Production and characterization of therapeutic peptides with potential antimicrobial properties.
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Date
2024
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Publisher
Univeristy of the Western Cape
Abstract
Bacterial drug resistance is a current and growing serious health problem worldwide because of the long-term and negligent use of antibiotics. Finding new antibiotics and shepherding them through the clinical trials and approval process is becoming increasingly difficult. In addition, the declining number of new innovative antibiotic candidates has worsened this burden, necessitating the development of new classes of antimicrobial agents. Antimicrobial peptides (AMPs) are protein molecules that often form part of the innate immune system and cellular defense mechanisms found in many organisms. AMPs are promising antibiotic alternatives for the treatment of drug-resistant infections. They are, however, constrained by their high manufacturing costs. Genetic engineering and heterologous recombinant peptide expression are promising approaches for producing such molecules at a low cost. In this study, peptide Gene 1 and peptide Gene 2 obtained from the bioinformatics analysis on marine metagenomes sampled from the TARA Oceans Project were produced using the heterologous recombinant peptide expression method. The peptide Gene 1 and peptide Gene 2 were cloned into the pPIC9 vector, transformed, and integrated into the P. pastoris genome. The expression was observed in BMMY medium, pH 6, 1% methanol, and 48 hr, 72 hr, 96 hr with a band at a molecular mass of 6.05 kDa for peptide Gene 1 and 11.37 kDa for peptide Gene 2 following SDS-PAGE analysis.
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Keywords
Antimicrobial Activity, Antimicrobial Peptides, Antimicrobial Resistance, Classification, Cloning