Isozymic nature of spore coat-associated alanine racemase of Bacillus subtilis

被引:17
作者
Kanda-Nambu, K [1 ]
Yasuda, Y [1 ]
Tochikubo, K [1 ]
机构
[1] Nagoya City Univ, Sch Med, Dept Microbiol, Mizuho Ku, Nagoya, Aichi 4678601, Japan
关键词
amino acid; alanine racemase; isozyme; kinetics; inhibitors; Bacillus subtilis; spore coat;
D O I
10.1007/s007260070076
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
Spore coat-associated alanine racemase of Bacillus subtilis, which converts L-alanine to D-alanine, that is, the germinant to the competitive inhibitor, to regulate spore germination for survival of the organism under unfavorable growth conditions, was examined. The dormant spores, L-alanine-initiated germination of which is inhibited by diphenylamine, were used to characterize the enzyme in the native form because of its unextractablility from dormant spores. The presence of isozymes, Enz-I and Enz-II with Km for L-alanine of about 20mM and 50mM and optimum activity at around 40 degrees C and 65 degrees C, respectively, was proposed. The enzymes were selectively used depending on the L-alanine concentration and the temperature. The pH profiles of the activity (optimun at pH 9.0) and the stability (stable between pH 6-11 at 60 degrees C) were similar, but Enz-II was more heat-stable than Enz-I and the denaturation curve demonstrated a two-domain structure for Enz-II. Sensitivity to D-penicillamine, hydroxylamine and HgCl2 was similar between Enz-I and Enz-II, while that to D-cycloserine, L- and D-aminoethylphosphonic acid, monoiodoacetate and N-ethylmaleimide was different; HgCl2 was the most effective inhibitor among these compounds.
引用
收藏
页码:375 / 387
页数:13
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