Antibacterial peptide separated from skin secretions of wood frog and preparation method of antibacterial peptide

Abstract

The invention discloses an antibacterial peptide separated from skin secretions of a wood frog and a preparation method of the antibacterial peptide, and belongs to the technical fields of biomedicine and biology. The antibacterial peptide is characterized in that an amino acid sequence of the antibacterial peptide is SEQ ID No.1 (Sequence Identifier Number 1): Phe-Leu-Pro-Phe-Phe-Ala-Ala-Cys-Ala-Ile-Thr-Lys-Lys-Cys, and the molecular weight of the antibacterial peptide is 1556.8 Dalton. The preparation method of the antibacterial peptide comprises the steps of 1) extracting the skin secretions and 2) separating and purifying, wherein in the step 1), an alive wood frog is adopted, an ear-side gland and a back of the wood frog are stimulated with 10V stabilized voltage discontinuously, and the secretions are collected for later use; in the step 2), ultracentrifugation filtration is performed; a filtrate is collected; liquid chromatography separation is performed; a solution A is trifluoroacetic acid; a solution B is acetonitrile solution containing trifluoroacetic acid; a chromatographic column is balanced with the solution A before being used; after the column is balanced, a filtrate is added to the column; the loading quantity of a sample is 2mL / time; then the solution B is added; a peak with the best antibacterial activity is collected; and the filtrate is further separated by utilizing a reversed phase high performance liquid chromatography. The antibacterial peptide has the beneficial effects that the antibacterial peptide has better resistance to escherichia coli and staphylococcus, and also has an inhibiting effect on clinically separated MRSA (Methicillin-Resistant Staphylococcus Aureus). The antibacterial peptide has the inhibiting effect on MRSA, due to the particularity of an antibacterial mechanism of the antibacterial peptide, no drug resistance can be generated, and the antibacterial peptide can become a novel antibacterial agent resisting to clinical drug-resistant strain or even superbacteria.

Description

technical field [0001] The invention belongs to the fields of biomedicine and biotechnology. Background technique [0002] Antimicrobial peptides, also known as peptide antibiotics or antimicrobial peptides, are widely distributed in viruses, bacteria, insects, and various animals and plants. They are a type of small molecule peptides that can eliminate mutant cells in the body and resist external microorganisms. Antimicrobial peptides not only have broad-spectrum antibacterial activity, but also have high antibacterial efficiency and other advantages, and their biggest advantage is that they are not easy to cause bacterial drug resistance, so they are currently a hot spot in the development and research of anti-pathogenic microbial drugs. [0003] Based on the important role of antimicrobial peptides, many scholars have carried out research on the search and preparation of new antimicrobial peptides. Currently, there are three ways to find and prepare antimicrobial...

AI Technical Summary

Problems solved by technology

Chemically synthesized antimicrobial peptides tend to change their spatial conformation and thus lose their activity

Although the antibacterial peptides obtained by genetic engineering have high yields, there are still problems such as poor stability of expression products and post-translational modifications that may change the activity.

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