Hybrid antibacterial peptide based on fv7 anti-biofilm and its preparation method and application

A hybrid antimicrobial peptide and anti-biofilm technology, applied in the field of agricultural, animal husbandry and veterinary medicine, can solve the problems of less application in the field of medicine and animal husbandry, low antibacterial activity, etc., and achieve the goal of improving cell selectivity, enhancing antibacterial activity, and reducing costs Effect

Active Publication Date: 2018-11-20
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are many kinds of antimicrobial peptides in nature and they are widely distributed, but they are rarely used in the fields of medicine and animal husbandry
There are many factors that limit the application of antimicrobial peptides, and for some antibacterial peptides, low antibacterial activity is the main factor limiting their application

Method used

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  • Hybrid antibacterial peptide based on fv7 anti-biofilm and its preparation method and application
  • Hybrid antibacterial peptide based on fv7 anti-biofilm and its preparation method and application
  • Hybrid antibacterial peptide based on fv7 anti-biofilm and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Design and synthesis of antimicrobial peptides

[0016] LL: The 17th-29th amino acid functional sequence of the human antimicrobial peptide LL-37, this fragment is a peptide chain with an amphipathic helical structure rich in cations. The shortest sequence identified as having antibacterial and anticancer activities can effectively bind to anionic phospholipids of bacterial cell membranes and exhibit selective cytotoxicity, but no toxicity to human cells.

[0017] A sequence FV7 with anti-biofilm activity and a polypeptide fragment LL are hybridized to obtain a hybrid antibacterial peptide FV-LL. The helical wheel prediction diagram of the hybrid peptide FV-LL is as follows figure 1 shown. Using a peptide synthesizer, the above three antimicrobial peptides were synthesized by solid-phase synthesis. The sequences and physicochemical parameters of the three peptides are shown in Table 1.

[0018] Table 1 The sequences and physicochemical parameters of the th...

Embodiment 2

[0021] The physical and chemical index analysis of embodiment 2 antimicrobial peptides

[0022] The secondary structure of the antimicrobial peptide was analyzed by CD spectrum, and the results showed that both the hybrid peptide and the original peptide showed a random coil structure in the water environment, while in the hydrophobic environment (50% TFE) of the simulated microbial membrane and negative In the charged prokaryotic membrane environment (30mM SDS), both the original peptide LL and the hybrid peptide FV-LL showed a certain α-helical structure, while FV7 showed a certain β-sheet conformation.

[0023] The stability analysis of antimicrobial peptides showed that the hybrid peptide FV-LL had ideal resistance to heat treatment. However, the antibacterial activity of the peptide FV-LL decreased slightly at different physiological concentrations of salt ions.

Embodiment 3

[0024] Bacteriostasis and anti-biofilm activity of embodiment 3 antimicrobial peptides

[0025] 1 Antibacterial activity

[0026] The peptide was configured as a 2.56mM / L storage solution for future use. The minimum inhibitory concentrations of several antimicrobial peptides were determined by the broth microdilution method. Using 0.01% acetic acid (containing 0.2% BSA) as the diluent, a series of gradient antimicrobial peptide solutions were sequentially prepared using the double dilution method. Take 100 μL of the above solution and place it in a 96-well cell culture plate, then add an equal volume of the bacteria solution to be tested (~10 5 individual / mL) in each well. Positive controls (containing bacterial fluid but not antimicrobial peptides) and negative controls (neither bacterial fluid nor peptides) were set up. Incubate at a constant temperature of 37°C for 20 hours, and the minimum inhibitory concentration is the one where no turbidity is seen at the bottom of ...

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Abstract

The invention relates to a hybrid antibacterial peptide based on an FV7 anti-bio-membrane, and a preparation method and an application thereof. The sequence of the hybrid antibacterial peptide is represented as the SEQ ID No.1. According to main physical and chemical parameters and structural and functional relationship of an original peptide and a hybrid peptide, an anti-bio-membrane fragment FV7 and an active centre fragment LL of a natural antibacterial peptide LL-37 are synthesized through a chemical method to obtain the hybrid peptide FV-LL. Through a series of in-vitro test, the antibacterial activities of the original peptides FV7 and LL and the hybrid antibacterial peptide FV-LL are verified, and through comparison, it is found that the FV-LL is greatly improved in antibacterial activity and also has anti-bio-membrane activity, so that through functional fragment hybridization, the antibacterial activity of the antibacterial peptide can be improved, thereby improving cell selectivity. Furthermore, the antibacterial mechanism of the hybrid antibacterial peptide is verified, thereby providing theoretical basis for application of the antibacterial peptide.

Description

technical field [0001] The invention belongs to the application field of agriculture, animal husbandry and veterinary medicine, and specifically relates to a hybrid antibacterial peptide based on FV7 anti-biofilm, its preparation method and application. Background technique [0002] The abuse of feed antibiotics leads to environmental pollution, drug residues and bacterial resistance, which has brought great harm to agriculture and animal husbandry, seriously affecting the maximum potential of animal production and the green and safe livestock and poultry products. supply. Therefore, finding novel, residue-free and highly efficient immunocatalysts that can replace antibiotics has become an urgent problem to be solved. As a kind of small molecular polypeptide for animal body to defend against external microorganisms, antimicrobial peptides have the advantages of broad-spectrum antibacterial properties, no drug resistance, no toxicity, no pollution and no residue, etc., and g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C07K1/04A61K38/10A61P31/04
CPCA61K38/00C07K14/00C07K14/47C07K2319/00
Inventor 单安山谭婷婷吴迪
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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