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http://dx.doi.org/10.4014/kjmb.1203.03004

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

Bayarbat, Ishvaanjil (Department of Chemistry, School of Natural Science, Mongolian State University of Agriculture)
Lee, Jae-Hag (Department of Food and Nutrition, Seoil College)
Lee, Soon-Youl (Department of Biotechnology, Genetic Engineering Research Institute, Hankyong National University)
Publication Information
Microbiology and Biotechnology Letters / v.40, no.2, 2012 , pp. 92-97 More about this Journal
Abstract
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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