Identification of amino acid residues essential for the catalytic reaction of Bacillus kaustophilus leucine aminopeptidase

被引：7
|
作者：
Chi, MC
Chou, WM
Hsu, WH
Lin, LL [1 ]
机构：
[1] Natl Chiayi Univ, Dept Appl Chem, Chiayi 60083, Taiwan
[2] Natl Chiayi Univ, Grad Inst Biotechnol, Chiayi 60083, Taiwan
[3] Natl Chung Hsing Univ, Inst Mol Biol, Taichung 40227, Taiwan
来源：
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2004年 / 68卷 / 08期
关键词：
Bacillus kaustophilus; leucine aminopeptidase; active-site residues; site-directed mutagenesis;
D O I：
10.1271/bbb.68.1794
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：
The functional significance of amino acid residues Lys-265, Asp-270, Lys-277, Asp-288, Asp-347, Glu-349, and Arg-351 of Bacillus kaustophilus leucine aminopeptidase was explored by site-directed mutagenesis. Variants with an apparent molecular mass of approximately 54 kDa were overexpressed in Escherichia coli and purified to homogeneity by nickel-chelate chromatography. The purified mutant enzymes had no LAP activity, implying that these residues are important for the catalytic reaction of the enzyme.
引用
收藏
页码：1794 / 1797
页数：4
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