氧化亚铁硫杆菌中3-异丙基苹果酸脱氢酶的过量生产及底物特异性

Overproduction and substrate specificity of 3-isopropylmalate dehydrogenase from Thiobacillus ferrooxidans.

作者信息

Matsunami H, Kawaguchi H, Inagaki K, Eguchi T, Kakinuma K, Tanaka H

机构信息

Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Japan.

出版信息

Biosci Biotechnol Biochem. 1998 Feb;62(2):372-3. doi: 10.1271/bbb.62.372.

DOI:10.1271/bbb.62.372

PMID:9532798

Abstract

We constructed an overexpression system in Escherichia coli of the leuB gene coding for 3-isopropylmalate dehydrogenase in Thiobacillus ferrooxidans. E. coli harboring the plasmid we constructed, pKK leuB1, produced 17-fold the enzyme protein of the expression system previously used for purification. The substrate specificity of the enzyme was analyzed with synthetic (2R, 3S)-3-alkylmalates. The 3-isopropylmalate dehydrogenase of Thiobacillus ferrooxidans had broad specificity toward the alkylmalates.

摘要

我们构建了一个在大肠杆菌中的过表达系统，该系统用于表达编码氧化亚铁硫杆菌中3-异丙基苹果酸脱氢酶的leuB基因。携带我们构建的质粒pKK leuB1的大肠杆菌产生的酶蛋白是先前用于纯化的表达系统的17倍。用合成的(2R, 3S)-3-烷基苹果酸分析了该酶的底物特异性。氧化亚铁硫杆菌的3-异丙基苹果酸脱氢酶对烷基苹果酸具有广泛的特异性。