Heterologous expression of soluble methane monooxygenase genes in methanotrophs containing only particulate methane monooxygenase

被引:35
作者
Lloyd, JS [1 ]
De Marco, P [1 ]
Dalton, H [1 ]
Murrell, JC [1 ]
机构
[1] Univ Warwick, Dept Biol Sci, Coventry CV4 7AL, W Midlands, England
基金
英国自然环境研究理事会; 英国生物技术与生命科学研究理事会;
关键词
heterologous expression; methane oxidation; methane monooxygenase; methanotroph; regulation;
D O I
10.1007/s002030050723
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The methanotrophs Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b contain particulate methane monooxygenase (pMMO) and soluble methane monooxygenase (sMMO) genes. Other methanotrophs such as Methylomicrobium album BG8 and Methylocystis parvus OBBP contain only pMMO genes. Although molecular genetic techniques are poorly developed in methanotrophs, sMMO genes were expressed in methanotrophs normally containing only pMMO genes. This was achieved by conjugation using broad-host-range plasmids containing the native promoter and sMMO genes from Me, capsulatus (Bath) and Ms. trichosporium OB3b, sMMO genes derived from Ms. trichosporium OB3b were expressed in an active form in Mcy. parvus OBBP and in Mm. album BG8. Therefore, all of the genes required for active sMMO synthesis were contained on the broad-host-range plasmids and were expressed in the heterologous hosts. Constitutive synthesis of pMMO was observed in Mm. album BG8 when grown at high and low copper-to-biomass ratios, while transcription of the recombinant sMMO genes was only observed under growth conditions of low copper-to-biomass ratios. Therefore, the regulatory protein(s) for sMMO synthesis was also present on the plasmid used, or the heterologous host contained a regulatory system for sMMO. Expression of sMMO genes in methanotrophs containing only pMMO will assist further investigations on the expression and regulation of MMO genes in methanotrophs.
引用
收藏
页码:364 / 370
页数:7
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