微生物转化合成羟基化青藤碱及其条件优化

Synthesis of hydroxy-sinomenine by microbial transformation and optimization of its transformation conditions

目的首次对青藤碱进行微生物羟基化转化合成研究,并对转化条件进行优化得到最佳工艺。方法选取10株具有羟基化能力的菌株对青藤碱进行转化,采用TLC、HPLC-MSn方法检测原药及其转化物,优选出能使青藤碱羟基化的高活性菌株和最佳转化条件。结果10株菌株经过筛选,有三株菌可使青藤碱羟基化,经过复筛Cunninghamella M2为最佳活性菌株:在摇床转速180r/min下,最适培养时间为6d,初始pH为6.5,培养温度为28℃,最大底物投料浓度不大于200mg/L时,可使青藤碱羟基化率大于95%。结论Cunninghamella M2转化修饰青藤碱生成其羟基化物的专属性强、转化效率高。

Objective To study on the synthesis and transformation of sinomenine microbial hydroxylation for the first time, and obtain the best technique on the basis of the optimization of transformation conditions. Methods Ten microbe strains with hydroxylation abilities were used in the transformation of sinomenine. The methods of TLC and HPLC-MSn were applied to identify the original sinomenine and its metabolites, and to optimize the microbe strains with high activities and the best transformation condition for the hydroxylation of sinomenine. Results After the screening there were three microbe strains showed the hydroxylation abilities among ten, and CunninghameUa M2 was the best one after secondary screening. When the rotation speed was 180 r/min, optimum culture time was 6 days, initial pH value was 6.5, culture temperature was 28℃, and highest concentration of feeding substrate was below 200 mg/L, the hydroxylation rate of sinomenine would be over 95%. Conclusion Cunninghamella M2 can transform and modify sinomenine into hydroxy-sinomenine, which has strong specificity and high transformation efficiency.