2009 Fiscal Year Final Research Report
Structural analysis of nylon oligomer hydrolase and application to enzymatic synthesis of unnatural amides
| Project/Area Number |
18360400
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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 |
Biofunction/Bioprocess
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| Research Institution | University of Hyogo |
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Principal Investigator |
NEGORO Seiji University of Hyogo, 大学院・工学研究科, 教授 (90156159)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEO Masahiro 兵庫県立大学, 大学院・工学研究科, 准教授 (40236443)
KATO Dai-ichiro 兵庫県立大学, 大学院・工学研究科, 助教 (60423901)
HIGUCHI Yoshiki 兵庫県立大学, 大学院・生命理学研究科, 教授 (90183574)
SHIBATA Naoki 兵庫県立大学, 大学院・生命理学研究科, 准教授 (30295753)
NAKANO Hidehiko 兵庫県立大学, 大学院・工学研究科, 教授 (20047608)
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| Project Period (FY) |
2006 – 2009
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| Keywords | ナイロンオリゴマー / X線結晶構造解析 / 酵素進化 / βラクタマーゼ / カルボン酸エステル分解酵素 |
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| Research Abstract |
Arthrobacter sp.strain KI72 produces NylB' carboxylesterase, which is 88% homologous to functional nylon oligomer hydrolase (NylB). Both NylB and NylB' utilize Ser112-Lys115-Tyr215 residues as catalytic triads. NylB' possesses approximately 0.5% of the level of the 6-aminohexanoate linear dimer (Ald)-hydrolytic activity of NylB. Three substitutions (G181D, H266N and D370Y) in NylB'-type carboxylesterase were sufficient to increase the level of the Ald-hydrolytic activity to that of the wild-type NylB enzyme. The triple mutant efficiently catalyzed the amide-synthetic reaction in organic solvent containing low concentration of water by reverse reaction of hydrolysis. We analyzed the structural requirements of the enzymes for effective amide-synthetic reaction.
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Research Products
(12 results)
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[Journal Article] Enzymatic synthesis of nylon-6 units in organic solvents containing low concentrations of water.2010
Author(s)
Kawashima, Y., Yasuhira, K., Shibata, N., Matsuura, Y., Tanaka, Y., Taniguchi, M., Miyoshi, Y., Takeo, M., Kato, D., Higuchi, Y., Negoro, S.
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Journal Title
J. Mol. Cat. B: Enzymatic 64
Pages: 81-88
Peer Reviewed
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[Journal Article] X-ray crystallographic analysis of the 6-aminohexanoate cyclic dimer hydrolase: catalytic mechanism and evolution of an enzyme responsible for nylon-6 byproduct degradation.2010
Author(s)
Yasuhira, K. Shibata, N., Mongami, G., Uedo, Y., Atsumi, Y., Kawashima, Y., Hibino, A, Tanaka, Y., Lee, Y-Ho, Kato, D., Takeo, M., Higuchi, Y., Negoro, S.
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Journal Title
J. Biol. Chem. 285
Pages: 1239-1248
Peer Reviewed
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