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Preparation method of goat cheese protein source antibacterial peptide

A technology of goat cheese and antibacterial peptides, which is applied in the fields of peptide preparation methods, antibacterial drugs, chemical instruments and methods, etc., can solve the problems of unknown primary and secondary structures of antibacterial peptides, achieve excellent antibacterial properties and broad application prospects Effect

Inactive Publication Date: 2014-09-10
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented process allows us to create an anti-microorganism substance called geckosanoprotein (GPC). These small molecules can be found naturally or added during processing steps like baking. They also provide important benefits such as being effective at killing harmful microbes without causing any damage on their owners' healthy tissues.

Problems solved by technology

Technologies related to biomarkers involved in identifying diseases caused by increased levels of inflammatory cytokines like TNFα, IL-1β, IFNa-2, PARP-4, MMP-3, LPS, FMLM, SMRT, SIRT, DNA helix stabilizers, ribonucleotides, nucleated polypeptides containing lanthanum cations, carboxylate groups, metal ion binding sites, and nonionic charges. These agents help control harmful metabolites released during stress responses associated with these processes. However, they may cause unwanted side effects when taken up into living cells due to low solubility and instability issues. Therefore, finding ways to improve the biocompatibility of existing medicinal products while maintaining its efficacy remains an issue.

Method used

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  • Preparation method of goat cheese protein source antibacterial peptide
  • Preparation method of goat cheese protein source antibacterial peptide
  • Preparation method of goat cheese protein source antibacterial peptide

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preparation example Construction

[0031] 2) Preparation of casein hydrolysate

[0032] Weigh an appropriate amount of crude goat cheese protein and place it in an enzymatic hydrolysis tank, use 0.1mol / L sodium hydroxide solution to aid the solution, and dilute to the specified volume with deionized water to obtain 30g / kg~120g / kg (optimized to 60g / kg~100g / kg) goat cheese protein solution, adjust the pH to 5.0~7.0 (optimize 5.5~6.5), heat at 95℃ for 5min, cool to 50℃~70℃, add 3000U / g~7000U / g( Optimize the papain of 4000U / g~6000U / g, add 2mol / L NaOH solution during the hydrolysis process to keep the pH constant. After enzymatic hydrolysis for 0.5h~4.0h (optimization 1.0h~2.0h), kill at 95℃ Enzyme for 10 minutes, cooled to room temperature, centrifuged at 10000×g for 15 minutes, and the supernatant was taken and concentrated at 45-50°C and 0.09MPa to obtain the goat cheese proteolysis concentrate (i.e. crude antimicrobial peptide).

[0033] 3) Desalination and separation and purification of antimicrobial peptides

[00...

Embodiment 1

[0055] (1) Take fresh goat milk to filter and remove impurities, centrifuge at 5000r / min at 4℃ for 20min to remove milk fat, adjust the pH value of skimmed goat milk to 4.3 with 2mol / L HCl solution, and let it stand in a water bath at 40℃ for 30min to precipitate casein. Centrifuge at 4000r / min for 10min, discard the supernatant, wash the pellet twice with pH 4.3 acetic acid-sodium acetate buffer, and centrifuge at 4000r / min for 10 min after each wash, the resulting precipitate is crude casein, Kjeldahl Nitrogen method determines the protein content of crude casein.

[0056] (2) Weigh a certain amount of goat milk crude casein and place it in the enzymolysis tank, use 0.1 mol / L sodium hydroxide solution to aid the solution, and dilute to the specified volume with deionized water to obtain 80 g / kg of goat cheese For protein solution, adjust the pH to 6.0, heat at 95°C for 5 minutes, cool to 60°C, add 5000U / g papain accurately, and add 2mol / L NaOH solution during hydrolysis to keep...

Embodiment 2

[0061] 1) Take fresh goat milk to filter and remove impurities, centrifuge at low temperature to defat, adjust the pH value of skim goat milk to 4.3 with 2mol / L HCl solution, let stand at 40℃ in water bath for 30 min to precipitate casein, centrifuge at 4000r / min for 10 min, discard the supernatant The precipitate was washed twice with pH 4.3 acetic acid-sodium acetate buffer, and centrifuged at 4000 r / min for 10 minutes after each washing, and the obtained precipitate was goat milk crude casein.

[0062] (2) Weigh a certain amount of goat milk crude casein and place it in an enzymatic hydrolysis tank, use 0.1 mol / L sodium hydroxide solution to aid the solution, and dilute to the specified volume with deionized water to obtain 70 g / kg goat For the casein solution, adjust the pH to 6.0, heat at 95°C for 5 minutes, cool to 60°C, add 6000U / g papain accurately, add 2mol / L NaOH solution during the hydrolysis process to keep the pH at 6.0±0.1, 60 After enzymatic hydrolysis at ℃ for 80 ...

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Abstract

The invention discloses a preparation method of goat cheese protein source antibacterial peptide. The preparation method comprises the following steps: performing enzymolysis on the goat cheese protein by use of papain; separating and desalting the zymolyte by use of macroporous nonpolar resin; purifying by semi-preparation gel permeation chromatograph; and performing freeze drying to obtain goat cheese protein source antibacterial peptide powder. The result of the trial experiment on antibacterial function carried out by the applicant indicates that the molecular weight of the prepared goat cheese protein source antibacterial peptide is below 3,000, and the goat cheese protein source antibacterial peptide is easy to absorb and utilize; the secondary structure of the obtained goat cheese protein source antibacterial peptide is mainly a random coil; and the goat cheese protein source antibacterial peptide is a nontoxic green natural antibacterial peptide, has good antibacterial properties on escherichia coli, staphylococcus aureus, salmonella and the like, and has a remarkably broad application prospect in the preservation of milk and dairy products, cosmetics and other food.

Description

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Claims

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Application Information

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Owner NORTHWEST A & F UNIV
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