Involvement of BmoR and BmoG in n-alkane metabolism in 'Pseudomonas butanovora'.
'Pseudomonas butanovora' uses an alcohol-inducible alkane monooxygenase (BMO) to grow on C(2)-C(9) n-alkanes. Five ORFs were identified flanking the BMO structural genes. Two of the ORFs, bmoR, encoding a putative sigma(54)-transcriptional regulator BmoR, and bmoG, encoding a putative GroEL chaperonin BmoG, were analysed by gene-inactivation experiments. The BmoR-deficient mutant grew at slower growth rates than the wild-type on C(2)-C(5) n-alkanes and showed little to no growth on C(6)-C(8) n-alkanes within 7 days. A BmoR-deficient mutant was constructed in the 'P. butanovora' bmoX : : lacZ reporter strain and used to test whether bmoR was involved in bmoX induction after growth on C(2)-C(8) carbon sources. In acetate- or lactate-grown cells, C(2)-C(8) n-alcohols failed to induce beta-galactosidase activity. In contrast, in propionate-, butyrate- or pentanoate-grown cells, n-butanol induced approximately 45 % of the beta-galactosidase activity observed in the control bmoX : : lacZ strain. In propionate-grown cells, C(2)-C(5) n-alcohols induced beta-galactosidase activity, whereas C(7) and C(8) n-alcohols did not. BmoR may act as a sigma(54)-transcriptional regulator of bmo that is controlled by the n-alcohol produced in the alkane oxidation. During growth on short-chain-length fatty acids, however, another BMO regulatory system seems to be activated to promote transcription of bmo by short-chain-length alcohols (i.e. </=C(6)). The bmoG-deficient mutant did not grow on C(2)-C(8) n-alkanes; however, it was capable of transcribing bmoX and bmoC of the BMO operon. BmoG may act as a chaperonin to assemble competent BMO.
Kurth
Elizabeth G
EG
Department of Botany and Plant Pathology, 2082 Cordley Hall, Oregon State University, Corvallis, OR 97331, USA.
Doughty
David M
DM
Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA.
Bottomley
Peter J
PJ
Department of Crop and Soil Science, Oregon State University, Corvallis, OR 97331, USA.|Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA.
Arp
Daniel J
DJ
Department of Botany and Plant Pathology, 2082 Cordley Hall, Oregon State University, Corvallis, OR 97331, USA.
Sayavedra-Soto
Luis A
LA
Department of Botany and Plant Pathology, 2082 Cordley Hall, Oregon State University, Corvallis, OR 97331, USA.
D000085:Acetates / Q000378:metabolism; D000438:Alcohols / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism; D018833:Chaperonins / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005232:Fatty Acids, Volatile / Q000378:metabolism; D017353:Gene Deletion; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D007773:Lactates / Q000378:metabolism; D008954:Models, Biological; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011422:Propionates / Q000378:metabolism; D011549:Pseudomonas / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D000085:Acetates; D000438:Alcohols; D000473:Alkanes; D002087:Butyrates; D004269:DNA, Bacterial; D005232:Fatty Acids, Volatile; D007773:Lactates; D018832:Molecular Chaperones; D011422:Propionates; D012808:Sigma Factor; D014157:Transcription Factors; D001616:beta-Galactosidase; D018833:Chaperonins