Molecular cloning of a novel gene, dtsR, which rescues the detergent sensitivity of a mutant derived from Brevibacterium lactofermentum.
Several strains of Corynebacterium and Brevibacterium are known for their ability to secrete large amounts of amino acids, especially L-glutamate. We focused on the mechanism of L-glutamate secretion triggered by a detergent, namely polyoxyethylenesorbitan monopalmitate (PESP). A mutant strain, AJ11060, derived from Brevibacterium lactofermentum ATCC 13869 indicates the sensitivity to PESP. A multicopy suppresser gene that compliments the sensitivity of AJ11060 to the detergent was derived from a gene library of B. lactofermentum AJ12036. A 2855-bp DNA fragment was cloned and sequenced. An open reading frame was found that coded for the rescuer gene of the sensitivity to PESP of AJ11060 and was designated dtsR. The expression of the dtsR gene in B. lactofermentum was confirmed by using anti-DtsR antibody. The deduced DtsR protein indicated significant homology with some biotin enzymes such as the beta chain of propionyl-CoA carboxylase from rat (48.3%) and human (48.7%), or a 12S chain of methylmalonyl-CoA carboxyltransferase from Propionibacterium freudenreichii (43.1%).
Kimura
E
E
Technology and Engineering Laboratories, Ajinomoto Co., Inc., Kanagawa, Japan.
Abe
C
C
Kawahara
Y
Y
Nakamatsu
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D001483:Base Sequence; D001710:Biotin / Q000737:chemistry; D001951:Brevibacterium / Q000235:genetics*; D003001:Cloning, Molecular; D003902:Detergents / Q000494:pharmacology*; D005798:Genes, Bacterial; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation; D011136:Polysorbates / Q000494:pharmacology*; D051381:Rats
D001426:Bacterial Proteins; D003902:Detergents; D011136:Polysorbates; D018698:Glutamic Acid; D001710:Biotin