High-stability antibacterial peptide and application thereof

An antibacterial peptide and acid technology, applied in the fields of genetic engineering and biological preparations, can solve the problems of increasing production cost, complex application environment, hindering the application of peptides, etc., and achieves the effects of low cost, low hemolytic activity and enhanced activity

Active Publication Date: 2021-04-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these modified peptides can only be obtained by chemical synthesis, which greatly increases the production cost of the peptide, and the high price hinders the further application of the peptide
[0004] In past studies, researchers often only considered the stability in one medium, and the application environment of antimicrobial peptides is complex when they enter the body, and they are usually subjected to the combined action of proteases and various salts to inactivate antimicrobial peptides

Method used

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  • High-stability antibacterial peptide and application thereof
  • High-stability antibacterial peptide and application thereof
  • High-stability antibacterial peptide and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 solid-phase chemical synthesis method synthesizes antimicrobial peptide of the present invention

[0030] In this embodiment, the antimicrobial peptide of the present invention (see SEQ ID NO.1) is synthesized by solid-phase chemical synthesis method, and the peptide structure schematic diagram is as follows figure 1 As shown, the schematic diagram of the spiral wheel is shown in figure 2 shown.

[0031] 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 co...

Embodiment 2

[0035] The mensuration of the antibacterial activity of embodiment 2 peptide

[0036] The minimum inhibitory concentration of the peptide (RP29) prepared in Example 1 to bacteria was determined by the 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 5CFU / mL) in each well, the medium is MHB (pH=7.4 or 6.5). 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 de...

Embodiment 3

[0043] The mensuration of the hemolytic activity of embodiment 3 peptide

[0044] Take 1 mL of fresh porcine red blood cells and dilute them 10 times with PBS for later use. PBS was used as a diluent to add to a 96-well plate, and a series of gradient antimicrobial peptide (peptide RP29 prepared in Example 1) solutions were sequentially prepared using the doubling dilution method, so that the volume of the solution in each well was 50 μL. Add 50 μL of erythrocyte suspension to the wells containing the peptide, the erythrocyte suspension treated with 0.1% Triton was used as a positive control, and the untreated erythrocyte suspension was used as a negative control. The 96-well plate was then incubated in a 37°C incubator for 1 h. Centrifuge at 1000g for 5 minutes, draw 50 μL of supernatant from each well of the 96-well plate, transfer it to a new 96-well plate, and then use a microplate reader to measure the light absorbance at 570 nm. The test set two parallel ,repeat three ...

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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 a high-stability antibacterial peptide and application thereof. The amino acid sequence of the antibacterial peptide disclosed by the invention is shown as SEQ ID NO. 1. The antibacterial peptide disclosed by the invention is simple in the synthesis method, the amino acids in the sequence are all natural amino acids, the cost is low, and the peptide sequence has high stability in a simulated in-vivo salt ion environment and a simulated trypsin chymotrypsin environment. The antibacterial peptide shows broad-spectrum antibacterial activity under a neutral pH value, and shows an inhibition effect on strains such as escherichia coli, staphylococcus epidermidis, salmonella and staphylococcus aureus. And under an acidic pH condition, the activity is further enhanced. Meanwhile, the antibacterial peptide has relatively low hemolytic activity and good biocompatibility.

Description

technical field [0001] The invention relates to the technical field of genetic engineering and biological preparations, in particular to a highly stable antibacterial peptide and its application. Background technique [0002] Since the discovery of penicillin, antibiotics have saved countless lives and extended the average human lifespan by 24 years. Antibiotics in animal husbandry can improve animal performance and have a growth-promoting effect. However, the frequent abuse of antibiotics will affect the immunity of animals and induce antibiotic-sensitive strains to develop drug resistance. Antibiotic residues in animals will be transmitted to humans through various media, leading to the emergence of drug-resistant bacteria in humans and seriously endangering human health. Therefore, the development of alternatives to antibiotics has become a research hotspot. [0003] Antimicrobial peptides have been studied for many years as drugs to address drug-resistant bacterial in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47A01N47/16A01P1/00A01P3/00A61K38/16A61P31/04A23L33/18
CPCY02A50/30
Inventor 马曦谭鹏张钰成唐琪
Owner CHINA AGRI UNIV
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