Glutamate decarboxylase from Lactobacillus brevis:: Activation by ammonium sulfate

被引：67

作者：

Hiraga, Kazumi ^{[1
]}

Ueno, Yoshie ^{[1
,2
]}

Oda, Kohei ^{[1
]}

机构：

[1] Kyoto Inst Technol, Grad Sch Sci & Technol, Dept Appl Biol, Sakyo Ku, Kyoto 6068585, Japan

[2] Kyoto Prefectural Technol Ctr Small & Medium Ente, Shimogyo Ku, Kyoto 6008813, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2008年 / 72卷 / 05期

关键词：

ammonium sulfate; glutamate decarboxylase (GAD); gamma-aminobutyric acid (GABA); Lactobacillus brevis;

D O I：

10.1271/bbb.70782

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this study, the glutamate decarboxylase (GAD) gene from Lactobacillus brevis IFO12005 (Biosci. Biotechnol Biochem., 61, 1168-1171 (1997)), was cloned and expressed. The deduced amino acid sequence showed 99.6% and 53.1% identity with GAD of L. brevis ATCC367 and L. lactis respectively. The His-tagged recombinant GAD showed an optimum pH of 4.5-5.0, and 54kDa on SDS-PAGE. The GAD activity and stability was significantly dependent on the ammonium sulfate concentration, as observed in authentic GAD. Gel filtration showed that the inactive form of the GAD was a dimer. In contrast, the ammonium sulfate-activated form was a tetramer. CD spectral analyses at pH 5.5 revealed that the structures of the tetramer and the dimer were similar. Treatment of the GAD with high concentrations of ammonium sulfate and subsequent dilution with sodium glutamate was essential for tetramer formation and its activation. Thus the biochemical properties of the GAD from L. brevis IFO12W were significantly different from those from other sources.

引用

页码：1299 / 1306

页数：8

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