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A Novel Antimicrobial Peptide and Its Application

A technology of antibacterial peptides and antibacterial ingredients, applied in the application, antibacterial, antifungal and other directions, can solve the problems of high toxicity, poor stability, low antibacterial activity, etc., and achieve strong antibacterial properties and good stability.

Active Publication Date: 2022-08-09
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, quite a few natural antimicrobial peptides have low antibacterial activity, poor stability, high toxicity, and cause hemolysis of eukaryotic cells; in addition, some natural antimicrobial peptides have poor inhibitory effect on drug-resistant bacteria and cannot meet the requirements of practical applications.
There is no report about the use of this short peptide other than drug carrier

Method used

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  • A Novel Antimicrobial Peptide and Its Application
  • A Novel Antimicrobial Peptide and Its Application
  • A Novel Antimicrobial Peptide and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Antibacterial peptides inhibit the growth of different microorganisms

[0046] Material of the present invention: Ac-Ala-Ala-Ala-Ala-Ala-Ala-Pro-Lys-Lys-Pro-Ala-Ala-Ala-Ala-Ala-Ala-NH 2 (SEQ ID NO.1), abbreviated as APK, was entrusted to Shanghai Botai Biotechnology Co., Ltd. for chemical synthesis. Among them, N-terminal acetylation (adding acetyl Ac) and C-terminal amidation (adding amino NH) 2 ) is also known as the blocking of the peptide end, which can shield the dissociable groups at the two free ends of the polypeptide and increase its end hydrophobicity.

[0047] A single clone of Escherichia coli (E.coli, LB medium), Staphylococcus aureus (S.aureus, LB medium), and Candida albicans (C.albicans, YPD medium) was cultured at 37°C and 180rpm overnight. Then 7100rpm, 2min centrifugation to collect bacterial cells, dilute E.coli and S.aureus to OD600 of 0.4 and C.albicans to 0.8, and then inoculate the bacterial solution on a 96-well plate at 5% per well...

Embodiment 2

[0050] Example 2: Antibacterial peptides inhibit the growth of different microorganisms

[0051] Material of the present invention: Ac-Ala-Ala-Ala-Ala-Ala-Ala-Pro-Arg-Arg-Pro-Ala-Ala-Ala-Ala-Ala-Ala-NH 2(SEQ ID NO.2), Ac represents acetyl group, abbreviated as APR; and entrusted Shanghai Botai Biotechnology Co., Ltd. to carry out chemical synthesis.

[0052] A single clone of Escherichia coli (E.coli, LB medium), Staphylococcus aureus (S.aureus, LB medium), and Candida albicans (C.albicans, YPD medium) was picked and cultured at 37°C and 180rpm overnight. Then 7100rpm, 2min centrifugation to collect the bacteria, dilute E.coli and S.aureus to OD600 of 0.4 and C.albicans to 0.8, and then inoculate the bacteria solution on a 96-well plate at 5% per well containing 0, 50 μM, 200 μM and 400 μM APK were cultured in 100 μL medium at 37°C and 70 rpm, and the OD600 was measured with a microplate reader at different time points, and the growth curve was drawn.

[0053] from figure ...

Embodiment 3

[0055] Example 3: Antibacterial peptides self-assemble to form nanofibers

[0056] Materials: APK and APR short peptides were entrusted to Shanghai Botai Biotechnology Co., Ltd. for chemical synthesis.

[0057] The APK and APR short peptides were dissolved in deionized water to prepare a 1 mM solution. 10 μL of the short peptide solution was taken and dropped on a 400-mesh copper mesh for 5 min, and then the liquid was removed with filter paper. Then stain with 10 μL of 2% phosphotungstic acid for 2 min, absorb the liquid with filter paper and air dry naturally. The nanostructures formed by the self-assembly of APK and APR were observed by TEM.

[0058] from image 3 It can be seen that both APK and APR self-assemble in solution to form nanofibers.

[0059] The results of Example 3 show that the nanofibers formed by the self-assembly of APK and APR may be the main structure that encapsulates microorganisms and inhibits their growth and proliferation.

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Abstract

The invention relates to a novel antibacterial peptide and its application, and belongs to the field of antibacterial peptides. The involved antimicrobial peptide amino acid sequence is (Y)n‑Pro‑X‑X‑Pro‑(Y)n. Wherein X is lysine or arginine, Y is hydrophobic amino acid, n is the number of hydrophobic amino acid, and the value range is 4-8. The antibacterial peptide of the present invention can significantly inhibit various bacteria and fungi such as Escherichia coli, Staphylococcus aureus, Candida albicans, etc., and has very low hemolytic activity. The antibacterial peptide of the present invention has excellent effect after experimental verification, and has great development prospect.

Description

technical field [0001] The present invention belongs to the field of antimicrobial peptides. Background technique [0002] As an important clinical drug, traditional antibiotics have achieved remarkable results since their application. But in recent years, the abuse of antibiotics and the emergence of drug-resistant bacteria, especially the emergence of multi-drug-resistant bacteria (super bacteria), have become one of the main problems faced by the medical field in the world today. It has also prompted the search for alternatives to antibiotics. Polypeptide antibacterial drugs are considered to have broad application prospects in the pharmaceutical industry due to their high antibacterial activity and resistance to drug resistance. They are also the hope for people to solve drug-resistant bacteria and promote human health. [0003] Antimicrobial peptides are a class of small-molecule polypeptides that are widely distributed in animals and plants and are part of the natura...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/08A61P31/04A61P31/10A61K8/64A61Q17/00A01N37/46A01P1/00A01P3/00C11D3/48
CPCA61K38/08A61P31/04A61P31/10A61K8/64A61Q17/005A01N37/46C11D3/48A61K2800/10Y02A50/30
Inventor 邱峰彭飞张文胜
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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