Expression and purification of the antimicrobial peptide cecropin AD by fusion with cationic elastin-like polypeptides.

Ke Yang, Yujie Su, Junhua Li, Jun Sun, Yanjun Yang

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, PR China.

Protein expression and purification 2012 Oct

filter terms:

Cationic elastin-like polypeptides (CELP) are thermally responsive polypeptides that undergo an inverse temperature phase transition, and the recombinant CELP fusion proteins may be purified by inverse transition cycling (ITC). To obtain high-purity antimicrobial peptide cecropin AD (CAD), CELP was placed at the N-terminus of CAD and the expression vector pET28a-CELP-CAD was constructed. The expression vector was then transformed into Escherichia coli BL21 (DE3) to express the recombinant protein. After three rounds of ITC, enterokinase digestion and another hot spin, 1.2mg recombinant CAD was purified from 100ml culture medium. The antimicrobial test indicated that the high-purity CAD had strong antimicrobial activity against E. coli and Staphylococcus aureus. Copyright © 2012. Published by Elsevier Inc.

Citation

Ke Yang, Yujie Su, Junhua Li, Jun Sun, Yanjun Yang. Expression and purification of the antimicrobial peptide cecropin AD by fusion with cationic elastin-like polypeptides. Protein expression and purification. 2012 Oct;85(2):200-3