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Small molecular milk casein polypeptide and preparing method

A technology of casein and small molecules, which is applied in the field of casein peptides, small molecule casein peptides and their preparation, which can solve the problems of affecting the taste, losing the nutrition and efficacy of the original protein, etc.

Inactive Publication Date: 2015-11-25
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, small molecular proteins tend to lose the nutrition and efficacy of the original protein
Moreover, the amino acid itself has a bitter taste, the more fully decomposed and the higher the purity of the amino acid, the more bitter the taste, therefore, it will affect the taste

Method used

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  • Small molecular milk casein polypeptide and preparing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Mix 1L of milk with 50ml of papain, centrifuge at 2000rpm for 5 minutes to obtain a coagulated precipitate; add the coagulated precipitate to water, boil, centrifuge at 2000rpm for 5 minutes, and take the supernatant; add the supernatant to an acidic buffer of pH 4-5 solution, stirred, and centrifuged at 10,000rpm for 30 minutes to obtain a precipitate; the precipitate was dissolved in a pH4.5-5.0 buffer, and the SephadexG25, pH7.2 PBS buffer was used as the mobile phase to separate a molecular weight of 2-3KDa The polypeptide is obtained. The sequence of the polypeptide determined by a solid-phase protein automatic sequencer is VIMYQILPQNFLPILQPAVBVPFLQ, and the purity is 82%.

Embodiment 2

[0013] Mix 1L of milk with 100ml Albizia juniper protease, centrifuge at 3000rpm for 10 minutes to get coagulated precipitate; add the coagulated precipitate to water, boil, centrifuge at 3000rpm for 10 minutes, take the supernatant; add the supernatant to the acidic buffer of pH 4-5 solution, stirred, and centrifuged at 15000rpm for 60 minutes to obtain a precipitate; the precipitate was dissolved in a pH4.5-5.0 buffer, and the SephadexG25, pH7.2 PBS buffer was used as the mobile phase to separate the molecular weight of 2-3KDa The polypeptide is obtained. The sequence of the polypeptide determined by a solid-phase protein automatic sequencer is VIMYQILPQNFLPILQPAVBVPFLQ, and the purity is 78%.

Embodiment 3

[0015] Immunogenicity test: 6 BALB / c white mice, half male and half male, were orally administered with 36 mg / kg of the polypeptide prepared in Example 1, respectively. The drug was administered continuously for 8 weeks, and blood was collected from the orbital venous plexus once a week from 1 to 12 weeks, centrifuged at 12,000 rpm for 2 minutes, the plasma was separated to obtain the supernatant, and stored at -20°C for later use. After dissolving at room temperature, take 0.1 ml of the supernatant to detect the antibody titer in the serum by indirect ELISA, wherein the ELISA detection kit was purchased from Shanghai Sangong. The RA value is obtained by comparing the actual A value (OD450) of each sample with the serum A value of the negative control. RA ≥ 2.1 is positive. Bloodbio-samples are tested after a series of dilutions, and the highest dilution with a positive reaction is the The titer of the sample. The measurement result is displayed. The blood samples collected ...

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PUM

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Abstract

The invention discloses a small molecular milk casein polypeptide, mainly relating to a small molecular casein polypeptide and a preparation method. The sequence of the small molecular milk casein polypeptide is VIMYQILPQNFLPILQPAVBVPFLQ. Preparing steps comprise: (1) mixing 1L of milk with 50-100ml of chymosin and centrifuging at 2,000-3,000rpm for 5-10min to obtain solidification precipitate; (2) adding the solidification precipitate in water, boiling, and centrifuging at 2000-3000rpm for 5-10min to take supernatant; (3) adding supernatant into an acid buffer solution of pH being 4-5, stirring, centrifuging at 10,000-15,000rpm for 30-60min to obtain precipitate; (4) dissolving the precipitate into a buffer solution of pH being 4.5-5.0, separating polypeptide with molecular weight being 2-3KDa through Sephadex G25. The method has the beneficial effects that the prepared polypeptide has purity more than 60%, and can obviously promote absorption of trace elements of Ca and Mg, which is equivalent to the casein, and effects of no antigenicity and good mouth feeling are realized.

Description

technical field [0001] The invention relates to the field of casein polypeptides, in particular to the field of small molecule casein polypeptides and their preparation. Background technique [0002] Milk is a high-quality nutritious food, but it is also one of the foods that are more likely to cause allergies. Casein accounts for 80% of the total milk protein, is the protein with the highest content in milk, and is also the most allergenic protein among the five allergenic proteins in milk. Heterosexual proteins, such as casein, because their molecular weight is too large, after entering the body, the gastrointestinal tract of immature babies lacks the enzymes to decompose them, and they enter the blood circulation as antigens, stimulating the body to produce antibodies, thus causing allergic reactions, such as abdominal pain, Symptoms such as abdominal distension or diarrhea, asthma, urticaria, eczema, allergic rhinitis, mucus and blood in stool. These symptoms can cause...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12P21/06C07K1/14
CPCC07K14/4732
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH