Structural characterization of L-amino acid oxidase with high substrate specificity and industrial applications
| Project/Area Number |
24560962
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Okayama University |
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Principal Investigator |
INAGAKI KENJI 岡山大学, その他の研究科, 教授 (80184711)
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| Co-Investigator(Kenkyū-buntansha) |
IMADA Katsumi 大阪大学, 大学院理学研究科, 教授 (40346143)
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| Co-Investigator(Renkei-kenkyūsha) |
TAMURA Takashi 岡山大学, 大学院環境生命科学研究科, 教授 (40253009)
INAGAKI Junko 岡山大学, 大学院医歯(薬)学総合研究科, 助教 (90271056)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | L-アミノ酸オキシダーゼ / L-グルタミン酸オキシダーゼ / L-リシンオキシダーゼ / 基質認識機構 / X線結晶構造解析 / 異種発現系構築 / 基質特異性の改変 / 抗腫瘍性酵素 / L-リシン α-オキシダーゼ / 異種発現系の構築 |
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| Outline of Final Research Achievements |
We determined the crystal structure of two L-amino acid oxidases with high substrate specificity, L-glutamate oxidase (LGOX) from Streptomyces sp. X-119-6 and L-lysine α-oxidase (LysOX) from Trichoderma viride. and we analyzed the molecular mechanism of the substrate recognition of L-amino acid oxidases with high substrate specificity. X-ray crystal structure of LGOX revealed that Arg305 in the active site is the key residue involved in substrate recognition. We created 19 mutant enzymes by substitution of Arg305. Some of them, R305D-LGOX, R305L-LGOX, and R305H-LGOX lost oxidase activity for L-Glu, and exhibited a new oxidase activity for L-Arg, L-His, and L-Tyr, respectively. These progress and findings will provide a great contribution to further industrial application of these enzymes.
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Report
(4 results)
Research Products
(88 results)
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[Journal Article] X-Ray crystallographic evidence for the presence of the cysteine tryptophylquinone cofactor in L-lysine ε-oxidase from Marinomonas mediterranea.2013
Author(s)
Okazaki, S., Nakano, S., Matsui,D., Akaji, S., Inagaki, K., and Asano, Y
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Journal Title
J. Biochem.
Volume: 154(3)
Issue: 3
Pages: 233-236
DOI
Related Report
Peer Reviewed
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[Journal Article] Arg305 of Streptomyces L-glutamate oxidase plays a crucial role for substrate recognition.2012
Author(s)
Utsumi, T., Arima, J., Sakaguchi, C., Tamura, T., Sasaki, C., Kusakabe, H., Sugio S., Inagaki, K.
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Journal Title
Biochem. Biophys. Res. Commun.
Volume: 417
Pages: 951-955
NAID
Related Report
Peer Reviewed
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[Presentation] Pseudomonas putida由来メチオニンγ-リアーゼの結晶構造解析2014
Author(s)
湯之戸俊介, 安田江里, 古谷和大, 福本充樹, 工藤大蔵, 志波智生, 佐藤 暖, 田村 隆, 原田繁春, 稲垣賢二湯之戸俊介, 安田江里, 古谷和大, 福本充樹, 工藤大蔵, 志波智生, 佐藤 暖, 田村 隆, 原田繁春, 稲垣賢二
Organizer
第87回日本生化学会大会
Place of Presentation
京都・国立京都国際会館
Year and Date
2014-10-15 – 2014-10-18
Related Report
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[Presentation] L-リシン酸化酵素の基質複合体構造2014
Author(s)
近藤裕輝, 杉山 成, 川口辰也, 天野万里, 新屋敷健悟, 日下部均, 田村 隆, 稲垣賢二, 今田勝巳
Organizer
第87回日本生化学会大会
Place of Presentation
京都・国立京都国際会館
Year and Date
2014-10-15 – 2014-10-18
Related Report
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[Presentation] Engineering of substrate specifity of L-glutamate oxidase from Streptomyces sp.: directed mutagenesis of Arg305 residue2014
Author(s)
Inagaki, K., Fujino, S., Sano, T., Imada, K., Tamura, T., Kusakabe, H.
Organizer
The Fourth International Conference on Cofactors (ICC-04)
Place of Presentation
University of Parma, Parma, Italy
Year and Date
2014-08-25 – 2014-08-28
Related Report
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[Presentation] Molecular Evolution of Gas Cavity in [NiFeSe]-Hydrogenases Resurrected in silico by Ancestral Sequence Estimation and 3D Modeling by Molecular Dynamics Simulation2014
Author(s)
Tamura, T., Hirano, T., Sato, F., Inagaki, K., Tsunekawa, N.
Organizer
The Fourth International Conference on Cofactors (ICC-04)
Place of Presentation
University of Parma, Parma, Italy
Year and Date
2014-08-25 – 2014-08-28
Related Report
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[Presentation] A novel structure of a cysteine tryptophylquinone-dependent oxidase, L-lysine-ε-oxidase from Marinomonas mediterranea2014
Author(s)
Okazaki, S., Nakano, S., Matsui, D., Akaji, S., Inagaki, K., Asano, S.
Organizer
The Fourth International Conference on Cofactors (ICC-04)
Place of Presentation
University of Parma, Parma, Italy
Year and Date
2014-08-25 – 2014-08-28
Related Report
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[Presentation] Gene cloning, recombinant expression, and purification of L-methionine decarboxylase from Streptomyces sp.5902014
Author(s)
Hayashi, M., Okada, A., Tamura, T., Inagaki, K.
Organizer
The Fourth International Conference on Cofactors (ICC-04)
Place of Presentation
University of Parma, Parma, Italy
Year and Date
2014-08-25 – 2014-08-28
Related Report
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[Presentation] Functional and structural analysis of Peseudomonas putida L-methionine γ-lyase and its C116H Mutant2014
Author(s)
Yasuda, E., Fukumoto, M., Furutani, K., Yunoto, S., Tamura, T., Shiba, T., Sato, D., Harada, S., Inagaki, K.
Organizer
The Fourth International Conference on Cofactors (ICC-04)
Place of Presentation
University of Parma, Parma, Italy
Year and Date
2014-08-25 – 2014-08-28
Related Report
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[Presentation] Recombinant expression, and molecular characterization of L-lysine alpha-oxidase from Trichoderma viride2014
Author(s)
Kenji Inagaki, Marie Amano, Haruka Mizuguchi, Kengo Shinyashiki, Yuki Saitoh, Yoshiteru Hashimoto, Michiko Kobayashi, Hiroki Kondo, Tadahisa Sano, Katsumi Imada, Takashi Tamura, Hitoshi Kusakabe
Organizer
18th INTERNATIONAL SYMPOSIUM FLAVINS AND FLAVOPROTEINS
Place of Presentation
Mahidol University, Phetchaburi, Thailand
Year and Date
2014-07-27 – 2014-08-01
Related Report
Invited
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