Recombinant expression and coexpression of oyster defensin and proline‐rich peptide in Komagataella phaffii

• Published in: Biotechnology and Applied Biochemistry
• Main Authors: Erdem Büyükkiraz, Mine, Kesmen, Zülal
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 2021
• Subjects: Crassostrea gigas
• Online Access: http://dx.doi.org/10.1002/bab.2262; https://onlinelibrary.wiley.com/doi/pdf/10.1002/bab.2262; https://onlinelibrary.wiley.com/doi/full-xml/10.1002/bab.2262; https://iubmb.onlinelibrary.wiley.com/doi/pdf/10.1002/bab.2262

Abstract Proline‐rich peptide ( Cg Prp) and defensin ( Cg Def), oyster ( Crassostrea gigas )‐originated antimicrobial peptides (AMPs), were produced by the recombinant technique in Komagataella phaffii GS115 cells. For this purpose, the nucleotide sequences encoding the Cg Prp and Cg Def peptides were synthesized by the recursive PCR technique, and ligated in pPICZaA expression vector. Additionally, the expression cassettes of pPICZαA‐ Cg Def and pPICZαA‐ Cg Prp were combined using in vitro multimer ligation strategy to construct the coexpression vector pPICZaA‐ Cg Prp‐ Cg Def. The expression and coexpression vectors transformed into K. phaffii GS115 cells by electroporation. At the end of the 0.5% methanol‐induced expression stage for 96 h, the recombinant peptides were purified from the culture medium. The concentrations of purified peptides were changed between 1.05 and 1.21 mg/L. The recombinant peptides successfully inhibited the growth of tested Gram‐positive bacterial strains belonging to Staphylococcus aureus , methicillin‐resistant S. aureus (MRSA), Listeria monocytogenes , and Bacillus cereus . The minimum inhibitory concentrations (MIC) of recombinant Cg Prp, Cg Def, and Cg Prp‐ Cg Def peptides against tested bacteria were in the range of 12.50–25.00, 18.75–75.00, and 5.80–11.60 pg/μl, respectively. The results of the study proved that the recombinant Cg Prp, Cg Def, and Cg Prp‐ Cg Def peptides expressed in K. phaffii might have good potential for the inhibition of common Gram‐positive pathogenic bacteria, including drug‐resistant MRSA.