Expression and purification the antimicrobial peptide CM4 in Escherichia coli.
The antimicrobial peptide CM4 is a 35-residue cationic peptide. To explore a new approach for the expression and purification of CM4 in Escherichia coli, the CM4 gene was cloned into the vector pET32a to construct an expression vector pET32a-CM4. The fusion protein Trx-CM4, purified by Ni(2+)-chelating chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant CM4. Purification of recombinant CM4 was achieved by reverse HPLC chromatography, and about 1.4 mg/l active recombinant CM4 with the purity more than 98% was obtained. The recombinant CM4 showed antimicrobial activities that were similar to synthetic one.
Zhou
Liangfan
L
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing, 210046, China.
Lin
Qingping
Q
Li
Baocun
B
Li
Nannan
N
Zhang
Shuangquan
S
D000890:Anti-Infective Agents / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics* / Q000494:pharmacology; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005658:Fungi / Q000187:drug effects; D005822:Genetic Vectors; D006090:Gram-Negative Bacteria / Q000187:drug effects; D019811:Hydroxylamine / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; C534224:CM4 peptide, Bombyx mori; D011993:Recombinant Fusion Proteins; D019811:Hydroxylamine