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Microbiology and Biotechnology Letters

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Molecular and Cell biology, and Microbial Isolation

Microbiol. Biotechnol. Lett. 2012; 40(2): 92-97

https://doi.org/10.4014/kjmb.1203.03004

Received: March 13, 2012; Accepted: April 3, 2012

Glutathione S-Transferase에 융합한 재조합 Hybrid Peptide Gaegurin-LL37의 대장균에서의 발현

Expression of Recombinant Hybrid Peptide Gaegurin4 and LL37 using Fusion Protein in E. coli

Ishvaanjil Bayarbat 1, Jae-Hag Lee 2 and Soon-Youl Lee 3*

1Department of Chemistry, Mongolian State University of Agriculture, 2Department of Food and Nutrition, Seoil College, 3Department of Biotechnology, Hankyong National University

Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1–16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

Keywords: antimicrobial peptide, LL37, Gaegurin(GGN4),, fusion, heterologous expression, E. coli

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