Abstract
Melittin (MLT) is a small cationic peptide discovered from the bee venom. It is used as an antimicrobial agent due to its broad-spectrum activities against bacteria, fungi and tumor cells. But the sources limit its applications. Therefore, the small ubiquitin-related modifier (SUMO) fusion technology was reported for high-level expression of Melittin. pET-3c-SUMO-Melittin plasmid was constructed, and the fusion protein (SUMO-Melittin) was expressed in a soluble form and purity by Ni2+-NTA chromatography. After the SUMO-Melittin fusion protein was cleaved by the SUMO protease, the cleaved sample was purified again by a Ni2+-NTA. Finally, about 25 mg recombinant Melittin was obtained from 1L fermentation culture with more than 95% purity. The recombinant Melittin exhibited similar cytotoxicity and antimicrobial properties as the synthetic Melittin. Thus, a system for high-level expression of Melittin was successfully established in this study and could be used for preparation of similar antimicrobial peptides.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (51973081); Jilin Collaborative Innovation Center for Antibody Engineering (20180623045TC); Project Agreement for Science & Technology Development, Jilin Province (20170414022GH); The Education Department of Jilin province “13th Five Year Plan” science and technology research projects (JJKH20180829KJ).
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Chen, Qc., Liu, L., Yu, TY. et al. High-Level Expression and Purification of Melittin in Escherichia coli Using SUMO Fusion Partner. Int J Pept Res Ther 27, 9–15 (2021). https://doi.org/10.1007/s10989-020-10060-4
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DOI: https://doi.org/10.1007/s10989-020-10060-4