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Method for extracting antimicrobial peptide from chicken blood

A technology of antimicrobial peptide and chicken blood, applied in the field of biological extraction, can solve the problems of low enzymatic hydrolysis efficiency, achieve the effect of improving enzymatic hydrolysis degree, low investment, and inhibiting thrombin activity

Inactive Publication Date: 2018-09-07
金华市铁骑士生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the chicken blood antibacterial agent is not treated with anticoagulant before enzymolysis, and the enzymolysis efficiency is low

Method used

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  • Method for extracting antimicrobial peptide from chicken blood

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Effect test

Embodiment 1

[0019] The method of extracting bacteriostatic peptides from chicken blood is to treat fresh chicken blood with a sodium hexametaphosphate aqueous solution containing fatty acid methyl ester and triethylamine for anticoagulation, then prepare a fermentation medium, inoculate Aspergillus niger to ferment the fermentation broth, and ferment The solution is subjected to ultrafiltration and gel column chromatography to obtain chicken blood bacteriostatic peptide. Aspergillus niger has a powerful polymer degrading enzyme system, which can grow and ferment quickly on a variety of cheap media. During the process of growth and reproduction, various metabolic enzymes, such as acid protease and neutral protease, are produced, which can be effective The degradation of macromolecular proteins into small molecular peptides has the advantages of low investment, simple operation, good protein degradation effect, and yield of antimicrobial peptides.

[0020] The method of extracting bacteriostat...

Embodiment 2

[0029] The method of extracting bacteriostatic peptide from chicken blood, its specific steps are:

[0030] 1) Anticoagulant treatment of chicken blood: add a 3% sodium hexametaphosphate aqueous solution to fresh chicken blood at a volume ratio of 1:5, stir evenly, and then centrifuge at a temperature of 4℃ and a speed of 5000r / min Centrifuge in the machine for 30 minutes, discard the precipitate, and freeze-dry the supernatant to obtain anticoagulated chicken blood meal for use;

[0031] 2) Preparation of fermentation medium: take 8 parts by weight of anticoagulant chicken blood meal, 20 parts of sucrose, 5 parts of sodium chloride, 0.005 parts of zinc chloride, 2.0 parts of potassium dihydrogen phosphate, 0.4 parts of magnesium sulfate, and 0.01 parts of iron sulfate Vitamin B 1 0.001 parts, 1000 parts of sterile water, mix evenly to obtain fermentation medium;

[0032] 3) Aspergillus niger fermentation: adjust the pH of the fermentation medium to 6.0, then inoculate Aspergillus n...

Embodiment 3

[0038] The method of extracting bacteriostatic peptide from chicken blood, its specific steps are:

[0039] 1) Anticoagulant treatment of chicken blood: add a 3% sodium hexametaphosphate aqueous solution to fresh chicken blood at a volume ratio of 1:5, stir evenly, and then centrifuge at a temperature of 4℃ and a speed of 5000r / min Centrifuge in the machine for 30 minutes, discard the precipitate, and freeze-dry the supernatant to obtain anticoagulated chicken blood meal for use;

[0040] 2) Preparation of fermentation medium: take 8 parts by weight of anticoagulant chicken blood meal, 20 parts of sucrose, 5 parts of sodium chloride, 0.005 parts of zinc chloride, 2.0 parts of potassium dihydrogen phosphate, 0.4 parts of magnesium sulfate, and 0.01 parts of iron sulfate Vitamin B 1 0.001 parts, 1000 parts of sterile water, mix evenly to obtain fermentation medium;

[0041] 3) Aspergillus niger fermentation: adjust the pH of the fermentation medium to 6.0, then inoculate Aspergillus n...

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Abstract

The invention discloses a method for extracting antimicrobial peptide from chicken blood. The method comprises the following specific steps: adding a sodium hexametaphosphate aqueous solution to freshchicken blood, performing stirring uniformly, performing centrifugation, then discarding precipitate, and adjusting pH of a supernatant to obtain anticoagulated chicken blood; preparing a fermentation medium from anticoagulated chicken blood powder, inoculating Aspergillus niger for fermentation, and centrifuging a supernatant to obtain a fermentation liquid; ultra-filtering the fermentation liquid to obtain crude antimicrobial peptide extract; performing gel chromatography on the crude antimicrobial peptide extract, performing elution with double distilled water after sample loading, collecting components with higher antimicrobial activity, and freeze-drying the components to obtain chicken blood antimicrobial peptide. The method for extracting antimicrobial peptide from the chicken blood is simple and feasible and has the beneficial effects that extraction efficiency, yield and purity are high, and the prepared antimicrobial peptide has high activity, can quickly kill bacteria andhas broad-spectrum antimicrobial properties.

Description

Technical field [0001] The present invention relates to the technical field of biological extraction, in particular to a method for extracting bacteriostatic peptides from chicken blood. Background technique [0002] Since the discovery of antibiotics, it has been widely used in medical treatment, traditional veterinary medicine and feed additives due to its good bacteriostatic effect. However, due to the excessive use of antibiotics, the drug resistance of pathogenic bacteria and drug residues have become more and more problems. The more serious it is, it has gradually attracted people's attention. It is urgent to find effective alternatives to antibiotics and solve the problems of drug resistance and drug residues. Antimicrobial peptides (AMPs) are a kind of peptides that can be produced by both prokaryotes and eukaryotes, which are widely distributed and have a wide range of antibacterial activities. They are resistant to gram-positive bacteria, gram Negative bacteria, fungi,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C07K1/36C07K1/34C07K1/16C12R1/685
CPCC12P21/00C07K1/16C07K1/34C07K1/36
Inventor 全盈园
Owner 金华市铁骑士生物科技有限公司
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