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 drug residues, unscientific and other problems, achieve broad-spectrum and high-efficiency antibacterial effects, improve resistance, and enhance immune function.

Active Publication Date: 2020-12-29
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

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 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]Example 1 Preparation of Alligatorin5

[0019](1) Use an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize the complete sequence of Alligatorin5 from the reptile alligator Alligatorin5, and purify it by HPLC reversed-phase column chromatography.

[0020](2) The determination of molecular weight adopts matrix-assisted laser analysis and ionization time-of-flight mass spectrometry (MALDI-TOF).

[0021](3) The purity of the purified Alligatorin 5 was identified by high performance liquid chromatography HPLC. The molecular weight was determined by matrix-assisted laser analysis and ionization time-of-flight mass spectrometry (MALDI-TOF), isoelectric focusing electrophoresis was used to determine the isoelectric point, and the automatic amino acid sequencer was used. 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 is composed of 22 amino acids, has a molecu...

Embodiment 2

[0023]Example 2 Detection of antibacterial activity of Alligatorin5 on common aquatic pathogens

[0024](1) Separately pick the test strains stored on the inclined plane and evenly spread them on the nutrient broth solid medium (nutrient broth medium, British OXOID company) plate, and place the sterilized 0.5 cm diameter filter paper sheet in the culture On the substrate surface, add 10 μl of a 2 mg / ml Alligatorin 5 sample solution dissolved in sterile deionized water, and incubate it upside down at 37°C for 18-20 hours to observe whether the inhibition zone is formed. If the sample has antibacterial activity, a clear and transparent zone of inhibition will be formed around the filter paper. The larger the zone of inhibition, the stronger the antibacterial activity of the sample.

[0025](2) Determination of Alligatorin5 Minimum Inhibitory Concentration (2-fold dilution method):

[0026]Select the strain with inhibition zone in the previous experiment for MIC determination experiment. The te...

Embodiment 3

[0034]Example 3 Determination of Alligatorin5 Sterilization Speed

[0035]Vibrio parahaemolyticus was cultured in NB liquid medium (OXOID, UK) at 37°C for 12 hours, and then diluted to 10 with fresh NB liquid medium6CFU / ml bacterial suspension. The Alligatorin5 sample dissolved in sterilized deionized water was added to the bacterial suspension to make the final concentration 5×MIC (23.45 μg / ml). Place the bacterial solution added with Alligatorin5 sample in a 37°C incubator with shaking culture. Dilute 50 μl bacterial solution 1000 times at 0, 10, 20, 30, 45, 60, 90, 120 and 180 minutes, and then take 50 μl 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. In this experiment, ampicillin was used as a positive control, and sterilized deionized water was used as a negative control.

[0036]The results are shown in Table 3. Alligatorin5 has a rapid sterilization rate for Vibrio parahaemolyticu...

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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 present invention relates to the field of biomedical technology, in particular to the application of a natural host defense peptide Alligatorin5.Background technique[0002]my country is a major producer of aquaculture. The total amount of poultry eggs, meat and aquatic products has ranked first in the world for many years, with a total output value of more than 1 trillion yuan. Antibiotics refer to chemical substances produced by microorganisms that can inhibit or kill other microorganisms at low concentrations. Antibiotics have played an important role in the development of the breeding industry because of their functions in promoting animal growth, improving feed utilization, preventing and treating animal diseases.[0003]However, with the extensive use of antibiotics in the breeding industry, especially unscientific abuse, the problems of 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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Patent Type & Authority Patents(China)
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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