大肠杆菌异源表达pMMO活性中心pmoB及生物催化研究

颗粒甲烷单加氧酶（pMMO）是甲烷氧化菌中催化甲烷氧化生成甲醇的一种酶．Methylococcus capsulatus IMV 3021的pMMO活性位点是pmoB亚基，该亚基是一种可溶性蛋白．我们研究将pmoB亚基进行异源表达及生物催化活性的验证．当培养基中烟酰胺腺嘌呤二核苷酸（NADH）浓度为5 mmol／L时，可以观察到异源表达pmoB亚基具有催化甲烷氧化成甲醇活性，生成甲醇浓度为1．04 mmol／L．研究pMMO活性对于开发能直接将甲烷转化成甲醇的新型、环保催化剂有非常重要意义．

Production and biocatalyst of pmoB subunit from particulate methane monooxygenase in Escherichia coli by heterologous expression

Particulate methane monooxygenase (pMMO) are enzymes that catalyze the oxidation of methane to methanol in the methanotrophs. To localize the active site within the pMMO structure, constructs were generated corresponding to the soluble cupredoxin domains of Methylococcus capsulatus (Bath) pmoB. In this chapter, we describe protocols to express and biocatalyst of the pmoB subunit. The NADH concentration in the growth medium was found to be 5 mM. We show that pmoB subunit by heterologous expression catalyze the oxygenation of methane to methanol. The methanol concentration was 1.04 mmol/L. Understanding pMMO function has important implications for the development of new, environmentally friendly catalysts for the direct conversion of methane to methanol.