Characterization of yrpC Gene Product of Bacillus subtilis IFO 3336 as Glutamate Racemase Isozyme
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概要
- 論文の詳細を見る
Glr, the glutamate racemase of Bacillus subtilis (formerly Bacillus natto) IFO 3336 encoded by the gir gene, and YrpC, a protein encoded by the yrpC gene, which is located at a different locus from that of the glr gene in the B. subtilis genome, share a high sequence similarity. The yrpC gene complemented the D-glutamate auxotrophy of Escherichia coli WM335 cells defective in the glutamate racemase gene. Glutamate racemase activity was found in the extracts of E. coli WM335 clone cells harboring a plasmid, pYRPC1,carrying its gene. Thus, the yrpC gene encodes an isozyme of glutamate racemase of B. subtilis IFO 3336. YrpC is mostly found in an inactive inclusion body in E. coli JM109/pYRPC1 cells. YrpC was solubilized readily, but glutamate racemase activity was only slightly restored. We purified YrpC from the extracts of E. coli JM109/pYRPC2 cells using a Glutathione S-transferase Gene Fusion System to characterize it. YrpC is a monomeric protein and contains no cofactors, like Glr. Enzymological properties of YrpC, such as the substrate specificity and optimum pH, are also similar to those of Glr. The thermostability of YrpC, however, is considerably lower than that of Glr. In addition, YrpC showed higher affinity and lower catalytic efficiency for L-glutamate than Glr. This is the first example showing the occurrence and properties of a glutamate racemase isozyme.
- 社団法人日本農芸化学会の論文
- 1999-05-23
著者
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Meguro H
Tohoku Fukushi University
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Soda K
Kansai Univ. Osaka Jpn
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Soda Kenji
Department Of Biotechnology Kansai University
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Soda Kenji
Institute For Chemical Research Kyoto University
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Soda Kenji
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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Soda Kenji
Department Of Biotechnology Faculty Of Engineering Kansai University
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Soda Kenji
Institute For Chemical Resarch Kyoto University
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Misono H
Kochi Univ. Kochi Jpn
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Misono Haruo
Department Of Bioresources Science Faculty Of Agriculture Kochi University
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Misono Haruo
Department Of Bioresource Science Kochi University
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Misono Haruo
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kochi University
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ASHIUCHI Makoto
Research Institute of Molecular Genetics, Kochi University
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Ashiuchi Makoto
Department Of Bioresource Science Kochi University
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Ashiuchi Makoto
Research Institute Of Molecular Genetics Kochi University
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SODA Kenji
Faculty of Engineering, Kansai University
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Soda K
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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Soda K
Department Of Biotechnology Kansai University
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MISONO Haruo
Research Institute of Molecular Genetics, Kochi University
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Sato Kumi
Department Of Bioresources Chemistry Graduate School Of Natural Science And Technology Okayama Unive
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