title: Expression and one-step purification of the antimicrobial peptide cathelicidin-BF using the intein system in Bacillus subtilis.
abstract: The intein expression system has been widely applied in Escherichia coli to express various proteins and peptides. However, the removal of endotoxin from the recombinant proteins expressed in E. coli is very difficult and therefore complicates the purification process. In this study, we constructed an intein-based expression vector for an antimicrobial peptide (cathelicidin from Bungarus fasciatus) and expressed the intein fusion peptide in a Bacillus subtilis expression system. The fusion peptide was secreted into the culture medium, identified by Western blot and purified by affinity chromatography and intein self-cleavage in just one step. Approximately, 0.5 mg peptide was obtained from 1 litre of culture medium. The purified peptide showed antimicrobial activity. Our results indicate that the intein expression system may be a safe and efficient method to produce soluble peptides and proteins in B. subtilis.
authors.affiliation: Key University Laboratory of Biotechnology and Utilization of Bio-resource of Shandong, College of Life Science, Dezhou University, 566 University Road West, Dezhou, 253023, Shandong Province, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: He
authors.forename: Ai-yun
authors.initials: AY
authors.lastname: Fu
authors.forename: Tian-jiao
authors.initials: TJ
authors.lastname: Li
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D017838:Bungarus; D054804:Cathelicidins / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D002846:Chromatography, Affinity; D003470:Culture Media / Q000737:chemistry; D047668:Inteins / Q000502:physiology*; D008826:Microbial Sensitivity Tests; D019154:Protein Splicing; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012995:Solubility
chemical_list: D023181:Antimicrobial Cationic Peptides; D054804:Cathelicidins; D003470:Culture Media; D011993:Recombinant Fusion Proteins