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Method for fractional precipitation and purification of casein

A technology of fractional precipitation and casein, applied in the field of casein separation and purification, can solve the problems of low production efficiency, complex process, unsuitable for large-scale industrial preparation, etc., and achieves high recovery rate, good application prospect, simple and easy operation Effect

Active Publication Date: 2021-10-01
湖北百特威生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The casein separation method in the prior art is complex in process and low in production efficiency, such as the chromatography in CN107840883, which is mainly used in the field of scientific research and is not suitable for large-scale industrial preparation, and toxic reducing agents will be added to the buffer used for separation. agent, generally not used for the separation of food-grade raw materials

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Take a certain mass of crude casein, add a certain volume of ethanol solution with a mass concentration of 50%, fully dissolve it and centrifuge for 20 minutes to take the supernatant, and obtain the k-casein component after the supernatant is rotary steamed; then take the obtained precipitate, precipitate It is a mixture of α-casein and β-casein, dissolve in water at 20°C, adjust the pH to neutral, cool at 0°C for 2 hours, adjust the pH to 3.5, centrifuge at 8000rpm for 20min, the precipitate after centrifugation is α - casein fraction; the supernatant was raised to room temperature, adjusted to pH 3.5, centrifuged at 8000 rpm for 20 min, the precipitate was collected, and the β-casein fraction was obtained after washing. Finally, 39% of alpha-casein fraction, 23% of beta-casein fraction and 10% of k-casein fraction are obtained, with a purity of over 95%.

Embodiment 2

[0017] Take a certain mass of crude casein, add a certain volume of ethanol solution with a mass concentration of 60%, fully dissolve it, and centrifuge it for 20 minutes to take the supernatant, and obtain the k-casein component after the supernatant is rotary steamed; It is a mixture of α-casein and β-casein, dissolve in water at 20°C, adjust the pH to neutral, cool at 2°C for 4 hours, adjust the pH to 3.5, centrifuge at 8000rpm for 20min, the precipitate after centrifugation is α - casein fraction; the supernatant was raised to room temperature, adjusted to pH 3.5, centrifuged at 8000 rpm for 20 min, the precipitate was collected, and the β-casein fraction was obtained after washing. Finally, 43% of alpha-casein fraction, 25% of beta-casein fraction and 8% of k-casein fraction are obtained, with a purity of over 95%.

Embodiment 3

[0019] Take a certain mass of crude casein, add a certain volume of ethanol solution with a mass concentration of 60%, fully dissolve it, and centrifuge it for 20 minutes to take the supernatant, and obtain the k-casein component after the supernatant is rotary steamed; It is a mixture of α-casein and β-casein, dissolve in water at 20°C, adjust the pH to neutral, cool at 2°C for 4 hours, adjust the pH to 4.5, centrifuge at 8000rpm for 20min, the precipitate after centrifugation is α - casein fraction; the supernatant was raised to room temperature, adjusted to pH 4.5, centrifuged at 8000 rpm for 20 min, and the precipitate was collected, washed to obtain the β-casein fraction. Finally, 47% of α-casein component, 30% of β-casein component and 11% of k-casein component are obtained, with a purity of over 95%.

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Abstract

The invention belongs to the field of separation and extraction of casein. The method comprises the steps: separating alpha-, beta-and k-casein by a simple and feasible ethanol separation, isoelectric precipitation and low-temperature cooling method; weighing a certain mass of casein, adding a certain volume of an ethanol solution, fully dissolving, centrifuging for 20 minutes, taking supernate, and carrying out rotary evaporation on the supernate to obtain a k-casein component; dissolving the obtained precipitate in water at the temperature of 20 DEG C, adjusting the pH value to 7.0, cooling for 2-6 hours at the temperature of 0-4 DEG C, adjusting the pH value to 3.5-5.5, centrifuging for 20 minutes to obtain alpha-casein precipitate, and repeatedly dissolving and precipitating to obtain high-purity alpha-casein; heating the supernatant to room temperature, adjusting the pH value to 3.5-5.5, and performing centrifuging to obtain the beta-casein component.

Description

technical field [0001] The invention belongs to the field of separation and purification of casein, in particular to the separation and extraction of alpha-casein, beta-casein and k-casein in casein. Background technique [0002] Casein is a kind of phosphorus-calcium-binding protein in mammalian milk. It is rich in content, equivalent to 2.75% of the total milk composition, and contains 8 kinds of amino acids necessary for human body, with an isoelectric point of about 4.6, including α s1 -, α s2 -, β-, k-casein four main monomers, α-casein accounts for about 50% of the total casein, β-casein accounts for about 34% of the total casein. Casein does not exist in the form of a single component in milk, but cross-links with each other to form spherical aggregates, that is, casein micelles. There are significant differences in the structure and properties of each component of casein, and a single casein The components can better reflect the functions of casein in some function...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K1/30C07K1/14
CPCC07K14/4732
Inventor 王冠华许涌捷伍昌军
Owner 湖北百特威生物科技有限公司
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