title: Identification of a gene involved in plasmid structural instability in Corynebacterium glutamicum.
abstract: Expression plasmids that facilitate production of bio-based products are susceptible to toxic effects that frequently affect plasmid structural stability in recombinant microbial cells. In order to enhance plasmid stability in recombinant Corynebacterium glutamicum, an expression plasmid containing genes of the Clostridium acetobutylicum butyryl-CoA synthesis operon with high structural instability within wild-type C. glutamicum was employed. From a total of 133 mutants exhibiting disruptions in 265 suspect genes, only cgR_0322-deficient mutant was able to maintain the expression plasmid intact. The mutant exhibited normal growth under standard laboratory conditions but its transformation efficiency was about one order of magnitude lower than that of wild-type strain. The cgR_0322 gene encodes an endonuclease that is active against single- as well as double-stranded DNA substrates in the presence of Mg(2+). The cgR_0322-deficient strain should therefore facilitate the development of more robust C. glutamicum strains to be used as microbial production hosts.
authors.affiliation: Molecular Microbiology and Biotechnology Group, Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto, 619-0292, Japan.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Kitade
authors.forename: Shohei
authors.initials: S
authors.lastname: Okino
authors.forename: Wataru
authors.initials: W
authors.lastname: Gunji
authors.forename: Kazumi
authors.initials: K
authors.lastname: Hiraga
authors.forename: Masako
authors.initials: M
authors.lastname: Suda
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Suzuki
authors.forename: Masayuki
authors.initials: M
authors.lastname: Inui
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yukawa
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004720:Endonucleases