Akademik Veri Yönetim Sistemi
INTERNATIONAL JOURNAL OF PEPTIDE RESEARCH AND THERAPEUTICS, cilt.29, sa.5, ss.1-11, 2023 (SCI-Expanded)
Heterologous expression of the plant-derived snakin-Z (SNK-Z) peptide was carried out in Escherichia coli BL21 and Mach 1 strains using the pET14b, and pGEX-6P-1 vectors, and its tertiary structure was determined. The most efficient production of the recombinant fusion peptide (GST/SNK-Z) in both strains was achieved by induction with 0.1 mM isopropyl β-D-1-thiogalactopyranoside at 32 °C. GST/SNK-Z with 30.14 kDa which yielded 6.5 mg/L protein, was purified by affinity chromatography followed by dialysis. The GST tags were removed using PreScission protease. The IC50 of GST/SNK-Z was calculated as 12.07 µM for Staphylococcus aureus (ATCC 25923) using nonlinear regression analysis. The secondary structure of recombinant SNK-Z consisted of α-helix and coil sites. It’s 3-D model was generated with a confidence score of -0.20 and a template modeling score of 0.69 ± 0.12. The predicted solvent accessibility and B-factor profile were detected. This is the first report of heterologous expression of SNK-Z in E. coli. Our study provides fundamental data for the large-scale production of SNK-Z, which has a high potential for use in the pharmaceutical industry because of its previously reported antimicrobial effects.