Preparation method and application of novel antimicrobial peptide Misgurin mutant

Abstract

The invention belongs to the technical field of biology and relates to a preparation method and application of a novel antimicrobial peptide Misgurin mutant. The technical scheme is as follows: introducing glutamine at the C tail end of the amino acid sequence of natural Misgurin so as to design a novel Misgurin mutant; selecting Pichia yeast biased codons so as to artificially synthesize a complete gene of the novel Misgurin mutant; constructing a Pichia yeast secretion type expression vector pPICZaA-M; transforming a host cell SMD1168 by a vector electroporation method; selecting to obtain high-efficiency expression strains; and under proper culture conditions, ensuring 100 microgram of recombinant novel Misgurin mutant is contained in per milliliter of expression supernatant. The obtained product has a good nematocidal activity on staphylococcus aureus, pig salmonella and pig pathogenic Escherichia coli K88, K99 and 987P, and has an obvious effect on prevention and treatment of clinical bacterial diseases.

Description

technical field [0001] The invention belongs to the high-tech research field of biotechnology, and relates to a novel antimicrobial peptide Misgurin mutant, more specifically to a preparation method of a recombinant novel antimicrobial peptide Misgurin mutant and its application in controlling and preventing bacterial diseases. Background technique [0002] Antibacterial peptides (ABP) are cationic small peptides produced by organisms with strong antibacterial effects, and are important components of biological innate immunity. The mechanism of action of antimicrobial peptides is different from that of traditional antibiotics, and they are the most promising substitutes for antibiotics. Compared with traditional antibiotics, antimicrobial peptides have a unique antibacterial mechanism, which is not easy to cause drug-resistant mutations in bacteria. Antimicrobial peptides can also specifically inhibit fungi, viruses, parasites and tumors, and their biological activities are...

AI Technical Summary

Problems solved by technology

In 2002, Gong Jiewan et al. concatenated multiple copies of the Misgurin gene in the same direction, cloned it into the expression plasmid pPIC9K, and electrotransformed it into Pichia pastoris strain GS115, and screened out recombinants that integrated multiple copies of the foreign gene. The antibacterial activity of the recombinant protein was not mentioned in the paper (Jiewan Gong. The construction of gene expression vectors for mae and misgurin and their expression in Pichia pastoris. Jinan University, Ph.D. dissertation, 2002.48-57.)

At present, there is no report on the successful use of genetic engineering technology to produce active antimicrobial peptide Misgurin, and there is no report on the commercialization of new antimicrobial peptide Misgurin mutants

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