Gene mdpC plays a regulatory role in the methyl-tert-butyl ether degradation pathway of Methylibium petroleiphilum strain PM1

Abstract

Among the few well-studied bacteria known to utilize methyl tert-butyl ether (MTBE) as a sole carbon source, Methylibium petroleiphilum PM1 is a well-characterized organism with a sequenced genome; however, knowledge of the genetic regulation of its MTBE degradation pathway is limited. We investigated the role of a putative transcriptional activator gene, mdpC, in the induction of MTBE degradation genes mdpA (encoding MTBE monooxygenase) and mdpJ (encoding tert-butyl alcohol hydroxylase) of strain PM1 in a gene-knockout mutant mdpC−. We also utilized quantitative reverse transcriptase PCR (qRT-PCR) assays targeting genes mdpA, mdpJ and mdpC to determine the effects of the mutation on transcription of these genes. Our results indicate that gene mdpC is involved in the induction of both mdpA and mdpJ in response to MTBE and tert-butyl alcohol (TBA) exposure in PM1. An additional independent mechanism may be involved in the induction of mdpJ in the presence of TBA.

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“…The q-PCR program was 5 s at 95 °C, 34 s at 60 °C, and then 40 cycles of 30 s at 95 °C with melt curve added. The mRNA level of DNA gyrase subunit B (GyrB) was used as an internal control . The primers of target genes are listed in Table S2.…”

“…The mRNA level of DNA gyrase subunit B (GyrB) was used as an internal control. 23 The primers of target genes are listed in Table S2.…”