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Application of natural host defense peptide Alligatorin5

A host defense peptide, a natural technology, applied in the field of biomedicine, can solve the problems of bacterial drug resistance and unscientific drug residues, achieve broad-spectrum and high-efficiency antibacterial effects, improve resistance, and have wide application prospects

Active Publication Date: 2017-10-20
中科肽谷(山东)生命科学研究有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 Alligatorin5
  • Application of natural host defense peptide Alligatorin5
  • Application of natural host defense peptide Alligatorin5

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1 Preparation of Alligatorin5

[0019] (1) Using an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize the complete sequence of Alligatorin5 derived from reptiles, alligatorin 5, 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) Purified Alligatorin5 was identified by high performance liquid chromatography (HPLC) for its purity, its molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), its isoelectric point was determined by isoelectric focusing electrophoresis, and it was determined by an automatic amino acid sequencer Amino acid sequence structure.

[0022] The amino acid sequence of the natural host defense peptide Alligatorin5 of the present invention is shown in SEQ ID No.1. It consist...

Embodiment 2

[0023] Example 2 Detection of antibacterial activity of Alligatorin5 against 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 5 sample solution of 2 mg / ml dissolved in sterilized deionized water on the base surface, 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), Alligatorin5 minimum inhibitory concentration (Minimum Inhibitory Concentration) determination (2-fold dilution method):

[0026] Select the strains with...

Embodiment 3

[0034] Embodiment 3 Alligatorin5 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 Alligatorin5 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 Alligatorin5 samples in a 37°C incubator for shaking culture, take 50 μl of the bacterial solution at 0, 10, 20, 30, 45, 60, 90, 120 and 180 minutes to dilute 1000 times, 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 5 has a rapid b...

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Abstract

The invention relates to application of natural host defense peptide Alligatorin5 to preparation of aquatic pathogenic bacterium resisting medicine. The invention also discloses application of the natural host defense peptide Alligatorin5 to preparation of an aquaculture animal immune activating agent. The invention further discloses application of the natural host defense peptide Alligatorin5 to preparation of aquaculture animal feed. The results of study using hairy crabs as experimental animals show that the Alligatorin5 can enhance the immune function of the hairy crabs; the immunity of the hairy crabs on bacterial infection is improved; the survival rate of the hairy crabs is improved. The Alligatorin5 has wide 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 Alligatorin5. Background technique [0002] my country is a large breeding country. 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 excessive antibiotic...

Claims

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Application Information

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IPC IPC(8): A61K38/17A23K20/147A61P31/04
CPCA61K38/1709A23K20/147
Inventor 王义鹏于海宁郭志来陈燕冯婷婷
Owner 中科肽谷(山东)生命科学研究有限公司
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