黄素单加氧酶在大肠杆菌中的表达及靛蓝的生物合成

Study on Cloning and Expression of Flavin-containing Monooxygenase in Escherichia Coli and Indigo Biosynthesis

为实现生物转化法合成靛蓝,构建了携带黄素单加氧酶基因的工程菌E.coli BL21(pET28a/fmo)。工程菌经诱导表达后,加入吲哚进行转化,薄层分析确定产物即靛蓝。文章也对培养基中底物浓度、诱导剂、转化温度、转化时间等影响靛蓝合成的因子进行了初步探讨。结果表明,在合适的条件下,工程菌转化吲哚合成靛蓝产物浓度最高达到48.28mg/L,转化率到达64.4%。

In order to produce indigo through the biosynthesis method, an engineered E.coli carrying gene fmo named E.coli BL21（p ET28a/fmo） was constructed. Cultivated and induced in LB medium supplemented with indole,the engineered E.coli can transform the indole into indigo which can be identified by thin layer chromatography. The paper also researches the optimums conditions of biosynthesis. The results show that the 48.28mg/L of indigo was harvested and the conversion rate was as much as 64.4%.