title: Identification of novel extracellular protein for PCB/biphenyl metabolism in Rhodococcus jostii RHA1.
abstract: Rhodococcus jostii RHA1 (RHA1) degrades polychlorinated biphenyl (PCB) via co-metabolism with biphenyl. To identify the novel open reading frames (ORFs) that contribute to PCB/biphenyl metabolism in RHA1, we compared chromatin immunoprecipitation chip and transcriptomic data. Six novel ORFs involved in PCB/biphenyl metabolism were identified. Gene deletion mutants of these 6 ORFs were made and were tested for their ability to grow on biphenyl. Interestingly, only the ro10225 deletion mutant showed deficient growth on biphenyl. Analysis of Ro10225 protein function showed that growth of the ro10225 deletion mutant on biphenyl was recovered when exogenous recombinant Ro10225 protein was added to the culture medium. Although Ro10225 protein has no putative secretion signal sequence, partially degraded Ro10225 protein was detected in conditioned medium from wild-type RHA1 grown on biphenyl. This Ro10225 fragment appeared to form a complex with another PCB/biphenyl oxidation enzyme. These results indicated that Ro10225 protein is essential for the formation of the PCB/biphenyl dioxygenase complex in RHA1.
authors.affiliation: a Division of Chemistry and Biochemistry, Department of Biotechnology , Graduate School of Natural Science and Technology, Okayama University , Okayama , Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Atago
authors.affiliation: b Department of Bioengineering , Nagaoka University of Technology , Nagaoka , Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Shimodaira
authors.affiliation: b Department of Bioengineering , Nagaoka University of Technology , Nagaoka , Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Araki
authors.affiliation: d Department of Mechanical Precision Engineering , Malaysia-Japan International Institute of Technology, Universiti Teknologi Malaysia , Kuala Lumpur , Malaysia.
authors.forename: Nor'azizi
authors.initials: N
authors.lastname: Bin Othman
authors.affiliation: c Department of Environmental Engineering and Green Technology , Malaysia-Japan International Institute of Technology, Universiti Teknologi Malaysia , Kuala Lumpur , Malaysia.
authors.forename: Zuriati
authors.initials: Z
authors.lastname: Zakaria
authors.affiliation: b Department of Bioengineering , Nagaoka University of Technology , Nagaoka , Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Fukuda
authors.affiliation: a Division of Chemistry and Biochemistry, Department of Biotechnology , Graduate School of Natural Science and Technology, Okayama University , Okayama , Japan.
authors.forename: Junichiro
authors.initials: J
authors.lastname: Futami
authors.affiliation: c Department of Environmental Engineering and Green Technology , Malaysia-Japan International Institute of Technology, Universiti Teknologi Malaysia , Kuala Lumpur , Malaysia.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hara
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001713:Biphenyl Compounds / Q000378:metabolism* / Q000494:pharmacology; D047369:Chromatin Immunoprecipitation; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned / Q000737:chemistry; D049308:Dioxygenases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D058977:Molecular Sequence Annotation; D016366:Open Reading Frames; D011078:Polychlorinated Biphenyls / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012240:Rhodococcus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D059467:Transcriptome
chemical_list: D001426:Bacterial Proteins; D001713:Biphenyl Compounds; D017077:Culture Media, Conditioned; D049308:Dioxygenases; D011078:Polychlorinated Biphenyls; D011994:Recombinant Proteins; C010574:biphenyl
keywords: RHA1; Rhodococcus jostii; biodegradation; biphenyl; polychlorinated biphenyl (PCB)