Expression and purification of the antimicrobial peptide cecropin AD by fusion with cationic elastin-like polypeptides.
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.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, PR China.
Ke
K
Yang
Yujie
Y
Su
Junhua
J
Li
Jun
J
Sun
Yanjun
Y
Yang
D000595:Amino Acid Sequence; D001483:Base Sequence; D054807:Cecropins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D003001:Cloning, Molecular; D004549:Elastin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000187:drug effects
D054807:Cecropins; D011993:Recombinant Fusion Proteins; D004549:Elastin