Heterologous expression of particulate methane monooxygenase in different host cells.
- Author:
Bing HAN
1
;
Tao SU
;
Cheng YANG
;
Hao JIANG
;
Hao WU
;
Chong ZHANG
;
Xin LI
;
Xinhui XING
Author Information
1. Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Gene Expression Regulation, Enzymologic;
Genetic Vectors;
genetics;
Methane;
metabolism;
Methanococcaceae;
enzymology;
Methanol;
metabolism;
Oxygenases;
biosynthesis;
genetics;
Promoter Regions, Genetic;
Protein Folding;
Recombinant Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2009;25(8):1151-1159
- CountryChina
- Language:Chinese
-
Abstract:
Methanotrophs use methane as the sole carbon and energy source, which cause slow growth, low cell density and hinder its industrial applications. One promising solution is to heterologously express methane monooxygenase (MMO) in other host cells that can be easily cultivated at high cell density. We systematically exploited the possibility of functional expression of pMMO by choosing different promoters and different host cells. The results showed that the recombinants could oxidize methane to methanol. In particular, ethanol could also be detected in the oxidized products, but the enzyme activity was instable, implying that some changes of pMMO expressed in the host cells might have occurred. In addition, SDS-PAGE analysis showed that many recombinants could express the subunits of pMMO, but the enzyme activity could not be detected. In conclusion, correct fold of pMMO in the host cells is important for its functional expression.
