Antibacterial Activity of Recombinant Porcine β-Defensin 2

Received: Revised: Accepted: Published online: September 03, 2018 December 03, 2018 December 12, 2018 March 07, 2019 To express and purify the recombinant porcine β-defensin 2 (rpBD-2) using yeast Pichia Pastoris expression system and in vitro characterization of it’s antibacterial activity. rpBD-2 was expressed and then purified using methylotrophic yeast Pichia Pastoris expression system. To assess its antimicrobial activity against Escherichia coli, Salmonella paratyphi A, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus pumilus, and Bacillus subtilis, liquid growth inhibition assay was used. The antibacterial activity was further kinetically evaluated against representative bacterial strains E. coli and S. aureus. The hemolysis activity of rpBD-2 was also checked against porcine erythrocytes. rpBD2 was expressed and purified successfully using yeast Pichia Pastoris expression system. The yield of final product was 9 mg per liter of the harvested supernatant. rpBD-2 possessed antimicrobial activity against E. coli, S. aureus, S. epidermidis, B. subtilis, and B. pumilus at 33.3-66.6 μM and killed all E. coli and S. aureus within 30 min. Negligible hemolytic activity was observed against porcine erythrocytes. We have successfully developed rpBD-2 as an effective antibacterial peptide against both gram-positive and negative bacteria. rpBD-2 will be a potential novel therapeutic agent against bacterial infection. ©2018 PVJ. All rights reserved