Open AccessCloning of the wide spectrum amidase gene from Brevibacterium sp. R312 by genetic complementation. Overexpression in Brevibacterium sp. and Escherichia coli
Author(s) -
Azza S.,
Bigey F.,
Arnaud A.,
Galzy P.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb07155.x
Subject(s) - brevibacterium , subcloning , complementation , biology , gene , escherichia coli , plasmid , mutant , microbiology and biotechnology , amidase , cosmid , cloning (programming) , genetics , bacteria , microorganism , computer science , programming language
The amiE gene of Brevibacterium sp. R312 encoding wide spectrum amidase was isolated by complementation of a Brevibacterium sp. mutant using a plasmid gene bank of chromosomal DNA. The amiE structural gene and its promoter were localized on a 1.8‐kb fragment by subsequent subcloning and complementation studies. In Brevibacterium sp., the investigation of amidase activities related to one copy of the gene suggested that the regulation of the amiE gene expression was under negative control. High expression levels have been obtained in Brevibacterium sp. and, after substitution of the amiE promoter by the tac promoter, in Escherichia coli .