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Preparation method of milk-source calcium phytochelatin

A technology of chelating peptide and milk-derived calcium, which is applied in the field of functional polypeptide preparation, can solve the problems of high gastrointestinal irritation, low biological titer, low calcium content, etc., and achieves the promotion of absorption, improvement of calcium absorption, and good physiological activity. Effect

Inactive Publication Date: 2020-01-07
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Inorganic calcium salts, as the first generation of calcium supplement products, have low biological potency and are highly irritating to the stomach; organic calcium salts, as the second generation of calcium supplement products such as calcium gluconate and calcium lactate, are less irritating to the stomach but low in calcium content , you have to take dozens of tablets every day to meet the calcium supplement requirements; amino acid chelated calcium is easy to be absorbed by the human body, and successfully overcomes the shortcomings of inorganic calcium salts and organic acid calcium, but the production cost is high and it is difficult to popularize

Method used

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  • Preparation method of milk-source calcium phytochelatin
  • Preparation method of milk-source calcium phytochelatin
  • Preparation method of milk-source calcium phytochelatin

Examples

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specific Embodiment 1

[0025] Specific embodiment 1: the present invention provides a kind of method for preparing milk-derived calcium chelating peptide, and this calcium chelating peptide can be prepared by following preparation method:

[0026] (1) Enzymolysis of casein: Weigh 100g of casein, place it in a 1000mL beaker, add tertiary water and 7 mL sodium hydroxide solution (1mol / L, to speed up the dissolution), and put it in 50 Stir and dissolve in a constant temperature water bath at ℃ for 30 min, 120r / min. After taking it out, use sodium hydroxide (1mol / L) solution to accurately adjust the pH to 8.0, then add 0.1g of trypsin, and perform enzymolysis in a water bath for 3 hours (50°C, 120r / min). After enzymolysis, take it out and place it in a boiling water bath to inactivate the enzyme for 10 minutes, then place it at room temperature to cool, then adjust the pH to 4.6 with hydrochloric acid (1mol / L) solution after cooling, and use the isoelectric point to precipitate unenzymolyzed casein. Th...

Embodiment 2

[0043] Example 2: In this example, the effect of calcium peptide chelate on calcium transport in Caco-2 cell monolayer was studied, the specific process is as follows.

[0044] (1) Cell culture

[0045] Caco-2 cells were purchased from American Type Culture Collection (Rockville, MD, USA). Caco-2 cells of passage 40 to 44 were used in the experiment. Cells were cultured in complete medium (10% fetal calf serum, 1% non-essential amino acid, 1% double antibody added to EMEM).

[0046] When the cells grew to 90% confluency, Caco-2 cells were separated by trypsin-EDTA treatment, and plated on a 6-well transwell culture plate (3450, Corning Inc, USA) with a polyester membrane (24 mm in diameter) at 3× 10 5 Inoculate at a density of cells / mL. The apical and basolateral media were changed every other day for 21 days. The integrity of the Caco-2 cell monolayer was confirmed by measuring the transepithelial electrical resistance (TEER) using an electrical resistance device (Millic...

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Abstract

The invention provides a preparation method of milk-source calcium phytochelatin. Specifically, cow milk casein is selected as a raw material, trypsin is adopted as a tool enzyme, a calcium-peptide chelate is prepared by adding a calcium salt into zymolyte, then the peptide-calcium chelate is decalcified through EDTA disodium, and thus polypeptide with the calcium chelating activity is obtained. The preparation method is efficient, operation is easy, very high social benefits are achieved, and the prepared milk-source protein calcium phytochelatin has the high calcium chelating activity, is stable in property, is a food-source peptide, is natural and safe and is an ideal functional health-care product additive.

Description

technical field [0001] The invention relates to a preparation method of calcium chelating peptide, in particular to the separation and purification of calcium chelating peptide from bovine milk casein hydrolyzate, and belongs to the field of functional polypeptide preparation. Background technique [0002] Milk is rich in protein, 80% of which is casein. The bovine milk casein undergoes enzymatic hydrolysis and separation and purification to obtain active peptides: casein phosphopeptides, referred to as CPPs. CPPs are a class of polypeptides rich in phosphoserine and glutamic acid, so CPPs have a strong ability to chelate calcium ions. A large number of studies have shown that CPPs have a variety of physiological functions, and are known as "mineral carriers" because of their good function of promoting mineral absorption. It has been proved by experiments that CPPs can significantly accelerate the metabolism of calcium and increase the storage of calcium, and the calcium a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/107
CPCC07K1/107C12P21/06
Inventor 康梦谢凤英苗建银
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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