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Method for separating beta-lactoglobulin from raw milk

The technology of lactoglobulin and raw milk is applied in the field of separation and purification of beta-lactoglobulin, which can solve the problems of complicated instrument, small processing capacity and high preparation cost, and achieve the effect of improving separation efficiency

Inactive Publication Date: 2012-10-24
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are methods for separating β-lactoglobulin, such as affinity chromatography, preparative chromatography, dialysis, etc., but the processing volume is small, the equipment is complicated, and the preparation cost is high

Method used

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  • Method for separating beta-lactoglobulin from raw milk
  • Method for separating beta-lactoglobulin from raw milk

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Embodiment 1

[0033] Embodiment 1. A method for isolating β-lactoglobulin from raw milk, using 1000ml of fresh cow's milk as a raw material; following steps in sequence:

[0034] 1) Fresh milk is centrifuged at 4000g at 4°C for 30 minutes, filtered through 4 layers of skimmed gauze to remove milk fat, and the first filtrate obtained is collected;

[0035] 2) Adjust the pH of the first filtrate to 4.6 with 1 M hydrochloric acid, and let it stand at room temperature for 30 minutes; obtain the first filtrate after treatment;

[0036] 3) Centrifuge the filtrate for the first time after treatment at 4°C, 5000g for 20 min, and filter it with 4 layers of degreasing gauze. gauze filter), to obtain the secondary filtrate;

[0037] 4) Adjust the pH of the secondary filtrate to 2.0 with 1M hydrochloric acid to obtain the treated secondary filtrate; record the volume of the treated secondary filtrate;

[0038] 5) First add 70g of sodium chloride to the secondary filtrate according to the dosage ratio...

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Abstract

The invention discloses a method for separating beta-lactoglobulin from raw milk. The method comprises the following steps: with fresh cow's milk as a processing object, carrying out centrifugation and degreasing so as to obtain skim milk; adding an acid to adjust the pH value of the skim milk, standing the skim milk for a certain period of time and then removing casein through centrifugation; adjusting the pH value of a solution so as to allow the solution to be acidic, adding a salt, standing the solution for a certain period of time, then carrying out centrifugation and collecting supernatant; adjusting the pH value of the supernatant, adding a sodium salt, carrying out centrifugation after standing and collecting precipitate; flushing the precipitate and then dissolving the precipitate in a PBS buffer solution; carrying out ultrafiltration on the solution to remove salt and carrying out freeze-drying; and identifying beta-lactoglobulin by using sodium dodecanesulphonate-polyacrylamide gel electrophoresis and quantifying purity by using reverse-phase chromatography. According to the method provided in the invention, beta-lactoglobulin can be separated from fresh cow's milk through acid deposition and salt precipitation, and desalting through ultrafiltration is combinedly used so as to obtain high purity beta-lactoglobulin. The method improves separation efficiency of beta-lactoglobulin and provides a high-quality raw material for research on other aspects of and application of beta-lactoglobulin.

Description

technical field [0001] The invention relates to a method for separating and purifying beta-lactoglobulin, in particular to a method for efficiently separating and purifying beta-lactoglobulin from fresh milk. Background technique [0002] β-lactoglobulin exists in the milk of most mammals. It is a milk-specific protein synthesized by mammary epithelial cells. The concentration in bovine milk is about 3.2g / L, mainly in the form of dimers, and 2 monomers linked by non-covalent bonds. At pH<3.0 or pH>7.0, β-lactoglobulin dimers dissociate into monomers. The molecular weight of monomeric β-lactoglobulin is about 18300 Daltons, and the isoelectric point is 5.1-5.3. β-lactoglobulin accounts for about 55% of whey protein, is the main protein in whey, and is the functional embodiment of whey protein. It has biological activities such as changing the thermal stability of milk, combining with fatty acids and flavor substances, anti-cancer, etc., and has good processing proper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/36C07K1/34C07K1/30C07K1/20
Inventor 刘建新任大喜缪淑颖
Owner ZHEJIANG UNIV
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