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[颗粒状甲烷单加氧酶在不同宿主细胞中的异源表达]

[Heterologous expression of particulate methane monooxygenase in different host cells].

作者信息

Han Bing, Su Tao, Yang Cheng, Jiang Hao, Wu Hao, Zhang Chong, Li Xin, Xing Xinhui

机构信息

Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2009 Aug;25(8):1151-9.

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.

摘要

甲烷营养菌利用甲烷作为唯一的碳源和能源,这导致其生长缓慢、细胞密度低,并阻碍了其工业应用。一种有前景的解决方案是在其他能够易于以高细胞密度培养的宿主细胞中异源表达甲烷单加氧酶(MMO)。我们通过选择不同的启动子和不同的宿主细胞,系统地探索了pMMO功能表达的可能性。结果表明,重组体能够将甲烷氧化为甲醇。特别地,在氧化产物中还能检测到乙醇,但酶活性不稳定,这意味着在宿主细胞中表达的pMMO可能发生了一些变化。此外,SDS-PAGE分析表明,许多重组体能够表达pMMO的亚基,但无法检测到酶活性。总之,pMMO在宿主细胞中的正确折叠对于其功能表达很重要。

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