Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG

Xiang Liu; YiNing Ding; YuHan Shen; SiZhuo Liu; YuHua Liu; YuTing Wang; ShiKun Wang; Claudio Orlando Gualerzi; Attilio Fabbretti; LiLi Guan; LingCong Kong; HaiPeng Zhang; HongXia Ma; ChengGuang He

doi:10.20944/preprints202405.1656.v1

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preprints.org > biology and life sciences > biochemistry and molecular biology > doi: 10.20944/preprints202405.1656.v1

Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG

Claudio Orlando Gualerzi

Version 1 : Received: 24 May 2024 / Approved: 24 May 2024 / Online: 24 May 2024 (15:39:30 CEST)

How to cite: Liu, X.; Ding, Y.; Shen, Y.; Liu, S.; Liu, Y.; Wang, Y.; Wang, S.; Gualerzi, C. O.; Fabbretti, A.; Guan, L.; Kong, L.; Zhang, H.; Ma, H.; He, C. Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG. Preprints 2024, 2024051656. https://doi.org/10.20944/preprints202405.1656.v1 Liu, X.; Ding, Y.; Shen, Y.; Liu, S.; Liu, Y.; Wang, Y.; Wang, S.; Gualerzi, C. O.; Fabbretti, A.; Guan, L.; Kong, L.; Zhang, H.; Ma, H.; He, C. Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG. Preprints 2024, 2024051656. https://doi.org/10.20944/preprints202405.1656.v1

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MDPI and ACS Style

Liu, X.; Ding, Y.; Shen, Y.; Liu, S.; Liu, Y.; Wang, Y.; Wang, S.; Gualerzi, C. O.; Fabbretti, A.; Guan, L.; Kong, L.; Zhang, H.; Ma, H.; He, C. Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG. Preprints 2024, 2024051656. https://doi.org/10.20944/preprints202405.1656.v1

APA Style

Liu, X., Ding, Y., Shen, Y., Liu, S., Liu, Y., Wang, Y., Wang, S., Gualerzi, C. O., Fabbretti, A., Guan, L., Kong, L., Zhang, H., Ma, H., & He, C. (2024). Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG. Preprints. https://doi.org/10.20944/preprints202405.1656.v1

Chicago/Turabian Style

Liu, X., HongXia Ma and ChengGuang He. 2024 "Prokaryotic Expression and Functional Verification of Antimicrobial Peptide LRGG" Preprints. https://doi.org/10.20944/preprints202405.1656.v1

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 can eliminate 99.99% of E. coli within 100 minutes at the minimal MIC concentration. 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 disruption of Gram-negative cell membrane.

Keywords

Antimicrobial peptides; Prokaryotic expression; Antibacterial mechanism

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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