Identification of a Novel Dioxygenase Involved in Metabolism of<i>o</i>-Xylene, Toluene, and Ethylbenzene by<i>Rhodococcus</i>sp. Strain DK17

Kim, Dockyu; Chae, Jong‐Chan; Zylstra, Gerben J.; Kim, Youngsoo; Kim, Seong‐Ki; Nam, Myung Hee; Kim, Young Min; Kim, Eungbin

doi:10.1128/aem.70.12.7086-7092.2004

Cited by 54 publications

(39 citation statements)

References 28 publications

(36 reference statements)

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“…Strain DK17 possesses three linear megaplasmids (380-kb pDK1, 330-kb pDK2, and 750-kb pDK3). The genes for alkylbenzene degradation are present on pDK2, while the gene clusters for phthalate degradation are duplicated and found on both pDK2 and pDK3 (2,4).…”

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Draft Genome Sequence and Comparative Analysis of the Superb Aromatic-Hydrocarbon Degrader Rhodococcus sp. Strain DK17

et al. 2012

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Draft Genome Sequence and Comparative Analysis of the Superb Aromatic-Hydrocarbon Degrader Rhodococcus sp. Strain DK17

et al. 2012

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“…strain C125, and Rhodococcus sp. strain DK17 belong to this class (13,16,30). These strains are thought to metabolize o-xylene via 3,4-dimethylcatechol.…”

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“…o-Xylene dioxygenase genes (akbA1 a , akbA2 a , and akbA3) have recently been isolated from Rhodococcus sp. strain DK17 (16), which can grow on o-xylene but cannot grow on m-or p-xylene (17). The second pathway for o-xylene degradation is one initiated by a ring-hydroxylating monooxygenase (Fig.…”

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Isolation and Characterization of o -Xylene Oxygenase Genes from Rhodococcus opacus TKN14

Maruyama

Ishikura

Taki

et al. 2005

Appl Environ Microbiol

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o-Xylene is one of the most difficult-to-degrade environmental pollutants. We report here Rhodococcus genes mediating oxygenation in the first step of o-xylene degradation. Rhodococcus opacus TKN14, isolated from soil contaminated with o-xylene, was able to utilize o-xylene as the sole carbon source and to metabolize it to o-methylbenzoic acid. A cosmid library from the genome of this strain was constructed in Escherichia coli. A bioconversion analysis revealed that a cosmid clone incorporating a 15-kb NotI fragment had the ability to convert o-xylene into o-methylbenzyl alcohol. The sequence analysis of this 15-kb region indicated the presence of a gene cluster significantly homologous to the naphthalene-inducible dioxygenase gene clusters (nidABCD) that had been isolated from Rhodococcus sp. strain I24. Complementation studies, using E. coli expressing various combinations of individual open reading frames, revealed that a gene (named nidE) for rubredoxin (Rd) and a novel gene (named nidF) encoding an auxiliary protein, which had no overall homology with any other proteins, were indispensable for the methyl oxidation reaction of o-xylene, in addition to the dioxygenase iron-sulfur protein genes (nidAB). Regardless of the presence of NidF, the enzyme composed of NidABE was found to function as a typical naphthalene dioxygenase for converting naphthalene and various (di)methylnaphthalenes into their corresponding cis-dihydrodiols. All the nidABEF genes were transcriptionally induced in R. opacus TKN14 by the addition of o-xylene to a mineral salt medium. It is very likely that these genes are involved in the degradation pathways of a wide range of aromatic hydrocarbons by Rhodococcus species as the first key enzyme. o-, m-and p-xylenes, which are included in petroleum, are used as solvents in the production of chemicals and drugs, as thinners for paints and enamels, and as intermediates for the synthesis of numerous chemical compounds. They are therefore widespread environmental contaminants in groundwater and soil (4,11,27). In m-and p-xylenes, numerous bacteria that are able to utilize them as the sole carbon and energy source have been isolated, and details of their catabolic pathways have been well documented. The most elucidated examples are the catabolism of toluene, m-xylene, and p-xylene with the enzymes encoded on a transmissible TOL plasmid (pWW0) from Pseudomonas putida mt-2 (38, 39). Xylene monooxygenase is the first enzyme on this degradation pathway (14, 34). This enzyme consists of two polypeptide subunits encoded by the xylM and xylA genes. XylA, the NADH acceptor reductase component (34), has been characterized as an electron transport protein transferring reducing equivalents from NADH to XylM, which is the hydroxylase component located in the membrane (18). Toluene and m-and p-xylenes are, respectively, metabolized with XylMA to benzyl alcohol and m-and p-methylbenzyl alcohol. These respective alcohols are further metabolized to benzoate and m-and p-toluates via their corresponding aldehydes b...

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“…strain DK17 was originally isolated for the ability to grow on o-xylene and has the capability to utilize such aromatic compounds as benzene, alkylbenzenes (toluene, ethylbenzene, isopropylbenzene, and n-propyl-to n-hexylbenzenes), phenol, and phthalates as sole carbon and energy sources (2,14,15,16,17). This catabolic versatility led us to investigate the adaptability of DK17 to the presence of multiple aromatic hydrocarbons as carbon and energy sources.…”

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Benzoate Catabolite Repression of the Phthalate Degradation Pathway in Rhodococcus sp. Strain DK17

Choi

Zylstra

Kim

2007

Appl Environ Microbiol

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Rhodococcus sp. strain DK17 exhibits a catabolite repression-like response when provided simultaneously with benzoate and phthalate as carbon and energy sources. Benzoate in the medium is depleted to detection limits before the utilization of phthalate begins. The transcription of the genes encoding benzoate and phthalate dioxygenase paralleled the substrate utilization profile. Two mutant strains with defective benzoate dioxygenases were unable to utilize phthalate in the presence of benzoate, although they grew normally on phthalate in the absence of benzoate.

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Identification of a Novel Dioxygenase Involved in Metabolism ofo-Xylene, Toluene, and Ethylbenzene byRhodococcussp. Strain DK17

Cited by 54 publications

References 28 publications

Draft Genome Sequence and Comparative Analysis of the Superb Aromatic-Hydrocarbon Degrader Rhodococcus sp. Strain DK17

Draft Genome Sequence and Comparative Analysis of the Superb Aromatic-Hydrocarbon Degrader Rhodococcus sp. Strain DK17

Isolation and Characterization of o -Xylene Oxygenase Genes from Rhodococcus opacus TKN14

Benzoate Catabolite Repression of the Phthalate Degradation Pathway in Rhodococcus sp. Strain DK17

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