%0 Journal Article
%T Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG.
%A Liu X
%A Ding Y
%A Shen Y
%A Liu S
%A Liu Y
%A Wang Y
%A Wang S
%A Gualerzi CO
%A Fabbretti A
%A Guan L
%A Kong L
%A Zhang H
%A Ma H
%A He C
%J Int J Mol Sci
%V 25
%N 13
%D 2024 Jun 27
%M 39000180
%F 6.208
%R 10.3390/ijms25137072
%X 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.