重组阿片肽包涵体的溶解工艺

Solubilization of the Recombinant Casomorphin Inclusion Body

通过SDS PAGE凝胶电泳确定了目的重组阿片肽存在于包涵体中,系统研究了影响大肠杆菌表达的重组阿片肽包涵体溶解的因素,确定了重组阿片肽包涵体制备溶解的最佳工艺条件,即菌体超声破碎5min,间隔时间6s,破碎效果最好;包涵体洗涤温度为4℃,尿素洗涤液的浓度为2mol/L,洗涤时间7～8h,8mol/L的尿素溶液对包涵体进行溶解,蛋白质质量浓度约为600μg/mL,其效果最佳.

The target recombinant casomorphin was found in inclusion body in E.coli by SDSPAGE. The technique for solubilization of the recombinant casomorphin inclusion body was studied. The optimal solubilization conditions were obtained: E.coli cells were disrupted by ultrasonic in 5 minutes, alternation time was 6 s; the inclusion body was washed at 4 ℃,the washing concentration of urea was 2.0 mol/L, and the washing time was 7～8 hours. The optimal concentration of urea was 8 mol/L. After solubilizing,the protein concentration was about 600 μg/mL.