Characterization of d-amino acid aminotransferase from Lactobacillus salivarius

Jyumpei Kobayashi, Yasuhiro Shimizu, Yuta Mutaguchi, Katsumi Doi, Toshihisa Ohshima

研究成果: Contribution to journalArticle査読

18 被引用数 (Scopus)

抄録

We searched a UniProt database of lactic acid bacteria in an effort to identify d-amino acid metabolizing enzymes other than alanine racemase. We found a d-amino acid aminotransferase (d-AAT) homologous gene (UniProt ID: Q1WRM6) in the genome of Lactobacillus salivarius. The gene was then expressed in Escherichia coli, and its product exhibited transaminase activity between d-alanine and α-ketoglutarate. This is the first characterization of a d-AAT from a lactic acid bacterium. L. salivarius d-AAT is a homodimer that uses pyridoxal-5′-phosphate (PLP) as a cofactor; it contains 0.91 molecules of PLP per subunit. Maximum activity was seen at a temperature of 60 °C and a pH of 6.0. However, the enzyme lost no activity when incubated for 30 min at 30 °C and pH 5.5 to 9.5, and retained half its activity when incubated at pH 4.5 or 11.0 under the same conditions. Double reciprocal plots of the initial velocity and d-alanine concentrations in the presence of several fixed concentrations of α-ketoglutarate gave a series of parallel lines, which is consistent with a Ping-Pong mechanism. The Km values for d-alanine and α-ketoglutarate were 1.05 and 3.78 mM, respectively. With this enzyme, d-allo-isoleucine exhibited greater relative activity than d-alanine as the amino donor, while α-ketobutylate, glyoxylate and indole-3-pyruvate were all more preferable amino acceptors than α-ketoglutarate. The substrate specificity of L. salivarius d-AAT thus differs greatly from those of the other d-AATs so far reported.

本文言語英語
ページ(範囲)15-22
ページ数8
ジャーナルJournal of Molecular Catalysis B: Enzymatic
94
DOI
出版ステータス出版済み - 2013

All Science Journal Classification (ASJC) codes

  • 触媒
  • バイオエンジニアリング
  • 生化学
  • プロセス化学およびプロセス工学

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