4-羟基环孢素高产菌株诱变筛选及培养基优化

摘要

An ultraviolet-lithium chloride mutagenesis strategy was carried out on the [(4'-OH)MeLeu] 4-CsA producing strain,Nonomuraea dietziae.The production of mutant strain reached 208 mg/L and increased by 26.8％.Then the fermentation medium was optimized by single-factor and response surface methods.The optimal medium was determined as follows (materials/concentrations in g/L):glycerol/26.5,dextrin/11.6,peptone/9.7,yeast extract powder/3.5,soybean meal/ 15.0,soybean oil/2.0,MgSO4·7H2O/0.40,MnSO4· H2O/0.010,and (NH4)2MoO4/0.010.The final production reached 324 mg/L and increased by 97.6％ compared with the initial value.%以4-羟基环孢素转化菌Nonomuraea dietziae为出发菌,采用紫外-氯化锂复合诱变的方法,筛选得到一株产量达208 mg/L的稳定遗传菌J263,较出发菌164 mg/L的产量提高26.8％.在此基础上,采取单因素法和响应面法针对诱变菌的发酵培养基碳氮源进行系统优化,并对金属离子的添加进行考察.最终获得最佳的培养基配方(甘油26.5 g/L,糊精11.6 g/L,蛋白胨9.7 g/L,酵母浸粉3.5 g/L,大豆饼粉15.0 g/L,大豆油2.0g/L,硫酸镁0.40 g/L,硫酸锰0.010 g/L,钼酸铵0.010g/L).在最优化条件下,4-羟基环孢素产量可达324 mg/L,较出发菌提高97.6％.