Expression and one-step purification of the antimicrobial peptide cathelicidin-BF using the intein system in Bacillus subtilis.
Qing He, Ai-yun Fu, Tian-jiao Li
Author Information
Qing He: 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.
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.
References
Appl Environ Microbiol. 2008 Feb;74(4):1039-49
[PMID: 18156317]
Protein Expr Purif. 2013 May;89(1):16-24
[PMID: 23422783]
