Open AccessCloning and expression of the Arthrobacter globiformis KZT1 fcbA gene encoding dehalogenase (4‐chlorobenzoate‐4‐hydroxylase) in Escherichia coli
Author(s) -
Tsoi Tamara V.,
Zaitsev Gennady M.,
Plotnikova Elena G.,
Kosheleva Irina A.,
Boronin Alexander M.
Publication year - 1991
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.1574-6968.1991.tb04741.x
Subject(s) - escherichia coli , cloning (programming) , arthrobacter , gene , bacteria , enterobacteriaceae , molecular cloning , dehalogenase , recombinant dna , biology , gene product , microbiology and biotechnology , gene expression , biochemistry , enzyme , lac operon , chemistry , genetics , computer science , programming language
The fsbA gene controlling the first step of 4‐chlorobenzoic acid (4CBA) metabolism in the Gram‐positive soil bacterium Arthrobacter globiformis KZT1 has been cloned and analysed in Escherichia coli . The E. coli minicells analysis showed that a polypeptide(s) with M r = 58 kDa (and/or M r = 32 kDa) can be the fcbA product(s). Despite the gene dose amplification and control of the E. coli inducible Plac promoter, the level of functional expression of the fcbA gene in E. coli cells seems comparable only with that in the parental KZT1 strain. Effective 4CBA dechlorination by recombinant cells during growth in the presence of substrate within a range of concentrations 0.1 g/1 to 0.7 g/1 as well as a sudden reduction in the reaction efficiency at higher substrate concentrations were observed.