title: Production of a cytotoxic cationic antibacterial peptide in Escherichia coli using SUMO fusion partner.
abstract: 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. We report here the application of small ubiquitin-related modifier (SUMO) fusion technology to the expression and purification of cationic antibacterial peptide ABP-CM4. The fusion protein expressed in a soluble form was purified to a purity of 90% by Ni-IDA chromatography and 112 mg protein of interest was obtained per liter of fermentation culture. After the SUMO-CM4 fusion protein was cleaved by the SUMO protease at 30 degrees C for 1 h, the cleaved sample was re-applied to a Ni-IDA. Finally, about 24 mg recombinant CM4 was obtained from 1 l fermentation culture with no less than 96% purity and the recombinant CM4 had similar antimicrobial properties to the synthetic CM4. Thus, the SUMO-mediated peptide expression and purification system potentially could be employed for the production of recombinant cytotoxic peptides.
authors: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing, 210046, People's Republic of China.
authors: Jian Feng
authors: JF
authors: Li
authors: Jie
authors: J
authors: Zhang
authors: Ren
authors: R
authors: Song
authors: Jia Xin
authors: JX
authors: Zhang
authors: Yang
authors: Y
authors: Shen
authors: Shuang Quan
authors: SQ
authors: Zhang
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism* / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005285:Fermentation; D005798:Genes, Bacterial; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics*
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; C534224:CM4 peptide, Bombyx mori; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D007544:Isopropyl Thiogalactoside