来自吡咯菌素假单胞菌的氯过氧化物酶编码基因：序列、在异源宿主中的表达及该酶的纯化

Chloroperoxidase-encoding gene from Pseudomonas pyrrocinia: sequence, expression in heterologous hosts, and purification of the enzyme.

作者信息

Wolfframm C, Lingens F, Mutzel R, van Pée K H

机构信息

Institut f r Mikrobiologie, Universität Hohenheim, Stuttgart, Germany.

出版信息

Gene. 1993 Aug 16;130(1):131-5. doi: 10.1016/0378-1119(93)90356-8.

DOI:10.1016/0378-1119(93)90356-8

PMID:8344520

Abstract

The nucleotide sequence of a 1.5-kb fragment of Pseudomonas pyrrocinia DNA containing the chloroperoxidase(CPO)-encoding gene (cpo) and its flanking regions was determined. The cpo codes for a protein of 278 amino acids (aa). The matuŕe enzyme contains no N-terminal methionine, so that the CPO monomer consists of 277 aa with a calculated M(r) of 30,304. Expression studies showed that the cpo from P. pyrrocinia is functionally expressed in Escherichia coli and Streptomyces lividans. Based on the overproduction of the CPO in E. coli, a novel and simple purification procedure was developed allowing the isolation of about 800-fold more CPO per gram of cells than was originally isolated from P. pyrrocinia. Comparison with the aa sequence of the bromoperoxidase BPO-A2 from S. aureofaciens ATCC10762 revealed an identity of 38%.

摘要

测定了包含氯过氧化物酶（CPO）编码基因（cpo）及其侧翼区域的1.5 kb吡咯菌素假单胞菌DNA片段的核苷酸序列。cpo编码一个由278个氨基酸（aa）组成的蛋白质。成熟酶不含N端甲硫氨酸，因此CPO单体由277个aa组成，计算的M（r）为30304。表达研究表明，来自吡咯菌素假单胞菌的cpo在大肠杆菌和淡紫链霉菌中功能性表达。基于CPO在大肠杆菌中的过量表达，开发了一种新颖且简单的纯化方法，每克细胞可分离出比最初从吡咯菌素假单胞菌中分离出的CPO多约800倍的CPO。与来自金黄色链霉菌ATCC10762的溴过氧化物酶BPO-A2的aa序列比较显示同一性为38%。