Ovotransferrin antibacterial peptide and preparation method thereof

An ovotransferrin and antimicrobial peptide technology, which is applied in the field of bioactive peptide extraction, can solve the problems of sequencing identification, large molecular weight of antimicrobial active peptides, etc., and achieves the effect of high antimicrobial activity

Active Publication Date: 2021-11-09
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method does not sequence and identify the hydrolyzate with high antibacterial activity, and the molecular weight of the antibacterial active peptide obtained is relatively large

Method used

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  • Ovotransferrin antibacterial peptide and preparation method thereof
  • Ovotransferrin antibacterial peptide and preparation method thereof
  • Ovotransferrin antibacterial peptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) Pretreatment of ovotransferrin

[0036] Ovotransferrin was formulated as a 2% solution, heated in a water bath at 80°C for 20min with constant stirring, and then cooled to 35°C.

[0037] (2) Pepsin enzymolysis protein solution

[0038] The pH value of the protein solution was adjusted to 2, 1% pepsin was added, and the protein hydrolyzate was obtained by enzymatic hydrolysis at 37° C. for 100 min. After centrifugation at 10,000 g for 20 min, the supernatant was taken and freeze-dried in a freeze dryer.

[0039] (3) Weak cation column separation and purification

[0040] Freeze-dried ovotransferrin hydrolyzate was dissolved in 10 mM acetate buffer at pH 3. The sample solution was added to 150 system HiTrap CM FF ion exchange column. Equilibrate the column with 5 times the column volume of acetic acid buffer, and then carry out linear elution, wherein solution A is acetic acid buffer, solution B is acetic acid buffer containing 1M sodium chloride, and the sample ...

Embodiment 2

[0054] (1) Pretreatment of ovotransferrin

[0055] Ovotransferrin was formulated as a 2% solution, heated in a water bath at 80°C for 20min with constant stirring, and then cooled to 35°C.

[0056] (2) Pepsin enzymolysis protein solution

[0057] The pH value of the protein solution was adjusted to 2, 1% pepsin was added, and the protein hydrolyzate was obtained by enzymatic hydrolysis at 37° C. for 100 min. After centrifugation at 10,000 g for 20 min, the supernatant was lyophilized in a freeze dryer.

[0058] (3) Weak cation column separation and purification

[0059] Freeze-dried ovotransferrin hydrolyzate was dissolved in 10 mM acetate buffer at pH 3. The sample solution was added to 150 system HiTrap CM FF ion exchange column. Equilibrate the column with 5 times the column volume of acetic acid buffer, and then carry out linear elution, wherein solution A is acetic acid buffer, solution B is acetic acid buffer containing 1M sodium chloride, and the sample is eluted a...

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Abstract

The invention provides an ovotransferrin antibacterial peptide and a preparation method thereof. In this method, ovotransferrin is hydrolyzed by pepsin after heat treatment, separated, purified and identified to obtain three highly active antibacterial peptides. The present invention pretreats the protein by heating, which lays a good substrate foundation for pepsin hydrolysis to obtain highly active small molecule antibacterial peptides; at the same time, it adopts weak cation chromatographic column and reverse liquid chromatographic column for separation and purification, which is beneficial to obtain The peptide sequence with high antibacterial activity can be used in food industry and medical fields.

Description

technical field [0001] The invention belongs to the technical field of bioactive peptide extraction, and in particular relates to an ovotransferrin antibacterial peptide and a preparation method thereof. Background technique [0002] Eggs are an important source of biologically active protein, which contains ovalbumin, ovotransferrin, ovomucin, ovoglobulin and lysozyme. Ovotransferrin has a molecular weight of 76 kDa, has 686 amino acid residues and is an iron-binding glycoprotein that accounts for 12% of egg white protein. It consists of two globular leaflets (N and C leaflets) of similar size, each divided into two domains (domains N1 and N2 in the N-lobe and C1 and C2 domains in the C-lobe). In recent years, various physiological benefits of ovotransferrin have been reported, such as antibacterial, immunomodulatory, anticancer, antiviral and antioxidant properties, and angiotensin converting enzyme inhibitory activity. Ovotransferrin is degraded into peptides during dig...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/79C12P21/06C07K1/20C07K1/18
CPCC07K14/79C12P21/06
Inventor 金永国付星马斌马美湖
Owner HUAZHONG AGRI UNIV
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