Expression and purification of recombinant human cytochrome C in Escherichia coli.
- Author:
Huawei CAI
1
;
Lin WAN
;
Hao YANG
;
Lihong CHEN
;
Xiaofeng LU
Author Information
1. Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Cytochromes c;
biosynthesis;
genetics;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
Humans;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification
- From:
Journal of Biomedical Engineering
2007;24(3):620-625
- CountryChina
- Language:Chinese
-
Abstract:
Cytochrome C plays important roles in electron transferring, oxidative stress and apoptosis. In this study, soluble cytochrome C was accumulated in cytoplasm of E. coli by utilizing the co-expression of human cytochrome c and yeast heme lyase from a single plasmid. After ultrasonic disruption of the bacteria, a lot of contaminated proteins were discarded by addition of 350 g/L ammonium sulfate into the supernatant. Then the recombinant human cytochrome C was purified to 80% homogeneity after two times cation exchange chromatography on SP-Sepharose Fast Flow. Yields of cytochrome C greater than 10 mg per liter culture were attained. This efficient system for producing human cytochrome C is helpful for us to understand the roles of this protein in biological processes and therapy of human diseases relevant to apoptosis and oxidative stress.
