Analysis of Oxidation Sensitivity of Maleate cis-trans Isomerase from Serratia marcescens
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概要
- 論文の詳細を見る
The maleate cis-trans isomerase gene (maiA) from Serratia marcescens IFO3736 was cloned and sequenced. Serratia MaiA has 62.4% amino acid identity with Alcaligenes faecalis IFO13111 MaiA and 64.9% with Bacillus stearothermophilus MI-102 MaiA. All known ten amino acid sequences of MaiA had significant conserved regions containing cysteine residues, which were previously suggested to be involved in an active site of the enzyme. The maiA gene was expressed in Escherichia coli, and expressed products MaiA was purified and characterized. The purified enzyme of strain IFO3736 showed high activity at room temperature and high heat stability. It also showed higher activity in the presence of high concentration of aspartic acid than the enzyme of A. faecalis IFO13111,but it was also sensitive to chemical oxidation. By amino acid composition analysis, cysteine, methionine, and tyrosine residues were suggested to be oxidized to inactivate the enzyme by chemical oxidation. To investigate the mechanism of chemical oxidation of the enzyme, six methionine residues in the conserved regions of S. marcescens MaiA were replaced with cysteine residues by site-directed mutagenesis. The analysis of the constructed mutants suggested that the Met201 residue near the Cys198 residue is involved in the sensitivity of the enzyme to chemical oxidation.
- 社団法人日本農芸化学会の論文
- 2000-07-23
著者
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Kobayashi Miki
Biochemicals Laboratory Mitsubishi Chemical Corporation
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Goto Machio
Hitachi Instrument Engineering Co. Ltd.
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Goto M
Mie Univ. Tsu Jpn
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HATAKEYAMA Kazuhisa
Biochemicals Laboratory, Tsukuba Research Center, Mitsubishi Chemical Corporation
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GOTO Makoto
Biochemicals Laboratory, Tsukuba Research Center, Mitsubishi Chemical Corporation
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TERASAWA Masato
Biochemicals Laboratory, Tsukuba Research Center, Mitsubishi Chemical Corporation
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YUKAWA Hideaki
Biochemicals Laboratory, Tsukuba Research Center, Mitsubishi Chemical Corporation
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Kobayashi Masatomo
Inst. Phys. Chem. Res. (riken)
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Kobayashi M
Graduate School Of Pharmaceutical Sciences Osaka University
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Kobayashi M
Carbohydrate Science Laboratory National Food Research Institute Ministry Of Agriculture Forestry An
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Terasawa Masato
Biochemicals Laboratory Mitsubishi Chemical Corporation
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Hatakeyama Kazuhisa
Biochemicals Laboratory Mitsubishi Chemical Corporation
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Yukawa H
Res. Inst. Innovative Technol. Earth Kyoto Jpn
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Yukawa Hideaki
Biochemicals Laboratory, Mitsubishi Chemical Corporation
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