Functional expression of the particulate methane mono-oxygenase gene in recombinant Rhodococcus erythropolis.
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.
Gou
Zhongxuan
Z
Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, China.
Xing
Xin-Hui
XH
Luo
Mingfang
M
Jiang
Hao
H
Han
Bing
B
Wu
Hao
H
Wang
Lei
L
Zhang
Fei
F
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
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D010105:Oxygenases; C021555:methane monooxygenase; D008697:Methane