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Antibacterial peptide YHX-4 and application thereof

A YHX-4, antibacterial peptide technology, applied in applications, antibacterial drugs, peptides, etc., to achieve the effects of small molecular weight, short synthetic sequence, and low difficulty in chemical synthesis

Active Publication Date: 2022-05-20
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For the above-mentioned problems that existing antimicrobial peptides cannot meet the needs of clinical treatment, the present invention provides an antimicrobial peptide YHX-4 and its application. The antimicrobial peptide of the present invention is effective against Gram-positive bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans), Gram-negative bacteria (Escherichia coli, Salmonella) have good antibacterial activity, and the structure is simple, the synthesis difficulty is small, and the safety is good

Method used

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  • Antibacterial peptide YHX-4 and application thereof
  • Antibacterial peptide YHX-4 and application thereof
  • Antibacterial peptide YHX-4 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The design of embodiment 1 antimicrobial peptide

[0031] 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 ADP3 database with antibacterial effects on Gram-positive and Gram-negative bacteria were analyzed ( Including sequence length, charged number, hydrophobic amino acid ratio and amino acid composition), and then according to the principle of superior parameter selection and combined with rational design ideas, the sequence parameters of the new antimicrobial peptide were determined, as follows:

[0032] Table 1 Sequence parameters of de novo designed antimicrobial peptides

[0033]

[0034] In order to reduce the synthesis cost and reduce the cytotoxicity, the sequence length was selected as 13, which is the second most frequently used sequence. The positively charged amino acid Lys and the polar uncharged amino acid...

Embodiment 2

[0044] Embodiment 2 antibacterial activity

[0045] Antimicrobial peptides were synthesized by Qiangyao Biotechnology (Shanghai) Co., Ltd.

[0046] 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.

[0047] ①Inhibition zone test

[0048] 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 Oxford cup placed, and pull out the Oxford cup after the medium is cooled and solidified to complete the infusion....

Embodiment 3

[0053] Example 3 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-4 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. Take PBS buffer as a negative control, and 0.1% Tritonx-100 as a negative 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, and 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 positive...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an antibacterial peptide YHX-4 and application thereof. Aiming at the problem that the existing antibacterial peptide cannot meet the clinical treatment requirement, the invention provides the antibacterial peptide YHX-4 and the application thereof. The amino acid sequence of the antibacterial peptide YHX-4 is as follows: Trp-Lys-Leu-Leu-Ser-Lys-Leu-Leu-Gly-Lys-Leu-Leu-Lys-NH2, and the amino acid sequence of the antibacterial peptide YHX-4 is as follows: Trp-Lys-Leu-Leu-Lys-NH2. The antibacterial peptide disclosed by the invention has relatively good antibacterial activity on listeria monocytogenes, staphylococcus aureus, streptococcus mutans, escherichia coli and salmonella. The antibacterial peptide is simple in structure, low in synthesis difficulty and good in safety.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antibacterial peptide YHX-4 and its application. Background technique [0002] The widespread use of traditional antibiotics, while providing a powerful weapon for humans to fight against pathogens, has also brought about serious drug resistance problems. According to the data released by the China Bacterial Resistance Trial, bacterial drug resistance is increasing year by year, and some bacteria exhibit multi-drug resistance and cross-resistance, posing a serious threat to the treatment of clinical infections. Therefore, it is urgent to solve the problem of drug resistance of pathogenic bacteria. The development of new antibacterial drugs is an important way to solve this problem. Among them, antimicrobial peptides are not easy to produce drug resistance due to their unique membrane-penetrating mechanism and rapid bactericidal efficiency, and have become the most po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/04A61K8/64A61Q19/10A61Q5/02A61Q11/00A23K20/147A23L3/3526C11D3/37C11D3/48
CPCC07K7/08A61P31/04A61K8/64A61Q19/10A61Q5/02A61Q11/00A23K20/147A23L3/3526C11D3/3719C11D3/48A61K38/00A61K2800/524A23V2002/00A23V2200/10A23V2250/55Y02A50/30
Inventor 易华西李剑勋刘银雪步雨珊刘伊索公丕民刘同杰张兰威
Owner OCEAN UNIV OF CHINA
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