title: Expression in Escherichia coli of biphenyl 2,3-dioxygenase genes from a Gram-positive polychlorinated biphenyl degrader, Rhodococcus jostii RHA1.
abstract: Rhodococcus jostii RHA1 is a polychlorinated biphenyl degrader. Multi-component biphenyl 2,3-dioxygenase (BphA) genes of RHA1 encode large and small subunits of oxygenase component and ferredoxin and reductase components. They did not express enzyme activity in Escherichia coli. To obtain BphA activity in E. coli, hybrid BphA gene derivatives were constructed by replacing ferredoxin and/or reductase component genes of RHA1 with those of Pseudomonas pseudoalcaligenes KF707. The results obtained indicate a lack of catalytic activity of the RHA1 ferredoxin component gene, bphAc in E. coli. To determine the cause of inability of RHA1 bphAc to express in E. coli, the bphAc gene was introduced into Rosetta (DE3) pLacI, which has extra tRNA genes for rare codons in E. coli. The resulting strain abundantly produced the bphAc product, and showed activity. These results suggest that codon usage bias is involved in inability of RHA1 bphAc to express its catalytic activity in E. coli.
authors.affiliation: Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka-cho, Nagaoka, Niigata 940-2188, Japan.
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Ohmori
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Morita
authors.forename: Megumi
authors.initials: M
authors.lastname: Tanaka
authors.forename: Masanori
authors.initials: M
authors.lastname: Tomoi
authors.forename: Keisuke
authors.initials: K
authors.lastname: Miyauchi
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kasai
authors.forename: Kensuke
authors.initials: K
authors.lastname: Furukawa
authors.forename: Eiji
authors.initials: E
authors.lastname: Masai
authors.forename: Masao
authors.initials: M
authors.lastname: Fukuda
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011078:Polychlorinated Biphenyls / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D012343:RNA, Transfer / Q000235:genetics; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics*
chemical_list: D003062:Codon; D007506:Iron-Sulfur Proteins; D012343:RNA, Transfer; D011078:Polychlorinated Biphenyls; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase