Preparation method and activity detection for antibacterial peptides

A technology for activity detection and antibacterial activity, applied in the field of molecular biology, can solve the problems of low antimicrobial peptide activity and high production cost, and achieve the effect of low hemolysis and cytotoxicity and high antibacterial activity

Inactive Publication Date: 2010-10-06
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
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AI Technical Summary

Problems solved by technology

However, there are still some problems and bottlenecks in the research of antimicrobial peptides, mainly as follows: first, although many antimicrobial peptides have spectrum antibacterial properties, they also cause hemolysis to normal cells, such as red blood cells; second, many antimicrobial peptides Composed of more than 20 amino acids, the production cost of solid-phase synthesis is too high; third, blindly using genetic engineering methods to express antimicrobial peptides without comprehensive exploration of the structure and function of antimicrobial peptides and in vitro tests, resulting in poor expression Most of the antimicrobial peptides are not highly active, or have hemolytic activity and cytotoxicity

Method used

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  • Preparation method and activity detection for antibacterial peptides
  • Preparation method and activity detection for antibacterial peptides
  • Preparation method and activity detection for antibacterial peptides

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Embodiment 1

[0008] Embodiment 1: The preparation method and activity detection of an antibacterial peptide of the present invention, the preparation method is as follows: adopt the spiral wheel model of protein to design 4 kinds of antibacterial peptides with antibacterial activity, wherein in the polar plane, used 5 arginines Or lysine provides a positive charge for the antimicrobial peptide, so that the antimicrobial peptide can be attracted to the negative charge on the surface of the bacterial cell membrane, while on the non-polar surface, 8 leucine or valine are used to provide hydrophobic groups for the antimicrobial peptide , so that the antimicrobial peptide can interact with the lipid bilayer of the bacterial cell membrane, and then form holes on the bacterial cell membrane, and finally make the bacteria be inhibited or die. In order to reduce the hemolytic activity of the antimicrobial peptide, it is also increased by introducing 3 glycines The flexibility of the antimicrobial pe...

Embodiment 2

[0009] Embodiment 2: the preparation of antimicrobial peptide

[0010] According to the spiral wheel model, 5 arginines or lysines are used as polar ends to provide positive charges, so that they interact with the negative charges on the surface of the bacterial membrane through physical attraction; 8 leucines or valines are used as The residues at the non-polar end make it interact with the non-polar group of the lipid bilayer of the bacterial cell membrane, thereby destroying the bacterial cell membrane, causing it to collapse and die. At the same time, in order to reduce the toxicity of antimicrobial peptides to normal cells (such as red blood cells or mammalian cells) and increase the flexibility of this model, three additional glycines were added to the structure.

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Abstract

The invention provides a preparation method and activity detection for antibacterial peptides. In the preparation method, series of antibacterial peptides which contain a few amino acid residues are prepared by using a helical wheel model of a protein; and antibacterial activity, hemolytic activity, cytotoxicity and the like of the series of the antibacterial peptides are detected. The four obtained antibacterial peptides have obvious bacteriostatic activity; and the two antibacterial peptides which contain a plurality of valines have no hemolytic activity and less cytotoxicity, wherein in particular the antibacterial peptides which contain three glycines, five arginines and eight valines have the greatest potential. The preparation method and the activity detection have important theoretical and practical significance for further explaining the action mechanism of the antibacterial peptides, designing the antibacterial peptides reasonably and the like.

Description

(1) Technical field [0001] The invention relates to molecular biology, in particular to a preparation method and activity detection of an antibacterial peptide. (2) Background technology [0002] Due to the problem of drug residue and bacterial drug resistance of antibiotics, it is urgent to find a green, pollution-free and non-bacterial drug-resistant antibiotic substitute. Antimicrobial peptide is an active polypeptide with antibacterial effect that widely exists in organisms. It is the product of immune response of biological non-specific defense system. It has broad-spectrum antibacterial activity. It can not only inhibit a variety of bacteria or fungi, but even inhibit the growth of tumor cells. Antimicrobial peptides produce antibacterial or bactericidal effects mainly through physical penetration on bacterial cell membranes, and it is difficult for microorganisms such as bacteria to change the cell membrane structure of their own phospholipid bilayers, thus making ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/04C12Q1/18C12Q1/02G01N33/68G01N21/31
Inventor 单安山马清泉郑燕斌
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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