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Preparation method of high-purity casein phosphopeptide

A casein phosphopeptide, high-purity technology, applied in the field of preparation of high-purity casein phosphopeptide, can solve the problems of low yield and content of casein phosphopeptide, low reaction temperature, low equipment requirements, etc., and achieve low cost, Good water solubility and simple process

Active Publication Date: 2014-02-12
GREENCREAM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method is simple in operation, low in equipment requirements, high in yield, good in product quality, and low in reaction temperature, and can realize industrialized production, but it adopts a single protease enzymolysis, and the yield and content of casein phosphopeptides are low (the highest product content is about 16%), the product has not been debittered

Method used

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  • Preparation method of high-purity casein phosphopeptide
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  • Preparation method of high-purity casein phosphopeptide

Examples

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Effect test

Embodiment 1

[0021] Example 1 : a kind of preparation of high-purity casein phosphopeptide, comprises the following steps:

[0022] Add 150L of pure water to the reaction kettle, start stirring at 200rpm, heat to 50°C, slowly put 25Kg of casein into the reaction kettle, and adjust the pH to 7.8; put in trypsin: neutral protease: alkaline protease: chymotrypsin = 12.5g of compound protease made by mixing 4:3:2:1 (mass ratio). After 1 hour of enzymatic hydrolysis, the temperature of the feed solution was lowered to 37°C, the pH was adjusted to 4.0, 12.5 g of pepsin was added, and the enzymatic hydrolysis was continued, and the enzymatic hydrolysis ended after 1 hour. Adjust the pH to 4.5 to inactivate the enzyme, adjust the temperature to 90°C, and incubate for 20 min. Use 732 cation exchange resin to absorb and purify for 1 h, filter, adjust the pH of the purified solution to 7.0 with the filtrate, use 717 anion exchange resin to purify for 1 h, wash with water first, discard the washing...

Embodiment 2

[0023] Example 2 : a kind of preparation of high-purity casein phosphopeptide, comprises the following steps:

[0024] Add 200L of pure water to the reaction kettle, stir at 200rpm, heat to 45°C, slowly put 25Kg of casein into the reaction kettle, adjust the pH to 7.5; put in trypsin: neutral protease: papain: chymotrypsin = 3 : 25g of complex protease made by mixing 2:4:1. After 1.5 hours of enzymatic hydrolysis, the temperature of the feed liquid was lowered to 40°C, the pH was adjusted to 5, 25g of pepsin was added, and the enzymatic hydrolysis was continued, and the enzymatic hydrolysis was completed after 1 hour. Adjust the pH to 4.6, adjust the temperature to 80°C, and incubate for 30 min to inactivate the enzyme. Filter to remove unenzymolyzed casein and other macromolecular protein precipitates, use 001×7 cation exchange resin to absorb and purify the supernatant for 1 hour, filter, adjust the pH of the purified solution to 6.5 with the filtrate, and use American Am...

Embodiment 3

[0025] Example 3 : a kind of preparation of high-purity casein phosphopeptide, comprises the following steps:

[0026] Add 250L of pure water to the reaction kettle, stir at 200rpm, heat to 45°C, slowly put 25Kg of casein into the reaction kettle, and adjust the pH to 8.5; put in trypsin:alkaline protease:papain:neutral protease=2 : 50g of compound protease made by mixing 3:1:4. After 2 hours of enzymatic hydrolysis, the temperature of the feed solution was lowered to 37°C, the pH was adjusted to 4.5, 12.5 g of pepsin was added, and the enzymatic hydrolysis was continued, and the enzymatic hydrolysis ended after 1 hour. Adjust the pH to 4.7, adjust the temperature to 85°C, incubate for 30 min, and inactivate the enzyme. Filter to remove unenzymolyzed casein and other macromolecular protein precipitates, use Rohm and Haas AMBERJET UP6040 cation exchange resin to absorb and purify the supernatant for 1 hour, filter, adjust the pH of the purified solution to 7.0 with the filtr...

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Abstract

The invention discloses a preparation method of high-purity casein phosphopeptide. The method comprises the following steps: adding water to dissolve casein; carrying out secondary enzymolysis by using compound protease and pepsase prepared from any four of trypsase, neutral protease, alkaline protease, curd protease and nematolyt; carrying out secondary purification treatment through cation exchange resin and anion exchange resin after sterilizing and filtering; washing hydrolysate by water; eluting by hydrochloric acid of which the concentration is 0.5-1%; desalting and drying by a nanofiltration membrane. By adopting the preparation method, the yield (greater than or equal to 17%) and the purity (greater than or equal to 90%) of the casein phosphopeptide can be effectively improved; the prepared casein phosphopeptide is bitter in taste, and good in water-solubility; a water solution containing 20% of casting polypropylene (CPP) is clear and transparent; the casein phosphopeptide is powder of which the color is pure white. The molecular weight of the milk casein phosphopeptide prepared by the method mainly is between 1000 and 5000; absorption of a human body is facilitated; the preparation method disclosed by the invention is a preparation method which is low in cost, simple in process, and applicable to industrial production.

Description

technical field [0001] The invention relates to a method for preparing high-purity casein phosphopeptides using casein in cow milk as raw material. Background technique [0002] Casein phosphopeptide (CPP) is made from cow's milk casein through biotechnology. It has peptides that promote the absorption and utilization of divalent metal ions such as calcium, iron, and zinc. It can be used in various nutritional and health foods. Casein phosphopeptide is made by hydrolyzing casein with trypsin or trypsin, refined and purified, and its core structure is: -Ser(P)-Ser(P)-Ser(P)-Glu- G lu - (Ser: serine, Glu: glutamic acid, P: phosphate group). The phosphoserine residues (-Ser(P)-) in this structure exist in clusters, which are negatively charged in the weak alkaline environment of intestinal pH, which can prevent the further action of digestive enzymes, so that casein phosphopeptides will not was further hydrolyzed. Casein phosphopeptides can combine with calcium in the enviro...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/18
Inventor 曹庸李双祁刘飞彭维郭斌吴成顺黄慧明王海波
Owner GREENCREAM BIOTECH
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