Purification and Properties of D-Glutamate Oxidase from Candida boidinii 2201
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概要
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An enzyme that oxidizes acidic D-amino acids was purified from the yeast Candida boidinii 2201 to homogeneity as indicated on SDS-polyacrylamide gel electrophoresis. The molecular mass determined by the electrophoresis was 45kDa, while gel filtration of the native enzyme showed a molecular mass of 43 kDa, indicating the native enzyme to be a monomer. The enzyme required FAD for activity and FMN did not replace FAD. The purified preparation had a specific activity of 41.4 μmol/min per mg protein with 20 mM D-glutamate as a substrate. The optimum pH and temperature were 7.0 and 37℃. The K_m for D-glutamate was higher than that for D-aspartate, while the K_<cat>was more markedly higher for the former than for the latter, resulting in a higher k_<cal>/K_m value for D-glutamate. This indicates that D-glutamate is the better substrate and suggests that the enzyme should be named D-glutamate oxidase(EC 1. 4. 3. 7). N-Methyl-D-aspartate was a far poorer substrate than these two compounds. D-Malate was the most potent competitive inhibitor, followed by glutarate and meso-tartrate. The N-terminal sequence of the enzyme was similar to those of FAD enzymes, including D-aspartate and D-amino acid oxidases from various sources.
- 社団法人日本生物工学会の論文
- 1998-06-25
著者
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ODANI Shoji
Department of Biology, Faculty of Science, Niigata University
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Kera Yoshio
Environmental Systems Engineering Nagaoka University Of Technology
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Takahashi Mitsuru
Department Of Food Material Center Laboratory Lotte Co. Ltd
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Taguchi Susumu
Department Of Bioengineering Nagaoka University Of Technology
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FUKUNAGA SADAO
Departments of Bioengineering, Nagaoka University of Technology
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YUNO SHINJI
Departments of Bioengineering, Nagaoka University of Technology
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YAMADA RYO-HEI
Environmental Systems Engineering, Nagaoka University of Technology
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Yamada Ryo-hei
Environmental Systems Engineering Nagaoka University Of Technology
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Odani Shoji
Department Biology Faculty Of Science Niigata University
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Takahashi Mitsuru
Department Of Bioengineering Nagaoka University Of Technology
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Yuno Shinji
Departments Of Bioengineering Nagaoka University Of Technology
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Fukunaga Sadao
Departments Of Bioengineering Nagaoka University Of Technology
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Takahashi Mitsuru
Department of Applied Fine Chemistry, Faculty of Engineering, Osaka University
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Fukunaga Sadao
Department of Chemical Science and Technology, Faculty of Engineering, The University of Tokushima
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