Antibacterial peptide LL-37-Melittin, application of antibacterial peptide, and preparation method of antibacterial peptide in bacillus subtilis

Abstract

The invention discloses antibacterial peptide LL-37-Melittin, an application of the antibacterial peptide, and a preparation method of the antibacterial peptide in bacillus subtilis. The method comprises the steps of expressing the antibacterial peptide LL-37-Melittin in the bacillus subtilis by utilizing a fusion technology, manually designing a fusion gene SUMO-LL-37-Melittin, adding EcoRI and BamHI restriction enzyme cutting sites at the two ends, connecting the synthetized fusion gene and an expression vector pGJ148, successfully constructing a recombinant expression vector, transforming the recombinant expression vector into the bacillus subtilis WB800N to obtain an expression engineering bacterium, performing maltose induction expression in a culture medium, purifying an expression product by Ni-NAT affinity chromatography, detecting the correctness of the expression product by utilizing SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and performing SUMO protease restriction enzyme cutting to obtain the antibacterial peptide LL-37-Melittin. The method expresses the antibacterial peptide in the bacillus subtilis; separation and purification are simple; and the method is easy to operate. The expression product has antibacterial activity for multiple bacteria, is good in thermal stability, is suitable for large-scale industrial production, and can be applied in the fields of medicines, food, animal husbandry and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antibacterial peptide LL-37-Melittin, its application and a preparation method in Bacillus subtilis. Background technique [0002] Antimicrobial peptides are part of the innate immune mechanism of animals, plants and humans. They have a broad-spectrum antibacterial effect and have a killing effect on some pathogenic bacteria that are resistant to antibiotics. So far, thousands of natural antibacterial peptides have been discovered, but there are relatively few species with good medical selection index. Ideal new artificial antibacterial peptides were obtained from peptide resources. [0003] The recombinant expression of antimicrobial peptides faces two huge problems. First, the antimicrobial properties of antimicrobial peptides make them potentially lethal to host bacteria. Second, the small molecular weight and high cationicity of antimicrobial peptides make them ...

AI Technical Summary

Problems solved by technology

First, the antimicrobial properties of antimicrobial peptides make them potentially lethal to host bacteria

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