SUMO mediating fusion expression of antimicrobial peptide CM4 from two joined genes in Escherichia coli.
Antibacterial peptide CM4 (ABP-CM4) is a small cationic peptide with broad-spectrum activities against bacteria, fungi, and tumor cells, which may possibly be used as an antimicrobial agent. To improve the expression level of CM4 in Escherichia coli, two tandem repeats of CM4 genes were cloned into the vector pSUMO to construct an expression vector pSUMO-2CM4. The fusion protein SUMO-2CM4, purified by Ni(2+)-chelating chromatography, was cleaved by hydroxylamine hydrochloride to release recombinant CM4. After the cleaved sample was re-applied to a Ni-IDA column, finally, about 48 mg recombinant CM4 was obtained from 1 L bacterial culture with no less than 96% purity, which was the highest yield of CM4 reported so far.
Li
Jian Feng
JF
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing, 210046, Jiangsu, People's Republic of China.
Zhang
Jie
J
Zhang
Zhen
Z
Kang
Chun Tao
CT
Zhang
Shuang Quan
SQ
D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006868:Hydrolysis; D019811:Hydroxylamine / Q000378:metabolism; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D025842:SUMO-1 Protein / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D023181:Antimicrobial Cationic Peptides; C534224:CM4 peptide, Bombyx mori; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; D019811:Hydroxylamine