The ywad gene from Bacillus subtilis encodes a double-zinc aminopeptidase

被引：13

作者：

Fundoiano-Hershcovitz, YF

Rabinovitch, L

Shulami, S

Reiland, V

Shoham, G

Shoham, Y ^{[1
]}

机构：

[1] Technion Israel Inst Technol, Dept Food Engn & Biotechnol, IL-32000 Haifa, Israel

[2] Technion Israel Inst Technol, Inst Catalysis Sci & Technol, IL-32000 Haifa, Israel

[3] Hebrew Univ Jerusalem, Dept Inorgan Chem, IL-91904 Jerusalem, Israel

[4] Hebrew Univ Jerusalem, Lab Struct Chem & Biol, IL-91904 Jerusalem, Israel

来源：

FEMS MICROBIOLOGY LETTERS | 2005年 / 243卷 / 01期

基金：

以色列科学基金会;

关键词：

ywad gene; Bacillus subtilis aminopeptidase; catalytic mechanism; Zn-metalloenzyme; substrate specificity;

D O I：

10.1016/j.femsle.2004.12.001

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The yet uncharacterized ywad gene from Bacillus subtilis has been cloned and overexpressed in Escherichia coli. The gene product (BSAP) was purified and shown to be an aminopeptidase. The activity of BSAP was optimal at pH 8.4, the enzyme was stable for 20 min at 80 degreesC and its activity was not affected by serine protease and aspartic protease inhibitors, but was completely diminished by the Zn-chelator 1, 10-phenanthroline. ZnCl2 was able to restore activity, and the binding stochiometry of zinc to apo-BSAP indicated two Zn ions per protein molecule. BSAP exhibited high preference toward p-nitroanilide derived Arg, Lys, and Len synthetic substrates resulting in k(cat)/K-m values of 1-5 x 10(1) s(-1) mM(-1). (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：157 / 163

页数：7

共 34 条

[1] BACILLUS-SUBTILIS AMINOPEPTIDASE - SPECIFICITY TOWARD AMINO-ACID AMIDES, DIPEPTIDES, AND OLIGOPEPTIDES [J].