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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 the synthesis of sinomenine by hydroxylation and transformation of sinomenine for the first time, and optimize the conversion conditions to obtain the best process. Methods Ten strains with hydroxylation ability were used to transform sinomenine. TLC and HPLC-MSn methods were used to detect the original drug and its transformants. Highly active strains and optimal conditions for hydroxylation of sinomenine were optimized. . Results After 10 strains were screened, there were 3 strains that could hydroxylate sinomenine. After rescreening, Cunninghamella M2 was the best active strain: under the shaking speed of 180 r/min, the optimal culture time was 6 days, and the initial pH was 6.5. At a culture temperature of 28°C and a maximum substrate feed concentration of not more than 200 mg/L, the hydroxylation rate of sinomenine was greater than 95%. Conclusion Cunninghamella M2 transforms sinomenine to produce its hydroxylated compound with high specificity and high conversion efficiency.