A kind of low molecular weight antimicrobial peptide yhx-2 and its composition and application

A YHX-2, low-molecular-weight technology, which is applied in food ingredients as antimicrobial preservation, application, antibacterial drugs, etc., can solve the problems of easy hemolysis, high production cost of antimicrobial peptides, low biological activity, etc., and achieve chemical synthesis Low difficulty, significant antibacterial effect, and low hemolytic activity

Active Publication Date: 2022-04-05
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although antimicrobial peptides have shown great potential in the treatment of bacterial infections, most of the current antimicrobial peptides still have problems such as high production costs, low biological activity, and easy hemolysis.

Method used

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  • A kind of low molecular weight antimicrobial peptide yhx-2 and its composition and application
  • A kind of low molecular weight antimicrobial peptide yhx-2 and its composition and application
  • A kind of low molecular weight antimicrobial peptide yhx-2 and its composition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The design of embodiment 1 antimicrobial peptide

[0028] 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, 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 shown in Table 1 below:

[0029] Table 1 Sequence parameters of de novo designed antimicrobial peptides

[0030] sequence length charged number Number of hydrophobic amino acids amino acid composition 13 +4 7 Gly, Lys, Ser, Leu

[0031] Considering factors such as hemoly...

Embodiment 2

[0044] Embodiment 2 antibacterial activity detection

[0045] 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%.

[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] (1) Inhibition zone test

[0048] Prepare LB and BHI semi-solid medium (mass fraction of agar 0.6%), add 7 μL of bacterial solution to each plate of 20mL, shake and mix well, then pour it into the petri dis...

Embodiment 3

[0054] Example 3 Detection of hemolytic activity

[0055] Take 1mL of fresh healthy rabbit blood and add it to a heparin anticoagulant tube, centrifuge at 1000xg for 10min, get the precipitate, wash it with PBS buffer 3 times, and resuspend the red blood cells in 10mL of PBS. Adjust the concentration of antimicrobial peptide YHX-2 to 4-512 μg / mL with PBS buffer, add it into a 96-well plate at a dose of 50 μL per well, 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.

[0056] The calculation formula of hemolysis rate is: hemolysis rate=(A T -A O ) / (A C -A O )×100%;

[0057] In the formula, A T is the absorbance value of the experimental group, A C is the ...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a low-molecular-weight antimicrobial peptide YHX‑2 and its composition and application. The amino acid sequence of the antimicrobial peptide YHX‑2 is shown in SEQ ID No.1. The antibacterial peptide has the characteristics of simple artificial synthesis, significant antibacterial effect and low hemolytic activity; it has broad-spectrum antibacterial activity, can inhibit Gram-positive bacteria and Gram-negative bacteria, and has high antibacterial activity; its The synthetic sequence is short, the molecular weight is small, and the chemical synthesis is less difficult. It can kill pathogenic bacteria in the organism more specifically and save the cost of large-scale production. It has broad application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a low-molecular-weight antimicrobial peptide YHX-2 and its composition and application. Background technique [0002] Since Fleming discovered penicillin, antibiotics have played an irreplaceable role in the fight between humans and pathogens, and have become the most commonly used class of drugs for humans to treat infectious diseases. However, due to the long-term and extensive use and abuse of antibiotics, the number of drug-resistant strains in clinical and animal husbandry has been increasing in recent years, and it has also led to the emergence of multi-drug resistant bacteria. At present, the common multidrug-resistant bacteria in clinical practice are: methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Pseudomonas aeruginosa (MDR-PA), multidrug-resistant Acinetobacter baumannii (MDR-AB) and vancomycin-resistant enterococci (VRE). At present,...

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 易华西李剑勋刘同杰公丕民张兰威郝彦玲步雨珊
Owner OCEAN UNIV OF CHINA
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