Characterization of endogenous pyridoxal 5'-phosphate-dependent alanine racemase from Bacillus pseudofirmus OF4(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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An open reading frame of 1100bp in the partially sequenced genome sequence of alkaliphilic Bacillus pseudofirmus OF4 was identified as a putative alanine racemase gene(dadX_<OF4>), which was cloned and expressed in Escherichia coli BL21(DE3). The encoded protein DadX_<OF4> was purified to homogeneity by His_6-tag affinity column, gel filtration and ion-exchange chromatography. The amino acid sequence has highest identity with the known alanine racemase from Oceanobacillus iheyensis HTE831(48%). The protein was a dimeric, endogenous PLP-dependent enzyme, which was demonstrated by absorption spectra and enzyme activity with or without PLP. The racemization temperature optimum was 40℃ and the optimal pH was 10.5. The kinetic parameters K_m and V_<max> at 40℃ of alanine racemase, determined by HPLC analysis, were 41.79mM, 10,500 units/mg for L-alanine and 14.91mM, 3708 units/mg for D-alanine, respectively.
- 社団法人日本生物工学会の論文
著者
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Xue Yanfen
State Key Lab of Microbial Resources, Institute of Microbiology, Chinese Academy of Science
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Ma Yanhe
State Key Lab of Microbial Resources, Institute of Microbiology, Chinese Academy of Science
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OHNISHI Kouhei
Research Institute of Molecular Genetics, Kochi University
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Ohnishi Kouhei
Research Institute Of Molecular Genetics Kochi University
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Ohnishi Kouhei
Res. Inst. Of Molecular Genetics Kochi Univ. Kochi 783-8502 Jpn
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Ma Yanhe
State Key Lab Of Microbial Resources Inst. Of Microbiology Chinese Acad. Of Sci. Beijing 100101 Chn
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Xue Yanfen
State Key Lab Of Microbial Resources Inst. Of Microbiology Chinese Acad. Of Sci. Beijing 100101 Chn
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Ju Jiansong
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences
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Xu Shujing
Life College of Science and Engineering, Shaanxi University of Science and Technology
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Wen Jianxin
College of Life Sciences, Fujian Normal University
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Li Gang
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences
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Wen Jianxin
College Of Life Sciences Fujian Normal University
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Ju Jiansong
Life Coll. Of Sci. And Engineering Shaanxi Univ. Of Sci. And Technol. Xi'an 710021 Chn
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Ju Jiansong
State Key Laboratory Of Microbial Resources Institute Of Microbiology Chinese Academy Of Sciences:li
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Xu Shujing
Life College Of Science And Engineering Shaanxi University Of Science And Technology
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Li Gang
State Key Laboratory Of Microbial Resources Institute Of Microbiology Chinese Academy Of Sciences
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Li Gang
State Key Laboratory Of Coordination Chemistry Department Of Chemistry Nanjing University
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