title: Functional expression of the particulate methane mono-oxygenase gene in recombinant Rhodococcus erythropolis.
abstract: In order to construct an expression system for the particulate methane mono-oxygenase (pMMO) gene (pmo), the structural gene cluster pmoCAB amplified from Methylosinus trichosporium OB3b was inserted into a shuttle vector pBS305 under the control of a dsz promoter and transformed into Rhodococcus erythropolis LSSE8-1. A stable transformant was successfully obtained using ethane as the sole carbon source. Fluorescence in situ hybridization results showed that the dsz promoter allowed the pmo genes to be transcribed in the recombinant strain. The effects of Cu2+ and Zn2+ concentrations on cell growth and pMMO activity in ethane-containing medium were examined. It was discovered that 7.5 microM Cu2+ and 1.8 microM Zn2+ were suitable to achieve high cell concentration and pMMO activity, but the amount of methanol accumulated during methane oxidation by the recombinant strain was still low.
authors.affiliation: Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, China.
authors.forename: Zhongxuan
authors.initials: Z
authors.lastname: Gou
authors.forename: Xin-Hui
authors.initials: XH
authors.lastname: Xing
authors.forename: Mingfang
authors.initials: M
authors.lastname: Luo
authors.forename: Hao
authors.initials: H
authors.lastname: Jiang
authors.forename: Bing
authors.initials: B
authors.lastname: Han
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Fei
authors.initials: F
authors.lastname: Zhang
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008697:Methane / Q000378:metabolism*; D020581:Methylosinus / Q000201:enzymology / Q000235:genetics*; D005810:Multigene Family; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D004274:DNA, Recombinant; D010105:Oxygenases; C021555:methane monooxygenase; D008697:Methane