2012 Fiscal Year Final Research Report
Characterization of N-acetyltransferase that can catalyze enantio-selectively acetylation of chiral amines.
| Project/Area Number |
22580083
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | Kobe University |
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Principal Investigator |
TAKENAKA Shinji 神戸大学, 大学院・農学研究科, 准教授 (40314512)
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| Project Period (FY) |
2010 – 2012
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| Keywords | 微生物酵素 |
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| Research Abstract |
The demand for optically active D-2-phenylglycine used to synthesize semisynthetic antibiotics and pesticides has been increasing. Here we report the isolation ofa bacterium of the genus Chryseobacteriumthat selectively transformed the L-form of racemic D,L-2-phenylglycine to (2S)-2-acetylamide-2-phenylacetic acid with a molar yield of 49.9%and an enantiomer excess of >99.5% under optimal culture conditions, consequently resulting in 99.0% pure D-2-phenylglycine remaining in the culture. The enantioselective N-acetylation was catalyzed by an acetyl-CoA-dependent N-acetyltransferase whose synthesis was induced by L-2-phenylglycine. The enzyme differed from previously reported bacterial arylamine N-acetyltransferase in molecular mass and substrate specificity. The relative activity ratio of the enzyme with the substrates L-2-phenylglycine, D-2-phenylglycine, 2-(2-chlorophenyl)glycine, and 5-aminosalicylic acid (a good substrate of arylamine N-acetyltransferase) was 100:0:56.9:5.49, respectively. The biotransformation by the N-acetyltransferase-producing bacterium reported here could constitute a new preparative route for the enzymatic resolution of D,L-2-phenylglycine. We cloned the gene encoding N-acetyltransferase of Chryseobacterium sp. 5-3B which showed high identity with putative N-acetyltransferases of Chryseobacteriumgleum ATCC35910 and Chryseobacteriumsp. CF314.
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