Abstract
Antimicrobial peptides are one of the most promising peptide-based drugs due to their enormous potential as novel biopharmaceuticals in both human and animal industries. In order to develop strategies to over produce such molecules, heterologous production of a modified version of clavanin A, here named clavanin MO (clavMO), was successfully achieved in the methylothopic yeast Pichia pastoris. ClavMO was fused to thioredoxin as a carrier protein and the construction was tested using two promoters, PAOX1 and PGAP, based on either induced or constitutive expression systems, respectively. After growth in 5 L Bioreactor, clavMO-thio was recovered and purified through size exclusion chromatography. Our findings show that both constitutive and inducible expression systems produce active clavMO fused to thioredoxin against both Gram-negative Klebsiella pneumoniae and Gram-positive Staphylococcus aureus microorganisms.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-015-0129-0) contains supplementary material, which is available to authorized users.
Highlights
Antimicrobial peptides (AMPs) are one of the most promising peptide-based drugs due to their enormous potential as novel biopharmaceutical compounds for the human and animal health industries as well as for their application in agriculture (Agyei and Danquah 2011; Mulder et al 2013a; Silva et al 2011a)
Our findings show that the induced system produces active clavanin MO (clavMO) against the microorganisms K. pneumoniae and S. aureus
Production of clavMO X-33/pPICZαA-clavMO and X-33/pGAPZαB-clavMO were selected for growth experiments and heterologous peptide production
Summary
Antimicrobial peptides (AMPs) are one of the most promising peptide-based drugs due to their enormous potential as novel biopharmaceutical compounds for the human and animal health industries as well as for their application in agriculture (Agyei and Danquah 2011; Mulder et al 2013a; Silva et al 2011a). The production of AMPs using heterologous systems has many advantages, such as allowing post-translational modification and permitting researchers to develop the best genetic strategy to increase its production, and most importantly, there is an extensive flexibility in microbial systems to be modified and scaleup (Mulder et al 2013b). The heterologous expression of AMPs has been reported to be successfully performed in diverse organisms such as bacteria, plants and yeast (Parachin et al 2012).
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