Open AccessFunctional expression of the particulate methane mono‐oxygenase gene in recombinant Rhodococcus erythropolis
Author(s) -
Gou Zhongxuan,
Xing XinHui,
Luo Mingfang,
Jiang Hao,
Han Bing,
Wu Hao,
Wang Lei,
Zhang Fei
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00363.x
Subject(s) - recombinant dna , shuttle vector , rhodococcus , chemistry , oxygenase , microbiology and biotechnology , gene cluster , gene , gene expression , biochemistry , biology , enzyme , vector (molecular biology)
In order to construct an expression system for the particulate methane mono‐oxygenase (pMMO) gene ( pmo ), the structural gene cluster pmo CAB 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 Cu 2+ and Zn 2+ concentrations on cell growth and pMMO activity in ethane‐containing medium were examined. It was discovered that 7.5 μM Cu 2+ and 1.8 μM Zn 2+ 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.