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Cow milk beta-lactoglobulin T cell immune tolerance hydrolysates as well as preparation method and application thereof

A technology of lactoglobulin and cellular immunity, applied in the direction of plasma globulin/lactoglobulin, mammalian protein, application, etc., can solve the problem of milk affecting the food safety of people with allergies, and achieve the effect of preventing allergic reactions in mice

Active Publication Date: 2021-08-20
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cow's milk allergy has seriously affected the food safety of those with allergies

Method used

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  • Cow milk beta-lactoglobulin T cell immune tolerance hydrolysates as well as preparation method and application thereof
  • Cow milk beta-lactoglobulin T cell immune tolerance hydrolysates as well as preparation method and application thereof
  • Cow milk beta-lactoglobulin T cell immune tolerance hydrolysates as well as preparation method and application thereof

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preparation example Construction

[0065] The method for preparing bovine milk β-lactoglobulin T cell immune tolerance hydrolyzate is characterized in that it comprises the following steps:

[0066] 1) preparing β-lactoglobulin into a 3% (w / v) aqueous solution;

[0067] 2) According to the ratio of 3000U / g protein, add protease to the aqueous solution of β-lactoglobulin to adjust the pH value of the reaction system; hydrolyze the reaction at a temperature of 50-60°C for 3-5h; shake the hydrolysis reaction in a constant temperature water bath performed in a machine; all examples

[0068] 3) Inactivate the enzyme in a water bath at 85°C for 10 minutes and cool down.

Embodiment 1

[0070] Bioinformatics prediction of β-lactoglobulin T cell epitopes:

[0071] 1) Materials and methods:

[0072] Get the amino acid sequence of lactoglobulin from GenBank, through IEDB (http: / / www.iedb.org / ), NetMHC(Ⅱ) / pan4.0 (https: / / services.healthtech.dtu.dk / service.php ?NetMHCIIpan-4.0) online site tool to predict T cell epitopes. On the IEDB homepage, first select the T cell epitope prediction section, enter the β-LG amino acid sequence or upload the Fasta file of the BLG sequence, select the pre-combined MHC molecule, and set the length of the polypeptide, and you will get the possible β predicted by the website -LG T cell epitope, and finally output the result. The smaller the adjustment level in the results, the more likely it is the β-lactoglobulin T cell epitope. On the NetMHC(II) / pan4.0 webpage, input the β-LG amino acid sequence or upload the Fasta file of the BLG sequence, select the pre-combined MHC molecular information, and set the length of the polypeptide,...

Embodiment 2

[0084] The method for preparing bovine milk β-lactoglobulin T cell immune tolerance hydrolyzate is characterized in that it comprises the following steps:

[0085] 1) preparing β-lactoglobulin into a 3% (w / v) aqueous solution;

[0086] 2) Add protease to the aqueous solution of β-lactoglobulin according to the ratio of 3000U / g protein, adjust the pH value of the reaction system, and carry out the hydrolysis reaction; the hydrolysis reaction is carried out in a constant temperature water bath shaker; all the protease hydrolysis reaction conditions in the embodiment as shown in Table 3;

[0087] 3) Inactivate the enzyme in a water bath at 85°C for 10 minutes and cool down.

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Abstract

The invention relates to a cow milk beta-lactoglobulin T cell immune tolerance hydrolysates as well as a preparation method and application thereof. The method comprises the following steps: predicting beta-lactoglobulin T cell epitopes; performing enzymolysis on beta-lactoglobulin by using different types of proteases, determining amino acid sequences of peptide fragments of hydrolysates by using a mass spectrum, comparing the amino acid sequences of the peptide fragments with predicted T cell epitope sequences of the beta-lactoglobulin, and preliminarily screening out the hydrolysates which possibly have an immune tolerance effect; and using the screened hydrolysates which possibly have the immune tolerance effect as research objects, and further screening the hydrolysates which really have the immune tolerance effect through animal experiments. According to the invention, models for inducing Balb / c mouse oral tolerance by trypsin, compound protease and papain beta-lactoglobulin hydrolysates are successfully established, and the three hydrolysates can effectively prevent OVA-induced mouse anaphylactic reaction. Peptide drugs or functional foods can be developed from the beta-lactoglobulin hydrolysates according to the present invention.

Description

technical field [0001] The invention relates to the field of food biotechnology, in particular to a bovine milk β-lactoglobulin T cell immune tolerance hydrolyzate and a preparation method and application thereof. Background technique [0002] With the continuous development of the food industry, food safety has received more and more attention. In recent years, the phenomenon of food allergy has been increasing year by year. According to a large number of studies, among food allergies, milk protein allergy is the most common. Milk is not only the best source of dietary protein for infants and young children, but also necessary for adults to obtain protein. It has excellent functional properties and provides rich nutrition for the body. However, cow's milk allergy has seriously affected the food safety of people with allergies. Nearly 20% of the total milk protein is whey protein, and α-lactalbumin and β-lactoglobulin are the two main whey proteins, accounting for 20% and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12P21/06A23L33/18A23J3/34A61K38/17
CPCC07K14/4717C12P21/06A23J3/343A23L33/18A61P37/08A23V2002/00A61K38/00A23V2250/55
Inventor 丛艳君张倩倩
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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