2007 Fiscal Year Final Research Report Summary
Discovery of amino acid amide racemase and its use in the dynamic kinetic resolution of amino acid amides
| Project/Area Number |
18380061
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Toyama Prefectural University |
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Principal Investigator |
ASANO Yasuhisa Toyama Prefectural University, Department of Biotechnology, Professor (00222589)
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| Co-Investigator(Kenkyū-buntansha) |
KOMEDA Hidenobu Toyama Prefectural University, Faculty of Engineering, Lecturer (50285160)
YAMANE Takashi Nagoya University, Faculty of Engineering, Professor (80030055)
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| Project Period (FY) |
2006 – 2007
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| Keywords | amino acid amide / racemase / amidase / D-aminooeptidase / aloha-amino-ensilon-canrolactam racemase / D-amino acid |
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| Research Abstract |
1. Amino acid amide racemizing activity was discovered in alpha-amino-epsilon-caprolactam (ACL) racemase from Achromobacter obae.
2. Amino acid amidases acting for various amino mid amides, such as D-aminopeptidase (DAP), D-amino acid amidase (DAA), were isolated from various microbial sources, their primary structures deduced and expression in E. coli were made possible.
3. The enzymatic synthesis of D-alanin from L-alanine amide has been demonstrated by use of D-aminopeptidase (DAP) from Ochrobactrum anthropi C1-38 and ACL racemase. The result of successive enzymatic reaction shows that the combination of ACL racemase and DAP can be applied for dynamic kinetic resolution of DL-amino acid amides to yield D-amino acids.
4. Structure of DAA and directed evolution of ACL racemase: We developed a colorimetric method to measure the activity of ACL racemase with stereospecific amino acid amidase and amino acid oxidase, and 4-aminoantipyrine. The structure of D-amino acid amidase from Ochrobactrum anthropiSV3 was solved at 2.1A resolution.
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[Journal Article] Crystal structures of D-amino acid amidase complexed with L-phenylalanine and with L-phenylalanine amide: Insight into D-stereospecificity of D-amino acid amidase from Ochrobactrum anthropi SV32008
Author(s)
Seiji, Okazaki, Atsuo, Suzuki, Tsunehiro, Mizushima, Hidenobu, Komeda, Yasuhisa, Asano, Takashi, Yamane
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Journal Title
Acta Cryst D64
Pages: 331-334
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Enzymes acting on D-amino acid amides, R. Konno, H.Brueckner, A.D'Aniello, G.Fisher, N. Fujii, and H.Homma (eds.).In DAminoacids:A new frontier2007
Author(s)
Yasuhisa Asano
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Journal Title
amino acid and protein research practicalmethods and protocols-, Hauppauge, New York, U.S.A, Nova Science Publishers
Pages: 579-589
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Crystal structure and function of a D-stereospecific amino acid amidase from Ochrobactrum anthropi SV3, a new member of the penicillin-recognizing proteins2007
Author(s)
Seiji, Okazaki, Atsuo, Suzuki, Hidenobu, Komeda, Shigenori, Yamaguchi, Yasuhisa, Asano, Takashi, Yamane
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Journal Title
J. Mol. Biol 368(1)
Pages: 79-91
Description
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[Book] Enzymes acting on D-amino acid amides, R. Konno, H.Brueckner, A.D'Aniello, G.Fisher, N. Fujii, and H.Homma (eds.).In DAminoacids:A new frontier in amino acid and protein research practicalmethods and protocols-, Hauppauge, New York, U.S.A,2007
Author(s)
Yasuhisa Asano
Total Pages
579-589
Publisher
Nova Science Publishers
Description
「研究成果報告書概要(和文)」より
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