生物转化法合成16α-羟基泼尼松龙的发酵工艺研究

目的:研究1株玫瑰产色链霉菌(Streptomyces roseochromogenes)的发酵培养基和底物转化条件,以提高16α-羟基泼尼松龙的转化率。方法:采用紫外与氯化锂复合诱变获得目的菌株TS-58,利用正交实验等方法考查摇瓶发酵条件,研究不同浓度的碳源、氮源对玫瑰产色链霉菌生长的影响,以及不同底物浓度、底物加入时间、装液量、金属离子和添加助溶剂等条件对转化生成16α-羟基泼尼松龙能力的影响。结果:获得最佳转化培养基为葡萄糖10 g/L、可溶性淀粉50 g/L、蛋白胨10 g/L、黄豆饼粉25 g/L、磷酸二氢钾0.2 g/L、硫酸镁0.5 g/L硫酸锌0.5 g/L。在底物投料量5 mg/mL添加0.8 mg/ml PEG助溶剂的最优条件下,16α-羟基泼尼松龙的转化率达到了13.8%。结论:突变株Streptomyces roseochromogenes TS-58能有效地在泼尼松龙上引入16α-羟基羟基,为工业生产16α-羟基泼尼松龙奠定了基础。

Transformation Condition Optimization of 16α-hydroxylation of Prednisolone by Streptomyces roseochromogenes

Objective: The optimum fermentation culture and transformation conditions of mutant strain Streptomyces roseochromogenes TS-58,obtained by UV-LiCl complex mutation,were studied to improve the efficiency of prednisolone 16α-hydroxylation.Method: The optimal fermentation culture and was determined by orthogonal test.The different reaction parameters for maximum conversion of prednisolone were optimized.Result: The optimal fermentation culture contained: glucose 10 g/L,soluble starch 50 g/L,peptone 10 g/L,soybean meal 25 g/L,potassium dihydrogenphosphate 0.2 g/L,magnesium sulfate 0.5 g/L,zinc sulfate 0.5 g/L.While conversion rate of prednisolone to 16α-hydroxyprednisolone arrived at 13.8% under the optimal culture conditions when substrate concentration was 5 mg/ ml and 8 mg/ mL PEG added.Conclusion: The mutant strain Streptomyces roseochromogenes TS-58 can introduce 16α-hydroxyl to prednisolone effectively,which paves the way for the industrial production of 16α-hydroxyprednisolone.