1887

Abstract

The key enzyme in methane metabolism is methane monooxygenase (MMO), which catalyses the oxidation of methane to methanol. Some methanotrophs, including (Bath), possess two distinct MMOs. The level of copper in the environment regulates the biosynthesis of the MMO enzymes in these methanotrophs. Under low-copper conditions, soluble MMO (sMMO) is expressed and regulation takes place at the level of transcription. The structural genes of sMMO were previously identified as , and . Putative transcriptional start sites, containing a - and a -dependent motif, were identified in the 5′ region of . The promoter region of was mapped using truncated 5′ end regions fused to a promoterless green fluorescent protein gene. A 9·5 kb region, adjacent to the sMMO structural gene cluster, was analysed. Downstream (3′) from the last gene of the operon, , four ORFs were found, , , and . shows significant identity to the large subunit of the bacterial chaperonin gene, . In the opposite orientation, two genes, and , showed significant identity to two-component sensor–regulator system genes. Next to , a gene encoding a putative -dependent transcriptional activator, was identified. The and genes were mutated by marker-exchange mutagenesis and the effects of these mutations on the expression of sMMO was investigated. sMMO transcription was impaired in both mutants. These results indicate that and are essential for the expression of sMMO in (Bath).

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2003-07-01
2019-10-18
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