title: Untangling the multiple monooxygenases of Mycobacterium chubuense strain NBB4, a versatile hydrocarbon degrader.
abstract: Mycobacterium strain NBB4 was isolated on ethene as part of a bioprospecting study searching for novel monooxygenase (MO) enzymes of interest to biocatalysis and bioremediation. Previous work indicated that strain NBB4 contained an unprecedented diversity of MO genes, and we hypothesized that each MO type would support growth on a distinct hydrocarbon substrate. Here, we attempted to untangle the relationships between MO types and hydrocarbon substrates. Strain NBB4 was shown to grow on C2 -C4 alkenes and C2 -C16 alkanes. Complete gene clusters encoding six different monooxygenases were recovered from a fosmid library, including homologues of ethene MO (etnABCD), propene MO (pmoABCD), propane MO (smoABCD), butane MO (smoXYB1C1Z), cytochrome P450 (CYP153; fdx-cyp-fdr) and alkB (alkB-rubA1-rubA2). Catabolic enzymes involved in ethene assimilation (EtnA, EtnC, EtnD, EtnE) and alkane assimilation (alcohol and aldehyde dehydrogenases) were identified by proteomics, and we showed for the first time that stress response proteins (catalase/peroxidase, chaperonins) were induced by growth on C2 -C5 alkanes and ethene. Surprisingly, none of the identified MO genes could be specifically associated with oxidation of small alkanes, and thus the nature of the gaseous alkane MO in NBB4 remains mysterious.
authors.affiliation: School of Molecular Bioscience, Building G08, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: Nicholas V
authors.initials: NV
authors.lastname: Coleman
authors.forename: Sheree
authors.initials: S
authors.lastname: Yau
authors.forename: Neil L
authors.initials: NL
authors.lastname: Wilson
authors.forename: Laura M
authors.initials: LM
authors.lastname: Nolan
authors.forename: Margaret D
authors.initials: MD
authors.lastname: Migocki
authors.forename: Mai-Anh
authors.initials: MA
authors.lastname: Ly
authors.forename: Ben
authors.initials: B
authors.lastname: Crossett
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Holmes