Production of a polar fish antimicrobial peptide in Escherichia coli using an ELP-intein tag
| Autor: | Kethly S. Nobre, Daniel Amaro Sousa, Kelly C.L. Mulder, Nádia Skorupa Parachin, Octavio L. Franco |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Peptide Biosynthesis
0106 biological sciences 0301 basic medicine Recombinant Fusion Proteins medicine.medical_treatment Antimicrobial peptides Bioengineering Peptide Biology medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Chemical synthesis Inteins 03 medical and health sciences 010608 biotechnology Escherichia coli medicine chemistry.chemical_classification Protease General Medicine Antimicrobial Anti-Bacterial Agents Elastin Chemically defined medium 030104 developmental biology chemistry Biochemistry Fermentation Peptides Intein Genome Bacterial Biotechnology |
| Zdroj: | Journal of Biotechnology. 234:83-89 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2016.07.021 |
| Popis: | An important aspect related to infectious pathogens is their exceptional adaptability in developing resistance, which leads to a perpetual challenge in the discovery of antimicrobial drugs with novel mechanisms of action. Among them, antimicrobial peptides (AMPs) stand out as promising anti-infective molecules. In order to overcome the high costs associated with isolation from natural sources or chemical synthesis of AMPs we propose the expression of Pa-MAP 2, a polyalanine AMP. Pa-MAP 2 was fused to an ELP-intein tag where the ELP (Elastin-like polypeptide) was used to promote aggregation and fast and cost-effective isolation after expression, and the intein was used to stimulate a controlled AMP release. For these, the vector pET21a was used to produce Pa-MAP 2 fused to the N-termini region of a modified Mxe GyrA intein followed by 60 repetitions of ELP. Purified Pa-MAP 2 showed a MIC of 25μM against E. coli ATCC 8739. Batch fermentation demonstrated that Pa-MAP-2 can be produced in both rich and defined media at yields 50-fold higher than reported for other AMPs produced by the ELP-intein system, and in comparable yields to expression systems with protease or chemical cleavage. |
| Databáze: | OpenAIRE |
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