Purification and Characterization of an α-Haloketone-resistant Formate Dehydrogenase from Thiobacillus sp. Strain KNK65MA, and Cloning of the Gene(Biochemistry & Molecular Biology)
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概要
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Thiobacillus sp. Strain KNK65MA, which produced an NAD-dependent formate dehydrogenase (FDH) highly resistant to α-haloketones, was newly isolated, i. e., the enzyme showed no loss of activity after a 5-h incubation with α-haloketones, such as ethyl 4-chloro-3-oxobutanoate. The enzyme was also resistant to SH reagents. The enzyme, purified to homogeneity, was a dimer composed of identical subunits. The specific activity was 7.6 u/mg, and the apparent K_m values for formate and NAD^+ were 1.6 and 0.048 mM, respectively. The cloned gene of FDH contained one open reading frame (ORF) of 1206 base pairs, predicted to encode a polypeptide of 401 amino acids, with a calculated molecular weight of 44021 ; this gene was highly expressed in E. coli cells. The deduced amino acid sequence of this FDH had high identity to other bacterial FDHs.
- 社団法人日本農芸化学会の論文
- 2003-10-23
著者
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Hasegawa J
Biochemical Research Laboratories Kanegafuchi Chemical Industry Co. Ltd.
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Hasegawa Junzo
Fine Chemicals Research Laboratories Kaneka Corporation
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NANBA Hirokazu
Fine Chemicals Research Laboratories, Kaneka Corporation
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Hasegawa J
Ichimaru Pharcos Co. Ltd. Gifu Jpn
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Hasegawa Junzo
Life Science Rd Center Kaneka Corporation
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Nanba H
Kaneka Corp. Hyogo Jpn
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Nanba Hirokazu
Fine Chemicals Research Laboratories Kaneka Corporation
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TAKAOKA Yasuko
Fine Chemicals Research Laboratories, Kaneka Corporation
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Takaoka Yasuko
Graduate School Of Biological Sciences Nara Institute Of Science And Technology
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Takaoka Yasuko
Fine Chemicals Research Laboratories Kaneka Corporation
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Hasegawa J
Fine Chemicals Research Laboratories Kaneka Corporation
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Hasegawa Junzo
Life Science R & D Center Kaneka Co.
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Nanba Hirokazu
Fine Chemicals Research Laboratories
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