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Application of natural host defense peptide alligatorin4

A host defense peptide, a natural technology, applied in the field of biomedicine, can solve the problems of unscientific, bacterial drug-resistant drug residues, etc., and achieve the effect of improving resistance, broad-spectrum and high-efficiency antibacterial effect, and improving survival rate

Active Publication Date: 2019-12-03
思格(苏州)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, with the extensive use of antibiotics in the aquaculture industry, especially the unscientific abuse, problems such as bacterial resistance and drug residues have become increasingly prominent

Method used

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  • Application of natural host defense peptide alligatorin4
  • Application of natural host defense peptide alligatorin4
  • Application of natural host defense peptide alligatorin4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of Alligatorin4

[0019] (1) Using an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize the full sequence of Alligatorin4 derived from reptiles, alligatorin 4, and desalting and purifying it by HPLC reverse-phase column chromatography.

[0020] (2) The molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0021] (3), the purified Alligatorin4 is identified by high performance liquid chromatography (HPLC) for its purity, the molecular weight is determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), the isoelectric point is determined by isoelectric focusing electrophoresis, and it is determined by an automatic amino acid sequencer Amino acid sequence structure.

[0022] The amino acid sequence of the natural host defense peptide Alligatorin4 of the present invention is shown in SEQ ID No.1. It consists of 3...

Embodiment 2

[0023] Example 2 Detection of antibacterial activity of Alligatorin4 on common aquatic pathogens

[0024] (1), the test strains preserved on the slant were picked and evenly spread on the nutrient broth solid medium (nutrient broth medium, British OXOID company) plate, and the sterilized filter paper sheet with a diameter of 0.5cm was placed in the culture medium. Add dropwise 10 μl of Alligatorin 4 sample solution of 2 mg / ml dissolved in sterilized deionized water on the surface of the base, and incubate upside down at 37°C for 18-20 hours to observe whether the inhibition zone is formed or not. If the sample has antibacterial activity, a clear and transparent bacteriostatic zone will be formed around the filter paper, and the larger the bacteriostatic zone, the stronger the antibacterial activity of the sample.

[0025] (2), Alligatorin4 minimum inhibitory concentration (Minimum Inhibitory Concentration) determination (2-fold dilution method):

[0026] Select the strains wi...

Embodiment 3

[0034] Embodiment 3 Alligatorin4 sterilization speed measurement

[0035] Vibrio parahaemolyticus was cultured at 37°C for 12 hours with NB liquid medium (OXOID, UK), and then diluted to 10 with fresh NB liquid medium. 6 CFU / ml bacterial suspension. The Alligatorin4 sample dissolved in sterilized deionized water was added to the bacterial suspension so that the final concentration was 5×MIC (23.45 μg / ml). Place the bacterial solution added with Alligatorin4 sample in a 37°C incubator for shaking culture, take 50 μl of the bacterial solution to dilute 1000 times at 0, 10, 20, 30, 45, 60, 90, 120 and 180 minutes, and then take 50 μl of the diluted The bacterial solution was spread on the NB solid medium plate, and the colonies were counted after culturing overnight in a 37°C incubator. Ampicillin was used as a positive control in this experiment, and sterile deionized water was used as a negative control.

[0036] The results are shown in Table 3. Alligatorin 4 has a rapid ba...

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Abstract

The invention relates to application of natural host defense peptides Alligatorin4 to preparing aquatic pathogenic bacterium resistant medicines, aquaculture animal immunity activators and aquaculture animal feed. The application has the advantages that as shown by results of research on Chinese mitten crab which is an experimental animal, immunity functions of the Chinese mitten crab can be enhanced by the natural host defense peptides Alligatorin4, the bacterial infection resistance of the Chinese mitten crab can be improved, and the survival rate of the Chinese mitten crab can be increased; the natural host defense peptides Alligatorin4 have broad application prospects in the field of aquaculture.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a natural host defense peptide Alligatorin4. Background technique [0002] my country is a big country in the breeding industry. The total amount of poultry eggs, meat and aquatic products has ranked first in the world for many years, and the total output value has reached more than 1 trillion yuan. Antibiotics are chemicals produced by microorganisms that, at low concentrations, inhibit or kill other microorganisms. Antibiotics have played an important role in the development of aquaculture because of their functions of promoting animal growth, improving feed utilization, and preventing and treating animal diseases. [0003] However, with the extensive use of antibiotics in the aquaculture industry, especially the unscientific abuse, problems such as bacterial resistance and drug residues have become increasingly prominent. Due to problems such as excess...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P31/04A61P37/04A23K20/195A23K20/147
CPCA23K20/147A23K20/195A61K38/1703Y02A50/30
Inventor 王义鹏于海宁冯婷婷郭志来陈燕
Owner 思格(苏州)生物科技有限公司
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