　　　　　　
大利 徹, 腎と代謝, 24, 179, 186, 2011
Japanese, Introduction commerce magazine

官能基修飾によるポリヒドロキシアルカン酸の機能化
岩本浩介, 谷尾亮, 佐藤康治, 田島健次, 堺井亮介, 佐藤敏文, 鳥谷部哲也, 松島得雄, 覚知豊次, 大利徹, 棟方正信, 棟方正信, 高分子学会予稿集(CD−ROM), 59, 2 Disk1, ROMBUNNO.1PD114, 01 Sep. 2010
Japanese

改良水‐有機溶媒二相反応系による新規ポリヒドロキシアルカン酸の合成
HAN Xuerong, 佐藤康治, 田島健次, 鳥谷部哲也, 松島得雄, 大利徹, 棟方正信, 棟方正信, 高分子学会予稿集(CD−ROM), 59, 2 Disk1, ROMBUNNO.1PC113, 01 Sep. 2010
Japanese

Acetobacter xylinumにおけるbglxA下流遺伝子のクローニングと解析
松田恵利子, 砂川直輝, 吉田誠, 田島健次, 佐藤康治, 棟方正信, 大利徹, セルロース学会年次大会講演要旨集, 17th, 104, 01 Jul. 2010
Japanese

酢酸菌セルロース合成におけるAxCesD N末端の機能解析
砂川直輝, 藤原孝彰, 田島健次, YAO Min, 佐藤康治, 棟方正信, 田中勲, 大利徹, セルロース学会年次大会講演要旨集, 17th, 110, 01 Jul. 2010
Japanese

バイオディーゼル副生成物を原料としたバクテリアセルロース合成
砂川直輝, 吉田誠, 松田恵利子, 細田真梨子, 佐藤康治, 松島得雄, 田島健次, 棟方正信, 大利徹, 日本生物工学会シンポジウム講演要旨集, 2010, 33, 2010
Japanese

33. Characterization of diterpene synthase genes in Aspergillus oryzae
Chiba Yasutaka, Sugai Yoshinori, Shiono Yoshihito, Dairi Tohru, Mitsuhashi Wataru, Kawaide Hiroshi, Toyomasu Tomonobu, 植物化学調節学会研究発表記録集, 44, 47, 47, 06 Oct. 2009
Fungi produce diterpenes exhibiting phytohormone-related activities, such as gibberellin and fusicoccin. We identified a diterpene synthase gene PaFS, which is responsible for fusicoccin biosynthesis, in the mycelium of fusicoccin-producing fungus Phomopsis amygdali (Toyomasu et al., PNAS, 104: 3084-3088, 2007). PaFS (719 amino acids) possesses the diterpene cyclase domain at N-terminus and prenyltransferase domain at C-terminus. Our database search suggested that PaFS homologues exist in genome DNA of several fungal species. Among them, we found two PaFS homologous genes in Aspergillus oryzae genome database. It has been believed that A. oryzae produces few secondary metabolites including terpenoids. In this study, we characterize the two homologues, A. oryzae diterpene synthase like 1(AoDSL1) and AoDSL2. AoDSL1 contained a stop codon in region encoding prenyltransferase domain probably because of naturally occurred nonsense mutation, and is perhaps expressed as a truncated enzyme composed of N-terminal 412 amino acid residues. AoDSL2 encoded 728 amino acid residues, in which a successive Asp of DDxxD motif in the prenyltransferase domain was substituted by AN (position 482-483). These possible naturally occurred mutations perhaps resulted in loss of prenyltransferase activities. Recombinant AoDSL1 converted GGDP into two unknown diterpene cyclic hydrocarbons. The effects of site directed mutagenesis of possible naturally occurred mutation on enzyme activity will be presented., The Japanese Society for Chemical Regulation of Plants, Japanese

微生物に見出されたメナキノン新規生合成経路　　　　　　　　　　　　　　　
大利 徹, 日本生化学会誌, 81, 2, 95, 99, 2009, [Peer-reviewed], [Invited]
Japanese, Introduction scientific journal

細菌に見出されたメナキノン新規生合成経路　　　　　　　　　　　　　　　
大利 徹, バイオサイエンスとインダストリィー, 67, 3, 102, 106, 2009

創薬標的としての新規メナキノン生合成系　　　　　　　　　　　　　　　
大利 徹, 細胞工学, 28, 4, 366, 371, 2009

創薬ターゲットとしてのメナキノン新規生合成経路　　　　　　　　　　　　　　　
大利 徹, ファルマシア, 45, 9, 870, 874, 2009

糸状菌が生産するジテルペン化合物の生合成に関する研究
南篤志, 田島直人, 橋元誠, 加藤雅士, 小林哲夫, 豊増知伸, 佐々武史, 加藤修雄, 大利徹, 日本農芸化学会大会講演要旨集, 2008, 2008

Comparison of enzymatic properties of ent-copalyl diphosphate synthases for biosynthesis of phytoalexins and gibberellins in rice
HAYASHI Yutaka, HIROSE Yuko, ONODERA Yu, MITSUHASHI Wataru, SASSA Takeshi, DAIRI Tohru, TOYOMASU Tomonobu, 植物の生長調節 = Regulation of plant growth & development, 42, 34, 34, 05 Oct. 2007
Gibberellins, a phytohormone, are biosynthesized from geranylgeranyl diphosphate (GGDP) via ent-kaurene. In higher plants, entkaurene was formed from GGDP via ent-copalyl diphsphate (ent-CDP) by two distinct diterpene cyclases, ent-CDP synthase and ent-kaurene synthase. Rice (Oryza sativa L.) produces diterpene phytoalexins, such as oryzalexins, momilactones and phytocassanes, of which biosynthetic hydrocarbon intermediates are converted from GGDP by the similar manner as ent-kaurene synthesis. We have shown that rice has two ent-CDP synthase genes; OsCPSl is for gibberellin biosynthesis and OsCPS2/OsCyc2 is for oryzalexins A-F and phytocassanes A-E. Although ent-CDP synthases for gibberellin biosynthesis from other plant species were well characterized, we have known little about properties of rice ent-CDP synthase for phytoalexin biosynthesis.Here, we enzymatically characterized recombinant putative mature OsCPS2/OsCyc2 as well as OsCPS4/OsCycl, syn-CDP synthase for mimilactones A and B and oryzalexin S, and compared to OsCPS1. The optima of pH (around neutral), temperature (around 30 oC), and divalent ion (0.1 mM Mg^<2+>) on OsCPS2/OsCyc2 and OsCPS4/OsCyc1 were almost the same as OsCPS1. Interestingly, the inhibition rate of activities of OsCPS2/OsCyc2 and OsCPS4/OsCyc1 by substrate, GGDP (50-60μM), and Amo-1618 (10^<-5>-10^<-4>M), a gibberellin biosynthetic inhibitor which suppresses ent-CDP synthase activity, was less than that of OsCPS1., The Japanese Society for Chemical Regulation of Plants, Japanese

リベロマイシンA生合成遺伝子クラスターの解析
高橋俊二, 豊田敦, 鈴木龍一郎, 関山恭代, 植木雅志, 大利徹, 石川淳, 池田治生, 榊佳之, 長田裕之, 日本放線菌学会大会講演要旨集, 22nd, 2007

Nocardia属細菌が持つイソプレノイド生合成遺伝子資源の探索(平成18年度共同利用研究報告)
大利 徹, 三上 襄, 千葉大学真菌医学研究センター報告, 11, 97, 97, 2007
千葉大学, Japanese

2C16-2 An improved procedure for selecting highly active phenylacetaldehyde reductase mutants
MAKINO Yoshihide, DAIRI Tohru, ITOH Nobuya, 日本生物工学会大会講演要旨集, 19, 87, 87, 2007
日本生物工学会, Japanese

Further improvement of engineered phenylacetaldehyde reductase (PAR) on activity under concentrated 2-propanol - Abstracts
Yoshihide Makino, Tohru Dairi, Nobuya Itoh, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 42, 3-4, 116, 117, Nov. 2006
English, Summary international conference

Cloning and expression of cyclo(Leu-Phe) oxidase gene from an actinomycete Streptomyces albulus K023
Machiko Nagao, Atsushi Morimoto, Takashi Tamura, Tohru Dairi, Hiroshi Kanzaki, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 42, 3-4, 126, 126, Nov. 2006
English, Summary international conference

Chiral alcohol production by enantioselective reduction with immobilized recombinant E-coli cells expressing PAR and LSADH - Abstracts
Masatoshi Nakamura, Kousuke Inoue, Yoshihide Makino, Tohru Dairi, Nobuya Itoh, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 42, 3-4, 116, 116, Nov. 2006
English, Summary international conference

Cloning and characterization of a cDNA encoding diterpene cyclase responsible for fusicoccin biosynthesis
TSUKAHARA Mai, TOYOMASU Tomonobu, KANEKO Akane, NIIDA Rie, MITSUHASHI Wataru, DAIRI Tohru, KATO Noboru, SASSA Takeshi, 植物の生長調節 = Regulation of plant growth & development, 41, 61, 61, 04 Oct. 2006
The Japanese Society for Chemical Regulation of Plants, Japanese

Streptomyces sp.CL190株由来の新規Acetoacetyl CoA Thiolaseの機能解析
岡村英治, 大利徹, 葛山智久, 西山真, 日本農芸化学会大会講演要旨集, 2006, 2006

リベロマイシンA生合成遺伝子クラスターの解析
高橋俊二, 関山恭代, 植木雅志, 大利徹, 池田治生, 長田裕之, 日本農芸化学会大会講演要旨集, 2006, 2006

放線菌の多様なイソプレノイド生合成遺伝子　　　　　　　　　　　　　　　
大利 徹, バイオサイエンスとインダストリー, 64, 6, 2006

Studies on biosynthetic genes and enzymes of isoprenoids produced by actinomycetes
DAIRI TOHRU, The Japanese journal of antibiotics, 58, 1, 87, 98, 25 Feb. 2005
日本抗生物質学術協議会, Japanese

リベロマイシンAおよびBの生合成研究:スピロケタールの生成機構について
関山恭代, 植木雅志, 越野広雪, 大利徹, 高橋英俊, 長田裕之, 日本農芸化学会大会講演要旨集, 2004, 2004

Studies on eubacterial diterpene cyclases found in actinomycetes
DAIRI Tohru, HAMANO Toshimitsu, ITOH Nobuya, KUZUYAMA Tomohisa, FURIHATA Kazuo, SETO Haruo, Nippon Nōgeikagaku Kaishi, vol. 76, No. 12, 1191-1194, 12, 1191, 1194, 2002
Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese

テルペノイド抗生物質生産放線菌からのメバロン酸経路遺伝子クラスターのクローニング
濱野 吉十, 大利 徹, 山本 正浩, 川崎 崇, 金田 一秀, 葛山 智久, 伊藤 伸哉, 瀬戸 治男, 日本農芸化学会誌, vol. 76, No. 11, 1092-1093, 11, 1092, 1093, 2002
Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese

Cloning and biochemical characterization of Co2+-activated bromoperoxidase-esterase (perhydrolase) from Pseudomonas putida IF-3 strain
N Itoh, T Kawanami, JQ Liu, T Dairi, M Miyakoshi, C Nitta, Y Kimoto, BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1545, 1-2, 53, 66, Feb. 2001
English

Diversity of microbial threonine aldolases and their application
JQ Liu, T Dairi, N Itoh, M Kataoka, S Shimizu, H Yamada, JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 10, 1-3, 107, 115, Sep. 2000
English, Book review

Studies on antibiotic biosynthetic genes and enzymes catalyzing unique reactions
T Dairi, NIPPON NOGEIKAGAKU KAISHI-JOURNAL OF THE JAPAN SOCIETY FOR BIOSCIENCE BIOTECHNOLOGY AND AGROCHEMISTRY, 74, 9, 975, 983, Sep. 2000
Japanese, Book review

Studies on the nonmevalonate pathway: Conversion of 4-(cytidine 5'- diphospho)-2-C-methyl-D-erythritol to its 2-phospho derivative by 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase
Tomohisa Kuzuyama, Motoki Takagi, Kazuhide Kaneda, Hiroyuki Watanabe, Tohru Dairi, Haruo Seto, Tetrahedron Letters, 41, 16, 2925, 2928, 15 Apr. 2000
English

Studies on the nonmevalonate pathway: formation of 2-C-methyl-D-erythritol 2,4-cyclodiphosphate from 2-phospho-4-(cytidine 5 '-diphospho)-2-C-methyl-D-erythritol
M Takagi, T Kuzuyama, K Kaneda, H Watanabe, T Dairi, H Seto, TETRAHEDRON LETTERS, 41, 18, 3395, 3398, Apr. 2000
English

ユニークな反応を触媒する抗生物質生合成酵素・遺伝子群の解析
大利 徹, 日本農芸化学会誌, 74, 420, 422, 05 Mar. 2000
日本農芸化学会, Japanese

ユニークな反応を触媒する抗生物質生合成酵素・遺伝子の解析
大利 徹, 日本農芸化学会誌, 74, 9, 975, 983, 2000

Formation of 4-(cytidine 5 '-diphospho)-2-C-methyl-D-erythritol from 2-C-methyl-D-erythritol 4-phosphate by 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, a new enzyme in the nonmevalonate pathway
T Kuzuyama, M Takagi, K Kaneda, T Dairi, H Seto, TETRAHEDRON LETTERS, 41, 5, 703, 706, Jan. 2000
English

Alteration of stereoselectivity of L-allo-threonine aldolase by random mutation
Kataoka Michihiko, Kawabata Hiroshi, Liu Ji-Quan, Dairi Tohru, Itoh Nobuya, Yamada Hideaki, Shimizu Sakayu, 日本生物工学会大会講演要旨集, 11, 160, 160, 1999
日本生物工学会, Japanese

Low-specificity L-threonine aldolase of Pseudomonas sp. NCIMB 10558: purification, characterization and its application to beta-hydroxy-alpha-amino acid synthesis
JQ Liu, S Ito, T Dairi, N Itoh, S Shimizu, H Yamada, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 49, 6, 702, 708, Jun. 1998
English

Cofactor characterization of low-specificity D-threonine aldolase
Liu Ji-Quan, Dairi Tohru, Itoh Nobuya, Kataoka Michihiko, Shimizu Sakayu, Yamada Hideaki, 日本生物工学会大会講演要旨集, 10, 168, 168, 1998
日本生物工学会, Japanese

Studies on Antibiotic-Biosynthetic Genes and Enzymes Catalyzing Unique Reatctions.
Tohru Dairi, Actinomycetologica, 12, 2, 75, 83, 1998
日本放線菌学会, English

Protoplasting and regeneration of strains belonging to the genus Actinomadura.
Actinomycetologica, 11, 1, 1, 5, 1997

新しいペニシリン認識酵素、アルカリD-ペプチダーゼの構造と機能 : 酵素
浅野 泰久, 伊藤 元, 大野 泰世, 大利 徹, 加藤 康夫, 日本農藝化學會誌, 70, 301, 301, 05 Mar. 1996
社団法人日本農芸化学会, Japanese

抗真菌抗生物質Pradimicin生合成遺伝子のクローニングと一次構造解析 : 第一報 : 微生物
大利 徹, 古米 保, 沖 俊一, 日本農藝化學會誌, 70, 101, 101, 05 Mar. 1996
社団法人日本農芸化学会, Japanese

Electrophoretic fraction of Ganoderma licidium extract under non-denatured polyacrylamide gel　　　　　　　　　　　　　　　
Journal of Traditional Medicines, 13, 504, 505, 1996

NUCLEOTIDE-SEQUENCE OF FORTIMICIN KL1 METHYLTRANSFERASE GENE ISOLATED FROM MICROMONOSPORA-OLIVASTEROSPORA, AND COMPARISON OF ITS DEDUCED AMINO-ACID-SEQUENCE WITH THOSE OF METHYLTRANSFERASES INVOLVED IN THE BIOSYNTHESIS OF BIALAPHOS AND FOSFOMYCIN
T KUZUYAMA, T SEKI, T DAIRI, T HIDAKA, H SETO, JOURNAL OF ANTIBIOTICS, 48, 10, 1191, 1193, Oct. 1995
English, Report scientific journal

Arthrobacter sp. 1Cのオピン脱水素酵素遺伝子 : 微生物
大利 徹, 浅野 泰久, 日本農藝化學會誌, 69, 270, 270, 05 Jul. 1995
社団法人日本農芸化学会, Japanese

Potential Antifungal and Anti-HIV Agent ; Prodimicin.　　　　　　　　　　　　　　　
┣DBExp. Opin. Patents. (/)-┫DB, 4, 1483, 1491, 1994

Use of a Heterologous Gene for Molecular Breeding of Actinomycetes Producing Structurally Related Antibiotics : Self-defense Genes of Producers of Fortimicin-A(Astromicin)-group Antibiotics.
Toshio Ohata, Tohru Dairi, Eri Hashimoto, Mamoru Hasegawa, ┣DBActinomycetol. (/)-┫DB, 7, 2, 145, 155, 1993

Construction and Application of Gene Cloning System for ┣DBStreptomyces sannanensis. (/)-┫DB　　　　　　　　　　　　　　　
in "Trends in Actinomycetology in Japan", ed. by Yasumasa Koyama, Society for Actinomycetes Japan, Tokyo, 101, 102, 1989

Common Feature beyond the Generic Difference of the Biosynthesis of Fortimicins and Sannamycins.　　　　　　　　　　　　　　　
in "Trends in Actinomycetology in Japan", ed. by Yasumasa Koyama, Society for Actinomycetes Japan, Tokyo, 65, 68, 1989

Cloning and nucleotide sequence of the putative polyketide synthase gene for pradimicin biosynthesis from Actinomadura hibisca
Tohru DAIRI, Yoshimitsu HAMANO, Yasuhiro IGARASHI, Tamotsu FURUMAI, Toshikazu OKI, Biosci. Biotech. Biochem., 61, 9, 1445, 1453, 1977

Isoprenoid Synthesis in Plants and Microorganisms　　　　　　　　　　　　　　　
DAIRI Tohru, Biosynthetic genes and enzymes of isoprenoids produced by actinomycetes
Springer, 2013, [Contributor]

Comprehensive Natural Products II Chemistry and BiologyMander, L., Lui, H.-W., Eds.volume 1, pp. 789–814.　　　　　　　　　　　　　　　
Comprehensive Natural Products II Chemistry and BiologyMander, L., Lui, H.-W., Eds.volume 1, pp. 789–814.
Elsevier: Oxford,, 2010

生合成工学　　　　　　　　　　　　　　　
北海道大学

遺伝子工学特論　　　　　　　　　　　　　　　
富山県立大学

生物工学関連法規　　　　　　　　　　　　　　　
富山県立大学

技術英語　　　　　　　　　　　　　　　
富山県立大学

生物情報学概論　　　　　　　　　　　　　　　
富山県立大学

生物工学概論　　　　　　　　　　　　　　　
富山県立大学

生物工学実験　　　　　　　　　　　　　　　
富山県立大学

分子生物学演習　　　　　　　　　　　　　　　
富山県立大学

分子生物学　　　　　　　　　　　　　　　
富山県立大学

生化学　　　　　　　　　　　　　　　
北海道大学

Dissecting biosynthetic machineries of natural peptides leading to structural and functional diversities.
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S)
27 Apr. 2022 - 31 Mar. 2027
大利 徹, 小笠原 泰志, 森田 洋行, 濱野 吉十
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (S), Hokkaido University, 22H04976

Dissecting reaction mechanisms of novel peptide epimerases
Grants-in-Aid for Scientific Research
01 Apr. 2018 - 31 Mar. 2022
Dairi Tohru
(1) MurL catalyzing the epimerization of the terminus L-Glu of UDP-MurNAc-L-Ala-L-Glu, an intermediate in the alternative biosynthetic pathway of peptidoglycan, was shown to activate the substrate by adenylation of α-carbonyl of the L-Glu. By crystal structure analysis, we were unable to determine the active amino acid residues of MurL, and co-crystal of MurL with a mimic of adenylated substrate was thought to be essential for estimation of the active residues and the reaction mechanism. (2) MslH catalyzed the epimerization of C-terminal L-Trp in a ribosomal peptide MS-271. By crystal structure analysis, active amino acid residues, divalent cation trapping residues, and several substrate recognition residues were identified. (3) Polyglutamate contains D-Glu besides L-Glu. PgsA, a polyglutamate biosynthetic enzyme, was suggested to be the epimerase catalyzing isomerization of terminal L-Glu that is just ligated to the elongating polyglutamate substrate.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 18H03937

Redesigning Biosynthetic Machineries (Project Team)
Grants-in-Aid for Scientific Research
30 Jun. 2016 - 31 Mar. 2021
Abe Ikuro
In this project, we have made a dramatic development from simply learning biosynthetic machineries to designing new blueprints for producing desired natural products. Based on the detailed analyses of the structural diversity of natural products from the viewpoint of genes, enzymes, and reactions, we established a new world-leading technology platform for creation of complex functional molecules by rational reconstruction of biosynthetic machineries as a new academic field. In order to facilitate the research plan, the project team formulated a research policy and provided a forum to actively promote exchanges, collaborations, and joint research among individual research promoters. Researchers from various fields collaborated and complemented each other in their areas of expertise to conduct joint research and create a new academic field.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), The University of Tokyo, 16H06442

Studies on biosynthetic machinery of biologically active natural products
Grants-in-Aid for Scientific Research
30 Jun. 2016 - 31 Mar. 2021
Dairi Tohru
Polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) are essential fatty acids for humans. PUFAs are biosynthesized by either desaturases/elongases from oleic acid or PUFA synthases from acetyl units. PUFA synthases are composed of three or four subunits and each creates a specific PUFA even though the multiple catalytic domains in each subunit are very similar. We dissected these PUFA synthases by in vivo and in vitro experiments and elucidated how the enzymes control PUFA profiles. Moreover, for the first time, we converted a DHA synthase into an EPA synthase by one amino acid substitution.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Hokkaido University, 16H06452

Redesigning Biosynthetic Machineries (International Activity Support Team)
Grants-in-Aid for Scientific Research
30 Jun. 2016 - 31 Mar. 2021
Abe Ikuro
In this project, we have made a dramatic development from simply learning biosynthetic machineries to designing new blueprints for producing desired natural products. We established a new world-leading technology platform for creation of complex functional molecules by rational reconstruction of biosynthetic machineries as a new academic field. In order to strongly promote joint research with foreign countries, the International Activity Support Group invited outstanding foreign researchers and held lecture meetings on a regular basis. Furthermore, international symposiums were held with the aim of strengthening cooperation and exchange with the U.S., Germany, China, and other countries. We strategically promoted joint research with foreign countries, focusing on medium-term support for young researchers. Many of the results of joint research with foreign countries resulted in excellent co-authored papers.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), The University of Tokyo, 16K21725

Biosynthesis of the Carbonylmethylene Structure Found in a Class of Pseudotripeptides, Ketomemicins
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
01 Apr. 2015 - 31 Mar. 2018
Dairi Tohru, SATOH YASUHARU, OGASAWARA YASUSHI, MORITA HIROYUKI
We recently discovered novel pseudotripeptides, ketomemicins, which possess a C-terminal pseudodipeptide connected with a carbonyl methylene instead of an amide bond. The carbonyl methylene structure is stable than amide bond and its biological significance has been shown in several natural and synthetic compounds. Despite the biological significances of these compounds, little is known about its biosynthetic machinery. We therefore examined it by in vitro studies with recombinant enzymes. Consequently, an aldolase, dehydratase, PLP-dependent glycine-C-acetyltransferase, and dehydrogenase were revealed to be involved in the formation of the pseudodipeptide with malonyl-CoA and phenylpyruvate as starter substrates.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 15H03110

Biosynthetic machinery: Deciphering and regulating the system for creating structural diversity of baioactive metabolites
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
01 Apr. 2010 - 31 Mar. 2016
OIKAWA Hideaki, EGUCHI Tadashi, ABE Ikuro, KUZUYAMA Tomohisa, DAIRI Tohru, IKEDA Haruo, GOMI Katsuya, SAITO Kazuki, KANAYA Shigehiko, ISHIKAWA Jun
新学術領域領域代表者と8名の計画研究代表者(江口、大利、阿部、池田、五味、斉藤、金谷、石川)、5名の評価委員の意見を取りまとめ、新学術領域研究「生合成マシナリー」の成果報告シンポジウムを、聴衆が期待できる週末のアクセスが良い場所という観点から、東京都港区のコクヨホールに決定した。6月21日の日曜日には、100名近い企業からの出席者や一般参加者に対し、5年間の研究成果を、A01,A02,A03班の班長および指名された顕著な成果を挙げた合計７名の講演者が、判りやすく説明した。


最終の文部科学省へ提出する成果報告書は、３班構成の各班長が、班員個人より事務局に提出された5年間の領域活動で得られた成果に関する資料をもとにデータを整理し、論文発表、学会発表、シンポジウムや講演会での依頼講演、新聞・テレビなどのメディアでの公表状況、各賞の受賞状況、さらには領域内での共同研究など各班の成果を取りまとめた後、事後評価ヒアリング、さらには一般公開用資料としてデータファイルを作成した。これをベースに冊子体も作成し、公的な研究機関や関連研究者に送付した。


6月から10月オープンキャンパスや夏休み期間中には、領域代表が所属する北海道大学や班員が所属する研究機関で、中高生を対象にオープンラボなどの企画で、研究活動を疑似体験してもらうために、実験を撮影したビデオの上映、さらに実験室の見学を行なった。さらに11月14－15日（土・日）、東京都江東区にある日本科学未来館で開かれた大規模なサイエンスコミュニケーションのイベント、サイエンスアゴラに出展し、総括班でアウトリーチ用に作成した領域活動の広報用アニメーションやクイズ形式の説明用プレゼンファイルを利用して、来場者に研究成果を説明した。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Hokkaido University, 22108001

Peptide synthesis cooperatively achieved by peptide ligase and ribosomes
Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
01 Apr. 2013 - 31 Mar. 2015
DAIRI Tohru
Pheganomycin (PGM) (1) consists of a nonproteinogenic amino acid, (S)-2-(3,5-dihydroxy-4-methoxyphenyl)-2-guanidinoacetic acid (2) at the N-terminus and a proteinogenic core peptide derived from NVKDGPT or NVKDR. The biosynthetic gene cluster was identified in Streptomyces cirratus to contain a gene encoding a precursor peptide, which included both the core peptides, and several genes plausibly encoding enzymes for 2 biosynthesis. We identified a gene (pgm1) responsible for the peptide bond formation between 2 and the peptides in the cluster. A pgm1-disruptant lost 1 productivity and recombinant PGM1 catalyzed the ATP-dependent peptide bond formation. This is the first example of cooperative peptide synthesis achieved by ribosome and peptide ligase using a peptide as a nucleophile. PGM1 accepted a variety of peptides as the nucleophile and the flexibility was comprehended by the crystal structure of PGM1 and the mutagenesis analyses.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 25560397

Studies on biosynthetic machineries for cyclic terpenoids and nucleoside antibiotics
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
01 Apr. 2011 - 31 Mar. 2015
DAIRI Tohru, KATO Nobuo, SATOH Yasuharu, ARAI Ryouichi
1. Fusicoccin A is a diterpene glucoside produced by the fungus Phomopsis amygdali and its derivative lacking the OH group at the 12-position is revealed to be a potential anti-cancer drug. We therefore constructed a mutant producing the desirable intermediate by biosynthetic engineering. 2. Three fungal prenyltransferases, PaxC, PaxD, and AtmD, all of which are responsible for biosynthesis of indole diterpene compounds, were characterized. 3. Aminodeoxyfutalosine is the first intermediate in the new menaquinone biosynthetic pathway. We demonstrated that a radical S-adenosyl methionine enzyme (MqnE) catalyzed the addition of the adenosyl radical to the double bond of 3-[ (1-carboxyvinyl)oxy]benzoic acid derived from chorismate by MqnA. 4. During the biosynthetic studies of pheganomycin, we identified a new ATP-grasp enzyme, which phosphorylated the non-proteogenic amino acids with ATP and the successive nucleophilic attack of the peptides.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Hokkaido University, 23108101

Study on a novel folate biosynthesis pathway on micoorganism
Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)
01 Apr. 2012 - 31 Mar. 2014
SATOH Yasuharu, DAIRI Tohru, KURATSU Masahiro, OBAYASHI Daiki
Folate is an essential cofactor in all living cells. para-Aminobenzoate (pABA), a building block of folate, is usually derived from chorismate in the shikimate pathway by reactions of PabABC. We previously suggested that an alternative pathway for pABA biosynthesis would operate in some microorganisms such as Lactobacillus fermentum and Nitrosomonas europaea since these bacteria showed a prototrophic phenotype to pABA despite the fact that there are no orthologs of pabABC in their genome databases. In this study, a gene of unknown function, NE1434, was obtained from N. europaea by shotgun cloning using a pABA-auxotrophic Escherichia coli mutant (dpabABC) as a host. A tracer experiment using [U-13C6]glucose suggested that pABA was de novo synthesized in the transformant. An E. coli dpabABCdaroB mutant carrying the NE1434 gene exhibited a prototrophic phenotype to pABA, suggesting that compounds in the shikimate pathway including chorismate were not utilized as substrates by NE1434.
Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (B), Hokkaido University, 24710239

Estimation of an alternative biosynthetic pathway for co-factors by close investigation of genome databases.
Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
2012 - 2012
DAIRI Tohru, SATOH Yasuharu
The invovment of new enzymes for co-factors biosynthesis was examined. pABA; A function unknown gene, NE1434, was obtained from Nitrosomonas europaea by shotgun cloning experiment using pABA-auxotrophic E. coli mutant (△pabABC) as a host. Glutathione; Streptomyces strains do not use glutathione. However, SCO0910 was confirmed to be glutamate-cysteine ligase and was essential for ergothioneine biosynthesis. Taurine; SCO3035 in S. coelicolor A3(2) was reported to be the first enzyme in taurine biosynthesis (J. Bacteriol. 188, 5561, 2006). To investigate taurine biosynthesis in S. coelicolor A3(2), SCO3416, 2782, and 2017, putative enzymes catalyzing thesecond reaction, were used for in vitro assay. However, all enzymes did not show the expected activity.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 24651235

Screening of natural products which inhibits a new menaquinone biosynthetic pathway
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
2010 - 2012
DAIRI Tohru
The futalosine pathway is operating in some bacteria for the biosynthesis of menaquinone. Futalosine is converted into dehypoxanthinyl futalosine (DHFL) by MqnB.In this study, three routes to the formation of DHFL were suggested. DHFL may have been directly formed by MqnB in Thermus thermophilus. In Streptomyces coelicolor, aminodeoxy FL (AFL) was converted to FL by deaminases, then to DHFL. In contrast, MqnB of Helicobacter pylori directly converted AFL into DHFL.
To identify compounds that specifically inhibit the futalosin pathway, we used B. halodurans that has FL pathway as a test strain. Finally, two candidate fungal culture broths were obtained.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 22310139

微生物が生産する環状テルペノイド生合成マシナリーの解明と再構築
科学研究費助成事業 新学術領域研究(研究領域提案型)
2011 - 2011
大利 徹
日本学術振興会, 新学術領域研究(研究領域提案型), 北海道大学, 23108501

病原菌が持つメナキノン新規生合成経路の全容解明と経路特異的阻害剤の探索
科学研究費助成事業 特定領域研究
2008 - 2009
大利 徹
メナキノン(ビタミンK)は、微生物にとって電子伝達系成分として生育に必須である。筆者は、Helicobacter属(ピロリ菌)、Campylobacter属、Wolinella属などの病原微生物や、放線菌Streptomyces属などの微生物では、今まで知られていた経路とは全く異なるフタロシン経路で生合成されることを見出した。今年度、本経路の2番目の反応を触媒するfutalosine hydrolase(MqnB、最近EC3.2.2.26が付与された)の諸性質を高度好熱菌Thermus thermophilusの組換え酵素を用いて検討した。その結果、以下の性質を示した。(1)Futalosineのみが基質となり他の核酸類縁体は基質にならないこと、(2)4量体を形成、(3)至的pHは4.5、(4)至的温度は80度、(5)Km値154.0±5.3μM、kcat1.02/s、(6)hypoxanthineにより弱く阻害されること(Ki値1.1mM)。本酵素がhypoxanthineにより弱く阻害されたことから、hypoxanthine誘導体を合成し、本酵素特異的阻害剤を探索することにより、抗ピロリ菌剤の開発が可能になると期待された。
また天然物からの抗ピロリ菌リード化合物の探索も行った。大学設備ではピロリ菌を培養できないため、2種類のBacillus属細菌を用いた系で一次スクリーニングを行った。同じBacillus属に属しながら、Bacillus haloduransがメナキノン生合成の際、新規経路を使うのに対し、Bacillus subtilisは既知経路を使う。そこで、(1)B.haloduransに対し抗菌作用を示すが、B.subtilisには影響を及ぼさない化合物を放線菌・カビの培養液中に探索した。次に、(2)B.haloduransに対する生育阻害が、外からメナキノンを添加した際、回復するか検討した。その結果、放線菌の培養中に1つの候補化合物を見出した。現在、本化合物の精製と構造解析を行っている。
日本学術振興会, 特定領域研究, 富山県立大学, 20018023

Studies on diterpene cyclases found in eubacteria
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
2007 - 2009
大利 徹
ジテルペン化合物は(炭素数20)、香料、医薬・医薬中間体、植物ホルモンなど重要な化合物を含んでいる。これら化合物の生合成の第一段階は、環化酵素が直鎖状の基質であるゲラニルゲラニル2 リン酸の末端オレフィンをプロトン化、または2 リン酸を脱離することによってカルボカチオンを生成することから始まる。最終的にカルボカチオンが捕捉中性化されるまで、各環化酵素特有のカチオン中間体を経る反応が順次進行し、多種多様なイソプレノイド骨格へと導かれる。反応の第一段階が共通であるにもかかわらず、最終的に多様な骨格を持つ化合物が生成する事実は、環化酵素の特定アミノ酸残基のみが反応に関与するのではなく、酵素全体のアミノ酸残基が種々のカルボカチオン中間体の生成に関与することを示唆する。従って、これら環化酵素の構造機能相関解析を行うことにより、任意の段階でカルボカチオンが捕捉中性化された化合物の生成を制御できる可能性がある。しかし環化酵素に関しては、真核生物を材料に用いた場合、酵素調製が難しいことなどから一部の酵素を除いて殆ど解析が行われていない。このような背景下、筆者は原核生物と真核生物起源のイソプレノイド生合成遺伝子を多数取得しており、今回、(1)お米由来の2 つのent-copalyl diphosphate 生合成酵素、(2)原核生物のNocardia 属放線菌が生産するジテルペン化合物、brasilicardin A 生合成遺伝子クラスターの取得、(3)原核生物のStreptomyces 属放線菌により生産されるテトラテルペン化合物(KS-505a)生合成遺伝子クラスターの取得とこれらの機能解析を行った。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Toyama Prefectural University, 19510220

病原菌が持つメナキノン新規生合成経路の全容解明と経路特異的阻害剤の探索
科学研究費助成事業 特定領域研究
2006 - 2007
大利 徹
メナキノン(ビタミンK)は,人間にとっては血液凝固に必要なビタミンであり,また微生物では電子伝達系成分として生育に必須である。申請者は,胃潰瘍・胃がんの原因菌として知られているHelicobacter属細菌、食中毒原因菌として知られているCampylobacter属細菌,グラム陽性の土壌細菌であるStreptomyces属細菌などの微生物では,既知の生合成経路として知られているコリスミ酸からメナキノンに至る5ステップの生合成遺伝子群が全く存在しないことに気づいた。そこで,この新規経路の全容解明を行っている。
1.これまでにStreptomyces属放線菌を材料に用いて,バイオインフォマティクスにより絞り込んだ遺伝子を破壊することにより,7つの遺伝子群が新規経路に関与することを明らかにした(これらの成果については未発表のため,これら遺伝子群を以下men06,men26,men27,men50,men90,men92,men94と略号で記載する)。
2.また新規経路遺伝子の網羅的取得を目指し,変異剤によるメナキノン生合成欠損株の誘導と相補遺伝子の取得も行った結果,新規経路は既知経路同様コリスミ酸を出発基質とするが,その後全く別経路を経ることも分かった。
3.上述の破壊株を2つ組み合わせてメナキノン非存在下で混合培養を行った結果,幾つかの菌株の組み合わせで生育が認められた。本現象を利用し,破壊株が関与する生合成上の相対的な位置を(men06)→(men26,men27,men50)→(men90,men92,men94)→メナキノンのように明らかにすることができた。
4,上述したアッセイ系を用い,破壊株が蓄積する中間体の精製・構造決定を行い,3つの化合物の構造を決定した。
日本学術振興会, 特定領域研究, 富山県立大学, 18018035

Studies on a new biosynthetic pathway for menaquinone (vitamin K)
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
2005 - 2007
SETO Haruo, SUE Masayuki, DAIRI Toru
We found that some pathogenic bacteria including Helicobacter pylori and Campylobacter jejuniutilized a new biosynthetic pathway for menaquinone. This pathway is also present in the genus Streptomyces and branches from the well known menaquinone biosynthetic pathway at chorismate. Since this pathway does not present in human, its inhibitors are expected to be useful as antibacterial substances. In order to reveal the details of this pathway, we carried out the following experiments.
1. At the onset of the experiments, we established unequivocal assignment of 13C-N1VIR spectrum of menaquinone.
2. Labeling experiments using glucoses labeled by^<13>C at different position proved that menaquinone was formed from erythrose, phosphoenolpyruvate and two different C_2 units originating from C5-C6 of glucose.
3. Several kinds of blocked mutants of Streptomyces coelicolor that required menaquinone for their growth were prepared.
4. Based on the results obtained by labeling experiments, 1,4-naphthoquinone-6-carboxylic acid was assumed to be a biosynthetic intermediate of the new pathway. Therefore, we prepared this acid by oxidation of naphthalene-2-carboxylic acid by using Ce(SO_4)_2. This compound enabled the growth of a menaquinone auxotroph of S coelicolor indicating that the synthetic compound is a true intermediate of the new pathway.
5. By detailed investigation of metabolites accumulated by mutants of S coelicolor a nucleoside compound, futalosine, was isolated. This compound consisted of inosine and a m-substituted benzoic acid derivative, and had been isolated from another strain of Streptomyces as an antitumor substance. This compound was proved to be a biosynthetic intermediate of the new pathway, since it supported the growth of some menaquinone auxotrophs of S coelicolor.
6. By utilizing other menaquinone auxotrophs of S coelicolor, several intermediates after futalosine were identified.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Tokyo University of Agriculture, 17380075

Studies on production and improvement of terpenoids using biosynthetic gene cluster in fungi and plants.
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
2004 - 2006
SASSA Takeshi, YAMANE Hisakazu, HOSHINO Tsutomu, KATO Nobuo, OIKAWA Hideaki, DAIRI Tohou
A cDNA encoding fusicoccadiene synthase (PaFS) was isolated from mycelia of fusicoccin-producing fungi. PaFS possesses two domain, terpene cyclase domain at N-termini and prenyltransferase domain at C-termini, and shows multifunctional catalyzation. We have clarified the detailed cyclization mechanism forming fusicoccane skeleton from GGDP, and have succeeded in synthesizing a fucicoccin derivative that has a differentiation inducing activity. Mechanism on construction of molecular skeleton of diterpene, phomaction was investigated by chemical methods. Heterologous expression of the genes responsible for biosynthesis of diterpene, aphidicolin using yeast was explored. A common biosynthetic intermediate of indole diterpenoide was synthesized as non-and deuterium-labeled form. The deuterium-labeled intermediate was converted into paxilline and emindole DA via epoxidation and cyclization in two different fungi in an intact manner. These results give first evidence that various indole diterpenoide are biosynthesized from a common intermediate. We have cloned three diterpene cyclase genes and one tetraterpene cyclase gene from actinomycetes. Of the four cyclases, enzymatic properties of the two diterpene cyclases were investigated with recombinant enzymes. The functional analyses were performed in respect to Rv3377c and Rv3378c involved in Mycobacterium tuberculosis H37 strain. These genes encode diterpene synthases for producing halimane skeleton. Studies on squalene cyclase were also conducted to gain more insights into the catalytic mechanism. Successful site-specific incorporation of fluorophenylalanines into the catalytic sites demonstrated the importance of cation-pi interaction. In vitro translation system revealed the function of a diterpene cyclase gene from the moss Physcomoitrella patens. This system was also available for determination of production and localization of cytochrome P450 monooxygenase proteins. The function of P450 monooxygenases responsible for biosynthesis of plant hormones, brassinosteroid and gibberellin, was revealed by heterologous expression in Pichia pastoris. We identified 6 cDNAs encoding diterpene cyclases responsible for phytoalexin bisynthesis in rice. We indicated strong evidence for the presence on chromosome 4 of a gene cluster involved in momilactone biosynthesis. The presence of phytocassane biosynthesis gene cluster on chromosome 2 was also suggested.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Yamagata University, 16208012

Studies on isoprenoid biosynthetic genes isolated from prokaryotes
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
2004 - 2006
DAIRI Tohru, MATSUURA Nobuyasu
We have cloned a DNA fragment that contained four ORFs and was confirmed to participate in viguiepinol {3-hydroxypimara-9(11),15-diene} biosynthesis by a heterologous expression experiment, from Streptomyces sp.strain KO-3988. Of the four ORFs, ORF2 and ORF4 were confirmed to encode an ent-copalyl diphosphate(CDP) synthase and a geranylgeranyl diphosphate(GGDP) synthase, respectively, by experiments using recombinant enzymes. ORF3,that did not show similarities with any other known proteins was also expressed in E.coli and used for functional analysis. The purified ORF3 product clearly converted ent-CDP into pimara-9(11),15-diene. Since ORF2 and ORF3 are the first examples of enzymes with these biosynthetic functions from prokaryotes, enzymatic properties of both enzymes were investigated. ORF2 is likely to be a dimer and requires a divalent cation such as Mg^<2+> and Zn^<2+> for its activity. The optimum pH and temperature were 5.5 and 35℃. The Km value was calculated to be 13.7±1.0μM for GGDP and the kcat value was 3.3 x 10^<-2>/sec. ORF3 is likely to be a monomer and also requires a divalent cation. The optimum pH and temperature were 7.0 and 30℃. The Km value for ent-CDP was estimated to be 2.6±0.2μM and the kcat value was 1.4 x 10^<-3>/sec.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Toyama Prefectural University, 16310153

Studies on diterpene cydases found in actinomycetes.
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
2002 - 2003
DAIRI Tohru
Fubacterial diterpene cyclase genes have been doned from a diterpenoid-antibiotic, terpentecin producer. Their products, ORF11 and ORF12, were confirmed to be essential for the conversion of geranylgeranyl diphosphate (GGDP) into terpentetriene (TTE) that had the same -basic skeleton as terpentecm. Functional analyses of these two enzymes were also performed by using purified recombinant enzymes. The ORF11 product converted GGDP into a cydized intermediate (terpentedienol diphosphate, TDP), and then it was transformed into TTE by the ORF12 product. Interestingly, the ORF12 product directly reacted with GGDP and converted GGDP into three olefinic compounds. Moreover, the ORF12 product reacted even with farnesyl diphosphate (FDP) giving three olefinic compounds, which had the same structures as those formed from GGDP except for the chain-lengths. These results suggested that the ORF11 product with a DXDD motif converted GGDP into TDP by a protonation-initiated cyclization and that the ORF12 product with a DDXXD motif completed the reaction by an ionization-initiated reaction of substrates to an allylic carbocation followed by deprotonation to the olefin. Kinetics of the ORF12 product indicated that the affinity for TDP and GGDP were higher than that of FDP and that the relative activity of the reaction converting TDP into TTE. was highest among the reactions using TDP, GGDP, or FDP as the substrate. These results suggested that an actual reaction catalyzed by the ORF12 was the conversion of TDP into TTE in vivo.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Toyama Prefectural University, 14560074

Scientific research on elucidation of terpene-cyclic reactions in the biosynthesis of bioactive microbial terpenoids.
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
2001 - 2003
SASSA Takeshi, OIKAWA Hideaki, KATO Nobuo, HOSHINO Tsutomu, TOYOMASU Tomonobu, DAIRI Tohru
(1)We isolated (+)-cyatha-3,12-diene from an erinacine-producing fungus, and confirmed its stereostructure by semi-synthesis from erinacine P. The cyathane hydrocarbon could be obtained from GGDP by the cell-free system. From a fusicoccin-mass-producing fungus we isolated (+)-δ-araneosene as the presumed bicyclic hydrocarbon intermediate. (+)-Fusicocca-3(16),10(14)-diene was also isolated and its identification suggested the presence of a new fusicoccenyl cation in the biosynthetic pathway. (2)We identified two gene clusters responsible for diterpene biosynthesis by chromosome waling from GGDP synthase genes from Phomopsis amygdali. The gene clusters led us to successful isolation and characterization of two cDNAs encoding diterpene cyclases. (3)The gene cluster for aphidicolin biosynthesis has been identified by PCR-based genome walking using reported sequence of cDNA coding the diterpene cyclase. Based on non-enzymatic cyclization under various conditions and molecular orbital calculations, cyclization mechanism for construction of aphidicolane skeleton was proposed. (4)We have cloned two diterpene cyclase genes essential for biosynthesis of terpentecin. By using recombinant enzymes, it was revealed that one enzyme converted GGDP into a cyclized intermediate (terpentedienol diphosphate), and then it was transformed into terpentetriene by the other enzyme. (5)By using the site-specific mutants and the substrate analogs, we clearly demonstrated that the steric bulk size of active site residues has an important role in determining the folding conformation of squalene substrate during the polycyclization cascade.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Yamagata University, 13306009

Studies on the biosynthesis of terpenoids produced by streptomycetes.
Grants-in-Aid for Scientific Research
2000 - 2001
DAIRI Tohru
Two gene clusters containing the mevalonate pathway genes and the terpentecin (TP) biosynthetic genes were cloned from Streptomyces griseolosporeus strain MF730-N6, a diterpenoid antibiotic, TP producer. In the former cluster, seven genes enecoding geranylgerairyl diphosphate synthase (GGDPS), mevalonate kinase (MK), mevalonate diphosphate decarbosylase (MDPD), phosphomevalonate kinase (PMK), isopentenyl diphosphate (IPP) isomerase, HMG-CoA reductase, and HMG-CoA synthase were suggested to exist in that order. Heterologous expression of these genes in Ε. coli and Streptomyces Iividans, both of which have only the nonmevalonate pathways, suggested that the genes for the mevalonate paftway were involved in the cloned DNA fragment. The GGDPS, MK, MDPD, PMK, IPP isomerase, and HMG-CoA synthase were expressed in Ε. coli. Among them, the recombinant GGDPS, MK, and IPP isomerase were confirmed to have the expected activities. In the latter cluster, two ORFs, ORF11 and ORF12 that encode proteins showing similarities to eucaryotic diterpene cyclases (DCs) and a eubacterial pentalenene synthase, respectively, were found. The two cyclase genes were expressed in Streptomyces Iividans. The transformant produced a novel cyclic diterpenoid, ent-clerod-3,13(16),14-triene (terpentetriene), which has a common basic skeleton as does TP. The two enzymes were overproduced in Ε. coli and purified to homogeneity. The recombinant ORF11 product converted GGDP into an intermediate with diphosphate, and then it was transformed into terpenteteiene by the recombinant ORF12 product. To the best of our knowledge, this is the first report about a eubacterial DC.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Toyama Prefectural University, 12660083

Cloning and Analysis of Tautomycin Biosynthetic Genes
Grants-in-Aid for Scientific Research
1996 - 1998
UBUKATA Makoto, MATSUURA Nobuyasu, DAIRI Tohru
Tautomycin (TM) isolated from a culture broth of Streptomyces spiroverticillatus is an inhibitor of protein phosphatases 1 and 2A. Since TM and its analogues are expected to be useful probes for elucidating the signal trasduction of mammalian cells, we studied on the absolute structure, conformational analysis, structure-activity relationship, and biosynthesis of the polyketide compound, TM. In the present study, we have studied on the cloning and analysis of TM biosynthetic genes. Here I report the results of experiments done in 1996, 1997 and 1998.
1. Development of transformation system: Protoplast formation and regeneration of the TM producer were achieved by using the reported procedure for other Streptomyces sp. In addition to above experiments, the transformation system was established by curing the producer of the cryptic plasmid by a heat shock treatment. The satisfactory efficiency of 2 x 10ィイD15ィエD1 transformants/μgDNA was obtained using the pIJ702 plasmid, with the result that we optimized various conditions of the transformation system.
2. Cloning of the putative TM biosynthetic gene: The KS and AT primers, which were designed from the published sequences in PKS Type I, were used in PCR strategy to obtain products. The primer design strategy was successful in identifying a fragment from S. spiroverticillatus genomic DNA and a PKS on cosmid pWE15. Sequence analysis of the PCR product strongly suggests that we have cloned a PKS gene flagment in the TM producer.
3. Gene disruption of the putative TM biosynthetic gene: To determine whether the PKS gene fragment obtained from PCR strategy indeed encodes PKS involving in TM biosynthesis, the fragments were then used for gene disruption by recombinational insertion of the cloned 1.8 kb region into the corresponding region on the chromosome. The resulting 15 strains were fermented and assayed for the presence of TM. Although none of the strains produced TM, an another antibiotic xanthostatin was produced by all strains. Genomic Southern hybridization analyses showed that TM biosynthetic genes in these strains were deleted from the genomic DNA.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Toyama Prefectural University, 08456062

Reaction Mechanisms and Application of New Bacterial Enzymes
Grants-in-Aid for Scientific Research
1996 - 1997
ASANO Yasuhisa, KOMEDA Hidenobu, KATO Yasuo, DAIRI Tohru
(1) Phenylalanine dehydrogenase (PheDH) : PheDHs from Bacillus sphaericus, B.badius, Sporosarcina ureae were purified from Escherichia coli transformants in large scales and sent to Prof.Engel (University College Dublin, Ireland) and Prof.Rice (University of Sheffield, U.K.) for X-ray crystallography. Based on informations of X-ray crystallography of glutamate dehydrogenase, chimeric enzymes of PheDH with altered substrate specificities were constructed.
(2) Methylaspartate ammonia-lyase (MAL) : MALs from Enterobacteria such as Enterobacter, Citrobacter, Proteus were purified to homogeneities and their enxymological properties were analyzed in detail. MAL from Citrobacter amalonaticus strain YG-1002 was digested with endo peptidases and N-terminal amino acid sequences were determined. Based on the information, DNA probes were synthesized. The gene for the enzyme was cloned form the genomic library of c.amalonaticus strain YG-1002 by PCR and Southern hybridization. The sequence of the gene was compared with that of a strict anaerobe Clostridium tetanomorphum. MAL was crystallized and sent to Prof.Rice for studies of X-ray crystallography.
(3) Opine dehydrogenase (ODH) : The gene for ODH from Arthrobacter sp.IC was cloned and expressed in E.coli. The enzyme was purified in a large scale and sent to Prof.Rice, and on its X-ray studies were started. The first structure of a (D,L) superfamily member, N-(1-D-Carboxylethyl)-L-norvaline dehydrogenase from Arthrobacter sp.strain 1C,has been solved to 1.8* resolution and the location of the bound coenzyme determined.
(4) Results were presented in some international academic meetings.
Japan Society for the Promotion of Science, Grant-in-Aid for international Scientific Research, Toyama Prefectural University, 08044218

Development of Novel Lyases and Their Application to the Synthesis of Optically Active Compounds
Grants-in-Aid for Scientific Research
1995 - 1996
ASANO Yasuhisa, YAMADA Hideaki, DAIRI Tohru, KATO Yasuo
First, we screened for microorganisms having new hydro-lyases catalyzing hydration of maleic acid and itaconic acid, respectively, to produce D-malic acid and citramalic acid which are food additives and starting materials in synthetic organic chemistry, etc. The bacterial maleate hydratase from Arthrobacter sp. was purified to homogeneity and characterized. The culture and reaction conditions for the production of optically pure D-malate from maleate were studied. Under an optimum condition, 87 grams/liter of D-malate was produced in 20 h. The yield was 72 mole %. (S)-(+)-Citramalate producing activity from itaconate was also studied. A.denitrificans produced (S)-(+)-Citramalate in a 99.9 % enantiomeric excess. Under an optimum condition, 27 g of (S)-(+)-citramalate per liter was produced in 30 h. The yield was 69 mol%.
Second, a screening was carried out for new 3-methylaspartate ammonia-lyases which catalyze the synthesis of L-aspartic acid derivatives from fumaric acid derivatives. We discovered for the first time that 3-methylaspartate ammonia-lyase (MAL) producers are relatively widely distributed in the family of facultatively anaerobic Enterobacteriaceae. Crystalline MALs from Citrobacter freundii, C.amalonaticus, and Morganella morganii, were characterized. Using cell-free extracts of the isolates, optically pure (2S, 3S)-3-methylaspartic, (2S, 3S)-3-ethylaspartic, and (2R, 3S)-3-chloroaspartic acids were synthesized from corresponding fumaric acid derivatives.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Toyama Prefectural University, 07660117

Studies on biosynthesis of terpenoids produced by microorganisms　　　　　　　　　　　　　　　
1996
Competitive research funding

Studies on antibiotic biosynthesis　　　　　　　　　　　　　　　
1994
Competitive research funding

Studies on biosynthesis of natural product　　　　　　　　　　　　　　　
Competitive research funding

Studies on biosynthetic genes and enzymes of isoprenoids produced by microorganisms　　　　　　　　　　　　　　　
Competitive research funding