Antibacterial peptide GW13 and its preparation method and use

A technology of antimicrobial peptides and peptide resins, applied in the field of antimicrobial peptides

Active Publication Date: 2012-12-19
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports and studies on the relationship between the structure and function of antimicrobial peptides using the small peptide-li...

Method used

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  • Antibacterial peptide GW13 and its preparation method and use
  • Antibacterial peptide GW13 and its preparation method and use
  • Antibacterial peptide GW13 and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] The amino acid sequence of linear chicken β-defensin 4 is:

[0014] Arg Tyr His Met Gln Cys Gly Tyr Arg Gly Thr Phe Cys Thr Pro

[0015] 1 5 10 15

[0016] Gly Lys Cys Pro Tyr Gly Asn Ala Tyr Leu Gly Leu Cys Arg Pro;

[0017] 20 25 30

[0018] Lys Tyr Ser Cys Cys Arg Trp Leu-NH2

[0019] 35 38

[0020] Targeted interception of 13 amino acid fragments at the carboxy-terminal of linear chicken β-defensin 4, and removal of the disulfide bond to obtain GW10 (GlyLeuArgProLysTyrSerArgTrpLeu-NH 2 ) fragment.

[0021] The typical fragment ArgTrpLeu was repeatedly connected to obtain GW13(GlyLeuArgProLysTyrSer(ArgTrpLeu) 2 -NH 2 ) antimicrobial peptides. GW10 and GW13 are small peptides with 10 and 13 amino acids.

Embodiment 2

[0023] The above two antimicrobial peptides were synthesized using a peptide synthesizer. The method was solid-phase chemical synthesis, and the specific steps were:

[0024] 1) The preparation of antimicrobial peptides is carried out one by one from the C-terminus to the N-terminus, and is completed by a peptide synthesizer. First, Fmoc-X (X is the first amino acid at the C-terminal of each antimicrobial peptide) is inserted into Wang resin, and then the Fmoc group is removed to obtain X-Wang resin; then Fmoc-Y-Trt-OH (9 -Fmoxy-trimethyl-Y, Y is the second amino acid at the C-terminus of each antimicrobial peptide); according to this procedure, it is synthesized from the C-terminus to the N-terminus until the synthesis is completed, and the side of the Fmoc group is removed chain protection resin;

[0025] Add cutting reagent to the peptide resin obtained above, react at 20°C for 2 hours in the dark, filter; precipitate TFA (trifluoroacetic acid) for washing, mix the washing...

Embodiment 3

[0031] The designed and synthesized antimicrobial peptides were compared and detected by in vitro antibacterial and hemolytic activity tests;

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Abstract

The invention relates to an antibacterial peptide GW13 and its preparation method and use. The antibacterial peptide GW13 has an amino acid sequence of GlyLeuArgProLysTyrSer(ArgTrpLeu)2-NH2. The preparation method comprises the following steps of 1, cutting out 13 amino acid fragments from a linear chicken beta-defensin 4 carboxyl end by a fixed-point amino acid fragment cutting method, removing disulfide bonds to obtain a GW10 (GlyLeuArgProLysTyrSerArgTrpLeu-NH2) fragment, and repeating connecting characteristic three-residue complexes ArgTrpLeu to obtain the antibacterial peptide GW13, and 2, synthesizing a peptide resin in a polypeptide synthesizer by a solid-phase synthesis method, carrying out TFA cutting to obtain a polypeptide, and carrying out reversed-phase high-performance liquid chromatography purification. The antibacterial peptide GW13 obtained by the preparation method has strong bacteriostatic activity, low hemolytic activity, the highest therapeutic index and a large development potential.

Description

technical field [0001] The invention relates to an antimicrobial peptide, in particular to an antimicrobial peptide GW13 and its preparation method and application. Background technique [0002] Antimicrobial peptides are active polypeptides with antibacterial effects that widely exist in organisms. They are immune response products of biological nonspecific defense systems and have broad-spectrum antibacterial, antiviral, antifungal, antiparasitic and antitumor biological activities. The bacteriostatic mechanism of antimicrobial peptides is different from that of antibiotics. It acts on the bacterial cell membrane through physical penetration, destroying the phospholipid bilayer on the membrane, so that the substances in the membrane flow out, resulting in the death of bacteria. Therefore, antimicrobial peptides do not produce drug resistance. Antimicrobial peptide itself is a kind of polypeptide substance, and there is no residual problem. Therefore, antimicrobial peptide ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/06C07K1/04A61K38/10A61P31/04
Inventor 单安山董娜马清泉胡婉宁
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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