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A kind of antimicrobial peptide and its application

A technology of antimicrobial peptide and antibacterial concentration, which is applied in the field of antimicrobial peptide and its application, can solve the problems of low biological activity and high production cost, and achieve the effect of low hemolytic activity, short synthetic sequence and high antibacterial activity

Active Publication Date: 2022-05-27
YANGZHOU YANGDA KANGYUAN DAIRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most antimicrobial peptides still have problems such as high production cost and low biological activity.

Method used

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  • A kind of antimicrobial peptide and its application
  • A kind of antimicrobial peptide and its application
  • A kind of antimicrobial peptide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Design of antimicrobial peptides

[0030] 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 bacteriostatic effects on Gram-positive and Gram-negative bacteria were analyzed (including sequence length, number of charges, hydrophobic amino acid ratio and amino acid composition), and then according to the selection principle of the largest frequency of occurrence and combined with rational design ideas, the sequence parameters of the new antimicrobial peptides were determined, as follows:

[0031] Table 1 Sequence parameters for de novo design of antimicrobial peptides

[0032]

[0033] In order to reduce the cost of synthesis while reducing cytotoxicity, the sequence length of 13 was chosen as the second most frequent. The positively charged amino acid Lys and polar uncharged amino acid Ser were...

Embodiment 2

[0044] Example 2 Antibacterial activity

[0045] Staphylococcus aureus, Escherichia coli and Salmonella were streaked on LB solid medium (Luria-Bertani medium), and Listeria monocytogenes and Streptococcus mutans were streaked on BHI solid medium (brain). Heart infusion agar medium), placed in a constant temperature incubator at 37°C for 18h, picking a single colony of each strain and placing it in the corresponding liquid medium, and incubated at 37°C with constant temperature shaking for 12h. Measure the OD of bacterial liquid 600 value (the absorbance value of the solution at 600nm wavelength) and diluted to 1×10 6 CFU / mL.

[0046] ①Inhibition zone experiment

[0047] Configure LB and BHI semi-solid medium (agar mass fraction of 0.6%), add 7 μL of bacterial liquid to 20 mL of each dish, shake and mix, and pour it into the Petri dish placed in the Oxford cup. After the medium cools and solidifies, remove the Oxford cup to complete the mixing. hole. Add 160 μL of antimic...

Embodiment 3

[0053] Example 3 Hemolytic activity

[0054] 1 mL of healthy rabbit blood was added to a heparin anticoagulant tube, centrifuged at 1000×g for 10 min, and the pellet was collected, washed three times with PBS buffer, and the red blood cells were resuspended in 10 mL of PBS. The concentration of antimicrobial peptide YHX-1 was adjusted to 4 μg / mL, 8 μg / mL, 16 μg / mL, 32 μg / mL, 128 μg / mL, 256 μg / mL, 512 μg / mL with PBS buffer, and an equal volume of red blood cell suspension was added. Take PBS buffer as a negative control and 0.1% Tritonx-100 (polyethylene glycol octyl phenyl ether) as a negative control, incubate at 37°C for 1 h, take out, centrifuge at 1000 x g for 10 min, take out the supernatant and use a microplate reader OD value was measured at 570nm.

[0055] The calculation formula of hemolysis rate is: hemolysis rate = (A T -A 0 ) / (A C -A 0 ) × 100%.

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

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Abstract

The invention discloses an antibacterial peptide and application thereof. The antibacterial peptide is antibacterial peptide YHX‑1 having the amino acid sequence shown in SEQ ID NO.1. The antimicrobial peptide has broad-spectrum antibacterial activity, can inhibit Gram-positive bacteria such as Listeria monocytogenes, Staphylococcus aureus, and Streptococcus mutans, and Gram-negative bacteria such as Escherichia coli and Salmonella, and all have High antibacterial activity. In addition, antimicrobial peptides have low hemolytic activity and low toxicity, and the synthetic sequence is short, the molecular weight is small, and the chemical synthesis is not difficult. It can kill pathogenic bacteria in the organism with good specificity while ensuring safety. In addition, it can also save scale. minimize production costs.

Description

technical field [0001] The present invention relates to antibacterial agents, in particular, to an antibacterial peptide and its application. Background technique [0002] Since the advent of the world's first antibiotic, penicillin, antibiotics have played an irreplaceable role in the fight against pathogens. However, in recent years, the abuse of antibiotics has led to the continuous increase of drug-resistant strains and the emergence of multi-drug-resistant bacteria, posing a serious threat to the treatment of clinical infections. For example, Methicillin-resistant Staphylococcusaureus (MRSA) has broad-spectrum drug resistance and is resistant to all β-lactam and cephalosporin antibiotics similar in structure to methicillin, as well as aminoglycosides, Macrolides, tetracyclines, fluoroquinolones, sulfonamides, and rifampicin have different degrees of resistance. MRSA is also a common pathogen of nosocomial infection, and its infection is more common in trauma and surge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A23L3/3526A23K20/147C11D3/48C11D3/32C11D7/32C11D9/30A61K38/10A61P31/04A61K8/64A61Q19/00
CPCC07K7/08A23L3/3526A23K20/147C11D3/48C11D3/32C11D7/3263C11D9/30A61P31/04A61K8/64A61Q19/00A61K38/00A61K2800/10A61K2800/524A23V2002/00A23V2200/10A23V2250/55Y02A50/30
Inventor 易华西李剑勋周炜印伯星杨仁琴徐广新杨忠良王锦荣吴长清刘阳张海霞吴连萍贾若微李沐蓉陈佳顾文静华惠胡潇张忆梅
Owner YANGZHOU YANGDA KANGYUAN DAIRY
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