Proposed involvement of a soluble methane monooxygenase homologue in the cyclohexane-dependent growth of a new Brachymonas species.
High-throughput mRNA differential display (DD) was used to identify genes induced by cyclohexane in Brachymonas petroleovorans CHX, a recently isolated beta-proteobacterium that grows on cyclohexane. Two metabolic gene clusters were identified multiple times in independent reverse transcription polymerase chain reactions (RT-PCR) in the course of this DD experiment. These clusters encode genes believed to be required for cyclohexane metabolism. One gene cluster (8 kb) encodes the subunits of a multicomponent hydroxylase related to the soluble butane of Pseudomonas butanovora and methane monooxygenases (sMMO) of methanotrophs. We propose that this butane monooxygenase homologue carries out the oxidation of cyclohexane into cyclohexanol during growth. A second gene cluster (11 kb) contains almost all the genes required for the oxidation of cyclohexanol to adipic acid. Real-time PCR experiments confirmed that genes from both clusters are induced by cyclohexane. The role of the Baeyer-Villiger cyclohexanone monooxygenase of the second cluster was confirmed by heterologous expression in Escherichia coli.
Brzostowicz
Patricia C
PC
E. I. Dupont de Nemours and Company, Central Research and Development, Wilmington, DE 19880-0328, USA.
Walters
Dana M
DM
Jackson
Raymond E
RE
Halsey
Kimberly H
KH
Ni
Hao
H
Rouvière
Pierre E
PE
D000272:Adipates / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D042621:Comamonadaceae / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D003510:Cyclohexanes / Q000378:metabolism*; D003511:Cyclohexanols / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006899:Mixed Function Oxygenases / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000235:genetics* / Q000502:physiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000235:genetics; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
D000272:Adipates; D001426:Bacterial Proteins; D003510:Cyclohexanes; D003511:Cyclohexanols; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012333:RNA, Messenger; D011994:Recombinant Proteins; C029900:adipic acid; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C021555:methane monooxygenase