Expression and purification of an antimicrobial peptide by fusion with elastin-like polypeptides in Escherichia coli.
Different carrier molecules have been fused to antimicrobial polypeptides (AMPs) to facilitate recombinant protein expression and purification. Some of them have improved the stability of AMPs and reduced the toxicity to host cells, but most current strategies still have some problems to be solved such as poor yield, low purity, high expense, time-consumption, and difficulty in scaling-up. Here, we introduced the elastin-like polypeptides (ELPs) as a fusion partner to express an antimicrobial polypeptide halocidin18 (Hal18). By the reversible soluble-insoluble phase transition, 69 mg of the fusion protein were purified from 1 l of culture medium with the purity of nearly 95%. After cleavage with hydroxylamine, the ELP's tag was easily separated from Hal18 in the next round of inverse transition cycle and Hal18 (1.7 mg, approximately 1.9 kDa) was mainly found in the supernatant with a recovery of about 47% and purity of 60%. Antimicrobial activity showed that Hal18 had strong antimicrobial activity against Escherichia coli and Micrococcus luteus but weak activity against Pichia pastoris.
Hu
Fan
F
College of Life Science, Hubei University, Wuhan, 430062, China.
Ke
Tao
T
Li
Xin
X
Mao
Pei Hong
PH
Jin
Xiang
X
Hui
Feng Li
FL
Ma
Xiang Dong
XD
Ma
Li Xin
LX
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D004549:Elastin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology
D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins; D004549:Elastin