| 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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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥18,180,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥2,880,000)
Fiscal Year 2009: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2008: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2007: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
Fiscal Year 2006: ¥5,700,000 (Direct Cost: ¥5,700,000)
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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 / Arthrobacter / 6-アミノカプロン酸 / 6-Aminohexanoate oligomer / ナイロンオリゴマー分解酵素 / アミド合成 / 耐熱性酵素 / N-terminal nucleophile hydrolase / maloamidase / 6-Aminohexanoate-cyclic dimer / Agromyces / Kokuria / 6-aminohexanoate-oligomer hydrolase / Ntn amidohydrolase |
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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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