(Bath) uses a soluble methane monooxygenase (sMMO) to catalyse the oxidation of methane to methanol. sMMO is comprised of three components; A, B and C. Protein C (the reductase) transfers electrons from NADH to protein A (the hydroxylase) which contains the active site, and protein B regulates this electron flow. The five genes encoding the sMMO proteins and their subunits are clustered and have been cloned in . A DNA fragment containing , the gene encoding protein B, was subcloned into pT7–5, a plasmid of the T7 RNA polymerase promoter expression system. Upon induction, expressed protein B which was fully functional after purification. The gene encoding protein C, , was amplified with unique restriction sites at each end using the polymerase chain reaction and then subcloned into pT7–7 (a plasmid similar to pT7–5 but containing its own ribosome-binding site and ATG start codon). Protein C expressed in was also found to be functional. This is the first report of the functional expression of methanotroph methane monooxygenase genes in a heterologous host and represents a significant step forward in our analysis of the assembly and catalysis of sMMO.


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