title: SmoXYB1C1Z of Mycobacterium sp. strain NBB4: a soluble methane monooxygenase (sMMO)-like enzyme, active on C2 to C4 alkanes and alkenes.
abstract: Monooxygenase (MO) enzymes initiate the aerobic oxidation of alkanes and alkenes in bacteria. A cluster of MO genes (smoXYB1C1Z) of thus-far-unknown function was found previously in the genomes of two Mycobacterium strains (NBB3 and NBB4) which grow on hydrocarbons. The predicted Smo enzymes have only moderate amino acid identity (30 to 60%) to their closest homologs, the soluble methane and butane MOs (sMMO and sBMO), and the smo gene cluster has a different organization from those of sMMO and sBMO. The smoXYB1C1Z genes of NBB4 were cloned into pMycoFos to make pSmo, which was transformed into Mycobacterium smegmatis mc(2)-155. Cells of mc(2)-155(pSmo) metabolized C2 to C4 alkanes, alkenes, and chlorinated hydrocarbons. The activities of mc(2)-155(pSmo) cells were 0.94, 0.57, 0.12, and 0.04 nmol/min/mg of protein with ethene, ethane, propane, and butane as substrates, respectively. The mc(2)-155(pSmo) cells made epoxides from ethene, propene, and 1-butene, confirming that Smo was an oxygenase. Epoxides were not produced from larger alkenes (1-octene and styrene). Vinyl chloride and 1,2-dichloroethane were biodegraded by cells expressing Smo, with production of inorganic chloride. This study shows that Smo is a functional oxygenase which is active against small hydrocarbons. M. smegmatis mc(2)-155(pSmo) provides a new model for studying sMMO-like monooxygenases.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, Australia.
authors.forename: Kiri E
authors.initials: KE
authors.lastname: Martin
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, Australia.
authors.forename: Jazmin
authors.initials: J
authors.lastname: Ozsvar
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, Australia nicholas.coleman@sydney.edu.au.
authors.forename: Nicholas V
authors.initials: NV
authors.lastname: Coleman
mesh_terms: D000473:Alkanes / Q000378:metabolism*; D000475:Alkenes / Q000378:metabolism*; D001711:Biotransformation; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004852:Epoxy Compounds / Q000378:metabolism; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D009161:Mycobacterium / Q000201:enzymology* / Q000235:genetics; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D014169:Transformation, Bacterial
chemical_list: D000473:Alkanes; D000475:Alkenes; D004269:DNA, Bacterial; D004852:Epoxy Compounds; D006843:Hydrocarbons, Chlorinated; D010105:Oxygenases; C021555:methane monooxygenase