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PMID:11673430

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Citation

Kitagawa, W, Miyauchi, K, Masai, E and Fukuda, M (2001) Cloning and characterization of benzoate catabolic genes in the gram-positive polychlorinated biphenyl degrader Rhodococcus sp. strain RHA1. J. Bacteriol. 183:6598-606

Abstract

Benzoate catabolism is thought to play a key role in aerobic bacterial degradation of biphenyl and polychlorinated biphenyls (PCBs). Benzoate catabolic genes were cloned from a PCB degrader, Rhodococcus sp. strain RHA1, by using PCR amplification and temporal temperature gradient electrophoresis separation. A nucleotide sequence determination revealed that the deduced amino acid sequences encoded by the RHA1 benzoate catabolic genes, benABCDK, exhibit 33 to 65% identity with those of Acinetobacter sp. strain ADP1. The gene organization of the RHA1 benABCDK genes differs from that of ADP1. The RHA1 benABCDK region was localized on the chromosome, in contrast to the biphenyl catabolic genes, which are located on linear plasmids. Escherichia coli cells containing RHA1 benABCD transformed benzoate to catechol via 2-hydro-1,2-dihydroxybenzoate. They transformed neither 2- nor 4-chlorobenzoates but did transform 3-chlorobenzoate. The RHA1 benA gene was inactivated by insertion of a thiostrepton resistance gene. The resultant mutant strain, RBD169, neither grew on benzoate nor transformed benzoate, and it did not transform 3-chlorobenzoate. It did, however, exhibit diminished growth on biphenyl and growth repression in the presence of a high concentration of biphenyl (13 mM). These results indicate that the cloned benABCD genes could play an essential role not only in benzoate catabolism but also in biphenyl catabolism in RHA1. Six rhodococcal benzoate degraders were found to have homologs of RHA1 benABC. In contrast, two rhodococcal strains that cannot transform benzoate were found not to have RHA1 benABC homologs, suggesting that many Rhodococcus strains contain benzoate catabolic genes similar to RHA1 benABC.

Links

PubMed PMC95491 Online version:10.1128/JB.183.22.6598-6606.2001

Keywords

Base Sequence; Benzoates/metabolism; Chromosomes, Bacterial; Cloning, Molecular; Dioxygenases; Genes, Bacterial; Iron-Sulfur Proteins; Models, Chemical; Molecular Sequence Data; Mutagenesis, Insertional; Mutation; Oxidoreductases/genetics; Oxidoreductases/metabolism; Oxygenases/genetics; Oxygenases/metabolism; Polychlorinated Biphenyls/metabolism; Rhodococcus/enzymology; Rhodococcus/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RHOJR:Q932V1

GO:0043639: benzoate catabolic process

ECO:0000315:

P

Fig. 6 indicates that the wildtype RHA1 is able to degrade benzoate, a product of biphenyl metabolism, as indicated by a final concentration of 0mM benzoate in the medium. The mutant, RDB169 (benA deleted), is unable to degrade the benzoate as effectively as shown by final benzoate concentrations between 3.3 and 6.8mM in the medium.

complete
CACAO 4287

RHOJR:Q932V1

involved_in

GO:0043639: benzoate catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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