Method for producing recombined cationic antibacterial peptide by adopting co-expression anionic ligand

Abstract

The invention discloses a method for producing a recombined cationic antibacterial peptide by adopting a co-expression anionic ligand, which is characterized in that an independent anionic ligand with net negative charges is co-expressed at the same time of expressing the cationic antibacterial peptide, namely, a cationic antibacterial peptide gene and an encoded DNA sequence of the anionic ligand are cloned to a plasmid vector and expressed in a colibacillus host cell in an independent non-fusing way, and the toxicity of the cationic antibacterial peptide to the host cell is restricted by the anionic ligand so as to improve the expression yield; and in the subsequent separation and purification processes, the step of cutting a fusing head, which consumes manpower, physical resources and financial resources, is avoided, therefore, the separation and purification efficiency and the yield ratio are improved. The invention can remarkably shorten the production cycle to one third or a half of the original production cycle and reduce the cost above 50 percent. The invention provides a high-efficiency and low-cost way for the large-scale production of the cationic antibacterial peptide.

Description

Technical field [0001] The invention relates to the field of biological genetic engineering, in particular to a method for co-expressing anionic ligands to produce recombinant cationic antibacterial peptides. Background technique [0002] Cationic antimicrobial peptides (English name cationic antimicrobial peptides, AMPs) are a class of amphiphilic small molecule peptides with a net positive charge. They are widely found in animals, plants and microorganisms. They have a broad bactericidal spectrum and low immunogenicity; Bacterial cell membrane functions to form ion channels on the membrane, causing cell contents to extravasate and die. This unique bactericidal mechanism is not easy to produce resistant strains. However, due to the differences in charge between animal cell membrane components and bacterial cell membrane components, cationic antimicrobial peptides are generally less toxic to animal cells. Against the background that the current drug resistance problem is increas...

AI Technical Summary

Problems solved by technology

If cutting with cyanogen bromide, it is required that the target cationic antimicrobial peptide itself does not contain methionine, and cyanogen bromide itself is also a toxic substance, which may also cause problems such as chemical modification of the antimicrobial peptide; the enzymatic cleavage method requires expensive specific proteases, Excess amino acid residues are left after cleavage (different drugs produced by recombination may therefore have the same redundant amino acid residues, and repeated cross-use will cause immune responses to affect drug efficacy), and non-specific cleavage interference

In a word, both the chemical cleavage method and the enzymatic cleavage method have relatively large limitations and defects in actual production, which have formed a relatively large restriction on the production of cationic antimicrobial peptides by genetic engineering.