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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。
OBJECTIVE: To study the fermentation medium and substrate transformation conditions of a strain of Streptomyces roseochromogenes to improve the conversion of 16α-hydroxyprednisolone. Methods: The target strain TS-58 was obtained by ultraviolet and lithium chloride mutagenesis. The fermentation conditions of shake flask were studied by orthogonal experiment and the effects of different carbon and nitrogen sources on the growth of Streptomyces roseicola were studied. The effects of different substrate concentrations, substrate addition time, liquid loading, metal ions and the addition of co-solvents on the ability to convert 16α-hydroxy prednisolone were investigated. Results: The optimal culture medium was glucose 10 g / L, soluble starch 50 g / L, peptone 10 g / L, soybean cake 25 g / L, potassium dihydrogen phosphate 0.2 g / L zinc sulfate 0.5 g / L. Under the optimum conditions of adding 0.8 mg / ml PEG cosolvent with 5 mg / mL substrate, the conversion of 16α-hydroxyprednisolone reached 13.8%. CONCLUSION: The mutant strain Streptomyces roseochromogenes TS-58 can effectively introduce 16α-hydroxy hydroxyl on prednisolone, which lays the foundation for the industrial production of 16α-hydroxy prednisolone.