Cloning and expression of the Arthrobacter globiformis KZT1 fcbA gene encoding dehalogenase (4-chlorobenzoate-4-hydroxylase) in Escherichia coli.
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 Mr = 58 kDa (and/or Mr = 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/l to 0.7 g/l as well as a sudden reduction in the reaction efficiency at higher substrate concentrations were observed.
Tsoi
T V
TV
Institute of Biochemistry and Physiology of Microorganisms, U.S.S.R. Academy of Sciences, Moscow.
Zaitsev
G M
GM
Plotnikova
E G
EG
Kosheleva
I A
IA
Boronin
A M
AM
D001173:Arthrobacter / Q000201:enzymology / Q000235:genetics*; D002723:Chlorobenzoates / Q000378:metabolism*; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic
D002723:Chlorobenzoates; D003470:Culture Media; D006867:Hydrolases; C042846:4-chlorobenzoate dehalogenase; C040768:4-chlorobenzoic acid