title: Functional robustness of a polycyclic aromatic hydrocarbon metabolic network examined in a nidA aromatic ring-hydroxylating oxygenase mutant of Mycobacterium vanbaalenii PYR-1.
abstract: In this study, we obtained over 4,000 transposon mutants of Mycobacterium vanbaalenii PYR-1 and analyzed one of the mutants, 8F7, which appeared to lose its ability to degrade pyrene while still being able to degrade fluoranthene. This mutant was identified to be defective in nidA, encoding an aromatic ring-hydroxylating oxygenase (RHO), known to be involved in the initial oxidation step of pyrene degradation. When cultured with pyrene as a sole source of polycyclic aromatic hydrocarbon (PAH), high-pressure liquid chromatography analysis revealed that the nidA mutant showed a significant decrease in the rate of pyrene degradation compared to the wild-type PYR-1, although pyrene was still being degraded. However, when incubated with PAH mixtures including pyrene, phenanthrene, and fluoranthene, the pyrene degradation rate of the mutant was higher than that of the mutant previously incubated with pyrene as a sole source of PAH. There was no significant difference between wild-type PYR-1 and the mutant in the rates of phenanthrene and fluoranthene degradation. From the whole-cell proteome analysis of mutant 8F7 induced by pyrene, we identified expression of a number of RHO enzymes which are suspected to be responsible for pyrene degradation in the nidA mutant, which had no expression of NidA. Taken together, results in this study provide direct evidence for the in vivo functional role of nidA in pyrene degradation at the level of the ring-cleavage-process (RCP) functional module but also for the robustness of the PAH metabolic network (MN) to such a genetic perturbation.
authors.affiliation: Division of Microbiology, National Center for Toxicological Research/U.S. FDA, Jefferson, Arkansas, USA.
authors.forename: Seong-Jae
authors.initials: SJ
authors.lastname: Kim
authors.forename: Jaekyeong
authors.initials: J
authors.lastname: Song
authors.forename: Ohgew
authors.initials: O
authors.lastname: Kweon
authors.forename: Ricky D
authors.initials: RD
authors.lastname: Holland
authors.forename: Dae-Wi
authors.initials: DW
authors.lastname: Kim
authors.forename: Jongnam
authors.initials: J
authors.lastname: Kim
authors.forename: Li-Rong
authors.initials: LR
authors.lastname: Yu
authors.forename: Carl E
authors.initials: CE
authors.lastname: Cerniglia
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis; D003470:Culture Media / Q000737:chemistry; D004251:DNA Transposable Elements; D017353:Gene Deletion; D020869:Gene Expression Profiling; D053858:Metabolic Networks and Pathways; D016254:Mutagenesis, Insertional; D009161:Mycobacterium / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D010105:Oxygenases / Q000235:genetics*; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism*; D020543:Proteome / Q000032:analysis
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D004251:DNA Transposable Elements; D011084:Polycyclic Aromatic Hydrocarbons; D020543:Proteome; D010105:Oxygenases