基因工程菌SIPI-A0707代谢产物表柔红霉素的分离纯化工艺研究

Study on the separation and purification of 4'-epi-daunorubicin produced by gene engineering strain SIPI-A0707

目的建立天蓝淡红链霉菌基因工程菌(SIPI-A0707)发酵液中表柔红霉素的分离纯化工艺。方法调节发酵液pH值为5,用甲醇抽提表柔红霉素;先采用大孔弱酸性阳离子交换树脂D152,吸附流速为2mL/min,0.05mol/L HCl-50%丙酮解吸;然后采用氯仿-水萃取体系,萃取pH为6.5,反萃取使用pH为1.5的蒸馏水,最后采用大孔吸附树脂Microsphere-1;吸附pH为7,吸附流速3mL/min,50%甲醇解吸分步收集,浓缩冻干。结果表柔红霉素HPLC纯度达到98.1%,总收率为40%以上。结论此工艺在生产上可行性较高,适合工业大量制备。

Objective To establish a separation and purification process of 4＇-epi-daunorubicin from fermentation broth of gene engineering strain Streptomyces coeruleorubidus （SIPI-A0707）. Methods Adjusted the pH value of fermentation broth to 5 and extracted 4＇-epi-daunorubicin with methanol. Firstly macroporous weak acid cation exchange resin D152 was used and the adsorption flow rate was 2mL/min, 0.05mol/L HC1-50% acetone was used as desorption agent. Then chloroform-water extraction system was studied which showed the optimized extraction pH value was 6.5 and distilled water with pH 1.5 was used as reextraction agent. Finally macroporous resin Microsphere-1 was used and the adsorption pH value was 7, the adsorption flow rate was 3mL/min, 50% methanol was used as desorption agent and fractions were collected, decompressed and freeze-dried. Results The HPLC purity of 4＇-epi-daunorubicin was over 98.1%, total yield was over 40%. Conclusion The extraction process of 4＇-epi- daunorubicin is easy to scale-up for industrial preparation.