Expression and characterization of the carboxyl esterase Rv3487c from Mycobacterium tuberculosis

被引：52

作者：

Zhang, M

Wang, JD

Li, ZF

Xie, J

Yang, YP

Zhong, Y

Wang, HH ^{[1
]}

机构：

[1] Fudan Univ, Sch Life Sci, State Key Lab Genet Engn, Inst Genet, Shanghai 200433, Peoples R China

[2] Fudan Univ, Sch Life Sci, Minist Educ, Key Lab Biodivers Sci & Ecol Engn, Shanghai 200433, Peoples R China

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2005年 / 42卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Mycobacterium tuberculosis; esterase; Rv3487c; LipF; catalytic triad;

D O I：

10.1016/j.pep.2005.03.022

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Rv3487c (lipF), a member of the lipase family of Mycobacterium tuberculosis, is related to virulence of this pathogen. Real-time RT-PCR analysis indicated that Rv3487c was induced at low pH in M. tuberculosis cultured in vitro. The gene of Rv3487c was cloned and expressed as fusion protein in Escherichia coli. After removal of the N-terminal domain of the fusion partner by enterokinase treatment, the effect of pH, temperature, and detergents on the purified enzyme activity and stability was characterized. Rv3487c could efficiently hydrolyze short chain esters. The catalytic triad of Rv3487c consists of residues Ser90, Glu189, and His219 as demonstrated by amino acid sequence alignment, three-dimensional modeling, and site-directed mutagenesis. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：59 / 66

页数：8

共 17 条

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