A kind of milk β-lactoglobulin T cell immune tolerance hydrolyzate and its preparation method and application

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 allergic people, and achieve the effect of preventing allergic reactions in mice

Active Publication Date: 2022-07-19
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • 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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  • A kind of milk β-lactoglobulin T cell immune tolerance hydrolyzate and its preparation method and application
  • A kind of milk β-lactoglobulin T cell immune tolerance hydrolyzate and its preparation method and application
  • A kind of milk β-lactoglobulin T cell immune tolerance hydrolyzate and its preparation method and application

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preparation example Construction

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

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

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

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

Embodiment 1

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

[0071] 1) Materials and methods:

[0072] The amino acid sequence of lactoglobulin was obtained from GenBank, through IEDB (http: / / www.iedb.org / ), NetMHC(II) / pan4.0 (https: / / services.healthtech.dtu.dk / service.php ?NetMHCIIpan-4.0) online website 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-bound MHC molecule, and set the length of the peptide, and the possible β predicted by the website will be obtained. - LG T cell epitopes, and finally output the results. The smaller the adjustment level in the result, 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-bound MHC molecule information, set the length of the peptide...

Embodiment 2

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

[0085] 1) β-lactoglobulin is formulated 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 oscillator; all the protease hydrolysis reaction conditions of the embodiment as shown in Table 3;

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

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Abstract

The present invention relates to a bovine milk β-lactoglobulin T cell immune tolerance hydrolyzate and a preparation method and application thereof. The method includes predicting β-lactoglobulin T cell epitopes; using different kinds of proteases to enzymatically hydrolyze β-lactoglobulin, and using mass spectrometry to determine the amino acid sequence of the hydrolyzed peptide fragments, and comparing the amino acid sequences of these peptide fragments with the predicted amino acid sequences The β-lactoglobulin T cell epitope sequences were compared, and the hydrolysates that may have immune tolerance were initially screened; the screened hydrolysates that may have immune tolerance were used as the research objects, and animal experiments were used to further screen and A hydrolyzate that does have immune tolerance. The present invention successfully establishes a model of oral tolerance induced by trypsin, composite protease and papain beta-lactoglobulin hydrolyzate in Balb / c mice, and these three hydrolyzates can effectively prevent OVA-induced allergic reaction in mice. According to the β-lactoglobulin hydrolyzate of the present invention, peptide drugs or functional foods can be developed.

Description

technical field [0001] The present invention relates to the technical field of food biotechnology, in particular to a bovine milk beta-lactoglobulin T cell immune tolerance hydrolysate and a preparation method and application thereof. Background technique [0002] With the continuous development of the food industry, the safety of food has been paid more and more attention by people. In recent years, the phenomenon of food allergy has increased year by year, and a large number of studies have shown that among food allergies, cow's milk protein allergy is the most common. Cow's 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 nutrients for the body. However, milk allergy seriously affects the food safety of people with allergies. Nearly 20% of the total protein in milk is whey protein, and alpha-lactalbumin and beta-lactoglobulin are t...

Claims

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Application Information

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Patent Type & Authority Patents(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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