Identification of novel extracellular protein for PCB/biphenyl metabolism in Rhodococcus jostii RHA1

Bioscience, Biotechnology, and Biochemistry
Yuki AtagoHirofumi Hara

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.

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Citations

Jan 29, 2019·The Journal of General and Applied Microbiology·Shazwana Sha'araniZuriati Zakaria

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Datasets Mentioned

BETA
GPL3918

Methods Mentioned

BETA
PCR
immunoprecipitation
gel filtration
dot blotting
ChIP-chip
dot-blot

Software Mentioned

ImaGene
BLAST
Array Pipe

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