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Antibacterial peptide and application thereof

An antibacterial peptide and antibacterial concentration technology, applied to the antibacterial peptide and its application field, can solve the problems of high production cost and low biological activity, and achieve the effects of short synthetic sequence, low hemolytic activity, and large-scale production cost saving.

Active Publication Date: 2021-06-11
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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  • Antibacterial peptide and application thereof
  • Antibacterial peptide and application thereof
  • Antibacterial peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The design of embodiment 1 antimicrobial peptide

[0030] Based on the de novo design of antimicrobial peptides and the understanding of the structure-activity relationship, the sequence parameters of 843 antibacterial 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 selection principle of the highest frequency of occurrence and combined with rational design ideas, the sequence parameters of the new antimicrobial peptide were determined, as follows:

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

[0032]

[0033] In order to reduce the synthesis cost and reduce the cytotoxicity at the same time, the sequence length was selected as 13 which is the second most frequently used. The positively charged amino acid Lys and the pol...

Embodiment 2

[0044] Embodiment 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 18 hours, picked a single colony of each strain and placed them 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 (the absorbance value of the solution at a wavelength of 600nm), and dilute it to 1×10 6 CFU / mL.

[0046] ①Inhibition 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 Oxford cup placed, and pull out the Oxford cup after the medium is cooled and solidified to complete the infu...

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-1 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 add an equal volume of red blood cell suspension. Use PBS buffer as a negative control, and 0.1% Tritonx-100 (polyethylene glycol octylphenyl ether) 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 use a microplate reader Measure the OD value 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 the positive control grou...

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Abstract

The invention discloses an antibacterial peptide and application thereof. The antibacterial peptide is an antibacterial peptide YHX-1 with an amino acid sequence as shown in SEQ ID NO. 1. The antibacterial 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 has high antibacterial activity. In addition, the antibacterial peptide is relatively low in hemolytic activity, relatively low in toxicity, short in synthetic sequence, small in molecular weight and low in chemical synthesis difficulty, the safety is ensured while pathogenic bacteria in a living body can be well and specifically killed, and in addition, the large-scale production cost can also be saved.

Description

technical field [0001] The present invention relates to an antibacterial agent, in particular to an antibacterial peptide and its application. Background technique [0002] Since the advent of penicillin, the world's first antibiotic, antibiotics have been playing an irreplaceable role in the struggle between humans and 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, which pose a serious threat to the treatment of clinical infections. For example, Methicillin-resistant Staphylococcusaureus (MRSA) has broad-spectrum resistance to all β-lactam and cephalosporin antibiotics with similar structures to methicillin, as well as aminoglycosides, Macrolides, tetracyclines, fluoroquinolones, sulfonamides, and rifampan produced varying degrees of drug resistance. MRSA is also a common pathogen in nosocomial infections in hospitals. Its infection is more commo...

Claims

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Application Information

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Patent Type & Authority Applications(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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