Study on expression,puriifcation and properties of recombinant human Chymotrypsin / 中国生化药物杂志

ABSTRACT

Objective To study the prokaryotic expression, puriifcation and properties of recombinant human Chymotrypsin. Methods The protein was highly expressed in E.coliBL 21 (DE 3) as inclusion body. After refolding and activated with trypsin, the activated protein was obtained and purified with ion-exchange chromatography(CM-FF), some properties of the recombinant human chymotrypsin was investigated. Results The molecular weight of chymotrypsinogen was about 30 KD in SDS-PAGE, total activity recovery rate of CM-FF puriifcation was 93.7%. The recombinant chymotrypsin kept stable from pH 3 to pH 5, and owned good temperature stability. Km was 0.067 mmol/L with BTEE as a substrate. The UV maximum absorption wavelength was 281 nm.Conculsion The recombinant human enzyme was expressed successfully, and a feasible production method to get a high activity of the recombinant human chymotrypsin was established.