Heterologous expression of the novel dimeric antimicrobial peptide LIG in Pichia pastoris.

The antimicrobial peptide (AMP) LI is a fusion product of antimicrobial peptide LL37 produced by human neutrophils and Indolicidin secreted by bovine neutrophils. LI retained the antimicrobial activity of the parental peptides and showed high cell selectivity. In this study, the flexible linker Gly-Ser-Gly (G-S-G) was used to ligate LI into dimeric LIG, and constructed the Pichia pastoris (P. pastoris) expression vector pPIC9K-6×His-3×FLAG-LIG. The total protein expression of P. pastoris GS115 reached the highest level (189.6mg/L) after 96h induction with 3% methanol at the initial pH value of 7.0. Finally, 5.9mg/L of recombinant LIG (rLIG) was obtained after enterokinase digestion and purification. The rLIG had high antimicrobial activity and low hemolytic activity. Compared with monomer LI, GSG linked dimeric LIG, which had no significant change in antimicrobial activity and had good salt ions stability. In this study, the dimeric antimicrobial peptide LIG was successfully expressed, which provided a new idea for the expression of AMPs in the P. pastoris expression system, and had important significance for the application of AMPs.

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Copyright © 2024. Published by Elsevier B.V.

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Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. Dong reports financial support was provided by Foundation for Innovative Research Groups of the National Natural Science Foundation of China. Dong reports financial support was provided by the National Key R & D Program of China. Dong reports financial support was provided by the China Agriculture Research System. Dong reports financial support was provided by the Science Fund for Distinguished Young Scholars of Heilongjiang Province.