1. Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG
- Author
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Xiang Liu, Yining Ding, Yuhan Shen, Sizhuo Liu, Yuehua Liu, Yuting Wang, Shikun Wang, Claudio Orlando Gualerzi, Attilio Fabbretti, Lili Guan, Lingcong Kong, Haipeng Zhang, Hongxia Ma, and Chengguang He
- Subjects
antimicrobial peptides ,prokaryotic expression ,antibacterial mechanism ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
The antimicrobial peptide LRGG (LLRLLRRGGRRLLRLL-NH2) was designed and chemically synthesized in a study conducted by Jia et al. Gram-negative bacteria were found to be sensitive to LRGG and exhibited a high therapeutic index. Genetic engineering methods were used to create the prokaryotic fusion expression vector pQE-GFP-LRGG, and the resulting corresponding fusion protein GFP-LRGG was subsequently expressed and purified. The precursor GFP was then removed by TEV proteolysis, and pure LRGG was obtained after another round of purification and endotoxin removal. The prokaryotic-expressed antimicrobial peptide LRGG displays a broad-spectrum antibacterial effect on Gram-negative bacteria, and its minimum inhibitory activity (MIC) against Escherichia coli can reach 2 μg/mL. Compared to the chemically synthesized LRGG, the prokaryotic-expressed LRGG exhibits similar temperature, pH, salt ion, serum stability, and cell selectivity. Furthermore, prokaryotic-expressed LRGG showed excellent therapeutic effects in both the infection model of cell selectivity and no embryotoxicity in a Galleria mellonella infection model. The mechanism by which LRGG causes bacterial death was found to be the disruption of the Gram-negative cell membrane.
- Published
- 2024
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