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Antimicrobial peptide yhx-3 and its composition and application

A YHX-3, antimicrobial peptide technology, applied in the biological field, can solve the problems of low production difficulty, low antibacterial activity, low biological activity, easy to cause hemolysis, etc., achieve chemical synthesis with low difficulty, low hemolytic activity, and save large-scale production costs Effect

Active Publication Date: 2022-03-25
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, most antimicrobial peptides still have problems such as high production cost, low biological activity, and easy hemolysis. Therefore, how to develop antimicrobial peptides with low production difficulty, significant antibacterial activity and no obvious hemolytic activity through rational design Hot Spots in the Research and Development Field of Antimicrobial Peptides

Method used

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  • Antimicrobial peptide yhx-3 and its composition and application
  • Antimicrobial peptide yhx-3 and its composition and application
  • Antimicrobial peptide yhx-3 and its composition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The design of embodiment 1 antimicrobial peptide

[0027] Based on the de novo design of antimicrobial peptides and the understanding of the structure-activity relationship, the sequence parameters of 843 antimicrobial peptide sequences screened from the APD3 database with antibacterial activities against Gram-positive and Gram-negative bacteria were analyzed. Including the analysis of parameters such as sequence length, charged number, hydrophobic amino acid ratio and amino acid composition, the advantageous parameters were selected, and the sequence parameters of the new antimicrobial peptide YHX-3 were determined, as shown in Table 1 below:

[0028] Table 1 Sequence parameters of de novo designed antimicrobial peptides

[0029] sequence length charged number Number of hydrophobic amino acids amino acid composition 13 +5 7 Lys, Ser, Leu

[0030] The length of the amino acid sequence is determined to be 13 in order to improve its safety whi...

Embodiment 2

[0043]Embodiment 2 antibacterial activity detection

[0044] The antimicrobial peptide was synthesized by Qiangyao Biotechnology (Shanghai) Co., Ltd. The C-terminus of the antimicrobial peptide was modified by amidation, and the purity was above 95%.

[0045] Staphylococcus aureus, Escherichia coli and Salmonella were respectively inoculated on LB solid medium, Listeria monocytogenes and Streptococcus mutans were inoculated on BHI solid medium, and cultured in a constant temperature incubator at 37°C After 18 hours, a single colony of each strain was picked and placed in their corresponding liquid medium, and cultured with shaking at a constant temperature of 37°C for 12 hours. Measure the OD of the bacterial solution 600 value, and dilute it to 1×10 6 CFU / mL.

[0046] (1) Inhibition zone test

[0047] Prepare LB and BHI semi-solid medium (mass fraction of agar 0.6%), add 7 μL of bacterial solution to 20mL of each plate, shake and mix well, pour it into the petri dish with...

Embodiment 3

[0053] Example 3 Detection of hemolytic activity

[0054] Take 1mL of healthy rabbit blood and add it to a heparin anticoagulant tube, centrifuge at 1000xg for 10min, get the precipitate, wash with PBS buffer 3 times, and resuspend the red blood cells in 10mL of PBS. Adjust the concentration of antimicrobial peptide YHX-3 to 4-512 μg / mL with PBS buffer, add 50 μL per well into a 96-well plate, and add an equal volume (50 μL) of red blood cell suspension and mix well. Use PBS buffer as a negative control and 0.1% Tritonx-100 as a positive control, take it out after incubating at 37°C for 1 hour, centrifuge at 1000xg for 10 minutes, take out the supernatant and measure the OD value at 570nm with a microplate reader, the test results are shown in the table 4.

[0055] The calculation formula of hemolysis rate is: hemolysis rate=(A T -A O ) / (A C -A O )×100%;

[0056] In the formula, A T is the absorbance value of the experimental group, A C is the absorbance value of the p...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to an antimicrobial peptide YHX‑3 and its composition and application. The amino acid sequence of the antimicrobial peptide YHX‑3 is shown in SEQ ID No.1. The antimicrobial peptide YHX-3 provided by the present invention has broad-spectrum antibacterial activity, and is effective against Gram-positive bacteria and All Gram-negative bacteria have significant antibacterial activity; and their hemolytic activity is low, the sequence length is short, the molecular weight is small, and the chemical synthesis is less difficult. It can kill pathogenic bacteria in organisms more specifically and save scale. Cost of production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antibacterial peptide YHX-3 and its composition and application. Background technique [0002] Since the discovery of penicillin, the first antibiotic, antibiotics have played an important role as a class of anti-infective drugs in the fight against pathogens and saved countless lives. However, in recent years, the abuse of antibiotics has intensified, leading to drug resistance of pathogenic microorganisms, and even the emergence of "super bacteria", which seriously threatens the cause of human health. In 2017, the World Health Organization published the first list of antibiotic-resistant "priority pathogens", listing the 12 species of bacterial species that currently pose the greatest threat to human health, including Acinetobacter baumannii and Pseudomonas aeruginosa. Carbapenems and third-generation cephalosporins produce drug-resistant bacteria, so it is urgent t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K8/64A61K38/10A61P31/04A61Q19/00A01N37/46A01P3/00A01P1/00A23K20/147A23L3/3526
CPCC07K7/08A61K8/64A61P31/04A61Q19/00A01N37/46A23K20/147A23L3/3526A61K38/00A61K2800/524A23V2002/00A23V2200/10A23V2250/55Y02A50/30
Inventor 易华西李剑勋公丕民刘同杰张兰威韩雪步雨珊
Owner OCEAN UNIV OF CHINA
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