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J. Bacteriol. doi:10.1128/JB.00128-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

A Polyomic Approach to Elucidate the Fluoranthene Degradative Pathway in Mycobacterium vanbaalenii PYR-1

Division of Microbiology, Division of Systems Toxicology, Division of Biochemical Toxicology, National Center for Toxicological Research/U.S. FDA, Jefferson, AR 72079, USA

^* To whom correspondence should be addressed. Email: carl.cerniglia{at}hhs.fda.gov.

	Abstract

Mycobacterium vanbaalenii PYR-1 is capable of degrading a wide range of high-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs), including fluoranthene. We conducted a combination of metabolism, genomic, and proteomic technologies to investigate fluoranthene degradation in this strain. Thirty-seven fluoranthene metabolites including potential isomers were isolated from the culture medium and analyzed by high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and UV-visible absorption. Total proteins were separated by one-dimensional (1D) gel and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in conjunction with the M. vanbaalenii PYR-1 genome sequence (http://jgi.doe.gov), which resulted in the identification of 1122 proteins. Among them, 53 enzymes were determined as likely involved in fluoranthene degradation. We integrated the metabolic information with the genomic and proteomic results and proposed pathways for the degradation of fluoranthene. According to our hypothesis, the oxidation of fluoranthene is initiated by dioxygenation at the C-1,2, C-2,3, and C-7,8 positions. The C-1,2 and C-2,3 dioxygenation routes degrade fluoranthene via fluorene-type metabolites, whereas the C-7,8 routes oxidize fluoranthene via acenaphthylene-type metabolites. The major site of dioxygenation is the C-2,3 dioxygenation route, which consists of 18 enzymatic steps via 9-fluorenone-1-carboxylic acid and phthalate with the initial ring-hydroxylating oxygenase, NidA3B3, oxidizing fluoranthene to fluoranthene cis-2,3-dihydrodiol. Non-specific monooxygenation of fluoranthene with subsequent O-methylation of dihydroxyfluoranthene also occurs as a detoxification reaction.