Octanoic acid acylation modified antibacterial peptide and application thereof

An antibacterial peptide and amino acid technology, applied in the fields of genetic engineering and biological preparations, can solve the problems of poor activity, insufficient antibacterial activity of fungi, and high toxicity

Active Publication Date: 2021-05-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a large number of antimicrobial peptides are currently isolated from animals, natural antimicrobial peptides have the disadvantages of poor activity and high toxicity, especially the lack of antibacterial activity against fungi

Method used

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  • Octanoic acid acylation modified antibacterial peptide and application thereof
  • Octanoic acid acylation modified antibacterial peptide and application thereof
  • Octanoic acid acylation modified antibacterial peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] In this embodiment, the antimicrobial peptide of the present invention (see SEQ ID NO.1) is synthesized by solid-phase chemical synthesis, and the peptide structure is as follows: figure 1 Shown:

[0029] 1. The preparation of antimicrobial peptides is carried out one by one from the C-terminal to the N-terminal, 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;

[0030] 2. Add a cleavage reagent to the peptide resin obtained above, react for 2 hours at 20°C in the dark, filter; wash the precipitate wi...

Embodiment 2

[0034] Determination of the antifungal activity of the peptide: the minimum inhibitory concentration of the peptide (C8LR) prepared in Example 1 against fungi was determined by microdilution method. Add 0.2% fetal bovine serum albumin containing 0.01% acetic acid as a diluent into a 96-well plate, and use the serial dilution method to configure the antibacterial peptide solution obtained in Example 1 of the present invention in sequence, so that the solution in each well is The volume is 50 μL. Then add 50 μL of the bacteria solution to be tested (~10 3 CFU / mL) in each well, the medium is RPMI 1640 (pH=7.0) containing MOPS. Positive controls (containing bacterial fluid but not antimicrobial peptides) and negative controls (neither bacterial fluid nor peptides) were set up. Incubate at a constant temperature of 28°C for 48 hours, then measure the light absorption value at 492nm with a microplate reader, and use the value greater than 0.1 as the criterion for the growth of the...

Embodiment 3

[0039] Determination of the antibacterial activity of the peptide: the minimum inhibitory concentration of the peptide (C8LR) prepared in Example 1 against bacteria was determined by microdilution method. Add 0.2% fetal bovine serum albumin containing 0.01% acetic acid as a diluent into a 96-well plate, and sequentially prepare a series of gradient antimicrobial peptide solutions using the doubling dilution method, so that the volume of the solution in each well is 50 μL. Then add 50 μL of the bacteria solution to be tested (~10 5 CFU / mL) in each well, the medium is MHB (pH=7.0). Positive controls (containing bacterial fluid but not antimicrobial peptides) and negative controls (neither bacterial fluid nor peptides) were set up. Incubate at a constant temperature of 37°C for 18 hours, then measure the light absorption value at 492nm with a microplate reader, and use the value greater than 0.1 as the judgment standard for the growth of the strain to determine the minimum inhib...

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PUM

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Abstract

The invention relates to the technical field of genetic engineering and biological agents, and particularly discloses octanoic acid acylation modified antibacterial peptide and application thereof. The amino acid sequence of the antibacterial peptide is shown as SEQ ID NO. 1. The octanoic acid acylation modified antibacterial peptide simulates the arrangement rule of alpha-spiral secondary structure amino acid in natural protein, leucine and arginine provide hydrophobicity and cationicity respectively, a heptapeptide repetitive sequence 'abcdefg' is used as a template, the sequence is repeated twice, then the tail end of the sequence N is subjected to octanoic acid acylation modification, and the tail end of the sequence C is amidated. The peptide has broad-spectrum antibacterial activity on fungi and bacteria and has wide clinical application potential.

Description

technical field [0001] The invention relates to the technical fields of genetic engineering and biological preparations, in particular to an octanoic acid acylated antimicrobial peptide and its application. Background technique [0002] The abuse of antibiotics has led to the emergence of drug-resistant fungi and bacteria, especially superbugs, which have developed resistance to multiple antibiotics and seriously threatened human health. Although traditional antifungal drugs such as ketoconazole and fluconazole have certain therapeutic effects, they are more likely to induce drug resistance in fungi and cause certain side effects on the human body. Therefore, there is an urgent need to develop a novel antifungal drug with low toxicity and high efficiency. [0003] Antimicrobial peptides exist in almost all animals and constitute the first line of defense of the animal body. Usually, antimicrobial peptides exert antibacterial activity by disturbing the membrane stability of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/10A61P31/04A23L33/18A23L3/3526
CPCC07K7/08A61P31/10A61P31/04A23L33/18A23L3/3526A61K38/00A23V2002/00A23V2200/10A23V2200/30A23V2250/55Y02A50/30
Inventor 马曦谭鹏唐琪张钰成
Owner CHINA AGRI UNIV
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