Abstract
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.
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21 August 2023
A Correction to this paper has been published: https://doi.org/10.1007/s10989-023-10560-z
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Acknowledgements
The Council of Higher Education (CoHE) is acknowledged for granting of T. Teker’s doctoral study in the frame of 100/2000 CoHE Doctoral Scholarship Program. The Scientific and Technological Research Council of Turkey is gratefully acknowledged by T. Teker for granting a fellowship within the scope of the BIDEB 2211-A National PhD Scholarship Program. We extend our thanks to G.B. Albayrak -bilingual-proficient- for grammatically correcting this article.
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This study was funded by Scientific Research Projects Coordination Unit of Istanbul University. Project number:36971.
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Conceptualization: [T. Teker, G. Albayrak, K. Turan]; Methodology: [T. Teker, G. Albayrak, K. Turan]; Formal analysis and investigation: [T. Teker, G. Albayrak, K. Turan]; Writing - original draft preparation: [T. Teker]; Writing - review and editing: [G. Albayrak, K. Turan]; Funding acquisition: [G. Albayrak]; Supervision: [G. Albayrak, K. Turan]. All authors read and approved the final manuscript.
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Teker, T., Albayrak, G. & Turan, K. Heterologous Expression and Initial In Silico Characterization of a Novel Snakin-Z Peptide. Int J Pept Res Ther 29, 84 (2023). https://doi.org/10.1007/s10989-023-10556-9
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DOI: https://doi.org/10.1007/s10989-023-10556-9