Efficient hybrid antibacterial peptide LI and preparation method and application thereof

An antimicrobial peptide and high-efficiency technology, which is applied in the field of high-efficiency hybrid antimicrobial peptide LI and its preparation and application, can solve the problems of small molecular weight, low activity and complex structure of antimicrobial peptides

Inactive Publication Date: 2017-02-22
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2. Production cost of antimicrobial peptides
Blindly using genetic engineering to express antimicrobial peptides without comprehensively exploring the structure and function of antimicrobial peptides and in vitro tests, resulting in the expression of antimicrobial peptides with low activity or cytotoxicity such as hemolysis
[0003] Antimicrobial peptides have the smallest molecular weight, various types and complex structures

Method used

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  • Efficient hybrid antibacterial peptide LI and preparation method and application thereof
  • Efficient hybrid antibacterial peptide LI and preparation method and application thereof
  • Efficient hybrid antibacterial peptide LI and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The amino acid sequence of human antimicrobial peptide LL37 is:

[0017]

[0018] The amino acid sequence of bovine antimicrobial peptide In13 is:

[0019] Ile Leu Pro Trp Lys Trp Pro Trp Trp Pro Trp Arg Arg-NH2

[0020] 1 5 10 13

[0021] According to the site-specific amino acid fragment interception method, the amino acid residues 14-21 of human antimicrobial peptide LL37 (GlyLysGluPheLysArgIleVal) and the fragments of amino acid residues 5-13 of bovine antimicrobial peptide In13 (LysTrpProTrpTrpProTrpArgArg) were intercepted.

[0022] The above two fragments were sequentially connected to obtain LI (GlyLysGluPheLysArgIleValLysTrpProTrpTrpProTrpArgArg-NH 2 ) antimicrobial peptides.

Embodiment 2

[0024] The above two antimicrobial peptides were synthesized using a peptide synthesizer. The method was solid-phase chemical synthesis, and the specific steps were:

[0025] 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;

[0026] 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

[0032] 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 efficient hybrid antibacterial peptide LI and a preparation method and application thereof. A sequence of the efficient hybrid antibacterial peptide LI is shown as SEQ ID No.1 of a sequence table. The preparation method includes following steps: according to a fixed-point amino acid fragment intercepting method, intercepting an amino acid residue fragment from the 14th locus to the 21st locus of human-derived antibacterial peptide LL37 and an amino acid residue fragment from the 5th locus to the 13th locus of bovine-derived antibacterial Indolicidin, and sequentially connecting the two fragments to obtain the antibacterial peptide LI; 2), using a polypeptide synthesizer for solid-phase synthesis to synthesize peptide resin; obtaining polypeptide after TFA cutting, and adopting reverse-phase high-performance liquid chromatography for purifying. The antibacterial peptide LI obtained by the method has high antibacterial activity, weak hemolytic activity, high treatment index and huge development potential.

Description

technical field [0001] The invention relates to a high-efficiency hybrid antibacterial peptide LI and its preparation method and application. Background technique [0002] Antimicrobial peptides are an animal's first line of defense against exogenous pathogens. It not only has the function of killing pathogenic bacteria, but also has biological functions such as anti-fungal, virus, parasite and anti-cancer cells. Antimicrobial peptides have a unique bactericidal mechanism, making them the most promising alternatives to antibiotics. Antimicrobial peptide molecules have positively charged amino acids, such as arginine, lysine, and histidine, and hydrophobic amino acids with hydrophobic residues, such as leucine, isoleucine, valine, tryptophan, Phenylalanine, alanine, etc. Antibacterial peptides with electrostatic positive charges and negatively charged bacterial cell membranes attract each other electrostatically, antibacterial peptides are close to bacterial cells, and hyd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K1/20C07K1/04A61K38/10A61P31/04
CPCA61K38/00C07K14/4723C07K2319/00
Inventor 董娜刘春玉李欣然
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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