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Method for separating milk-derived whey proteins in PEG/phosphate aqueous two-phase system

An aqueous two-phase system and whey protein technology, which is applied in the field of separating milk-derived whey protein α-La and β-Lg by a PEG/phosphate aqueous two-phase system, and achieves the advantages of protein biological activity, low price, and easy industrialization. production effect

Active Publication Date: 2014-01-22
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented process describes an improved way to isolate pure cow's milk with specific ingredients such as beta lactoglobulin or lysolecithin. It uses this purified product instead of traditional methods like precipitation techniques or ion exchange resin columns. These new processes have several benefits including being costly and efficient at producing high quality baby formulas containing these substances.

Problems solved by technology

The technical problem addressed in this patented text relates to improving the quality of wheyside dairy product containing both beta -galactosylceramia (BGI) and alpha lipid glycocyanobilitrite (ALC). Beta galanoproteina (BGA) is one type of vitamin present naturally occurring within animal tissues such as brain cells, muscles, teeth, nails, skin, lymbacillis, heart valves, skeletons, joints, blood vessels, fats, urolithochlorides, steroids, glucans, peptones, phosphorus compounds, and oligopea components. Additionally, there may contain some harmful metabolites like trypsin disulfatase, lysozodium salt, yarnule albumen, sialloprotein, serotoxins, iron defeclux pills, mangellose dietary fiber, magnesiophyllite, and polygalaxilage. These contributes to poor taste sensory attributes associated with these materials in infants who consume them regularly throughout childhood life.

Method used

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  • Method for separating milk-derived whey proteins in PEG/phosphate aqueous two-phase system
  • Method for separating milk-derived whey proteins in PEG/phosphate aqueous two-phase system
  • Method for separating milk-derived whey proteins in PEG/phosphate aqueous two-phase system

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Experimental program
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Effect test

Embodiment 1

[0039] Whey protein was separated by PEG / phosphate two-phase system. The separation steps are as follows:

[0040] (1) Preparation of standard whey protein solution: standard α-La solution (1mg / mL); standard β-Lg solution (1mg / mL).

[0041] (2) Southern buffalo milk whey protein extraction: Commercially available buffalo milk was centrifuged at 4000r / min at 4°C for 20 minutes to skim, and the upper layer of fat was discarded. The skim milk was adjusted to pH 4.6 with 5M HCl, and stood for 30min at 4000r / min at 4°C. Centrifuge for 20 minutes, discard the precipitate, filter the supernatant with a 0.22 μm filter membrane, dialyze against distilled water at 4°C overnight, freeze-dry, and store in a freezer at -20°C.

[0042] (3) Use the cloud point method to draw the phase diagram of the two-phase aqueous system. The experimental method is as follows: Take a certain mass of inorganic salt (m1) and add a certain amount of distilled water (m2), dissolve, and constant volume, and c...

Embodiment 2

[0053] Embodiment 2: analyze the phase forming ability of four kinds of PEG and phosphate

[0054] In the step (3) of Example 1, the cloud point method was used to make the two-phase phase diagram of PEG1000, 1500, 4000, 6000 and dipotassium hydrogen phosphate to understand the phase-forming ability of the four PEGs and phosphate. The longer the two-node line in the aqueous two phase diagram, the stronger the phase separation ability of the two substances. The phase diagram of PEG and phosphate is as follows figure 2 shown.

Embodiment 3

[0055] Embodiment 3: Determination of PEG mass fraction in the best PEG type and system

[0056] Prepare the PEG / phosphate two-phase system with PEG mass fractions of 10.4%, 11.4%, 12.4%, 13.4%, and 14.4%, PEG types of 1000, 1500, and 4000, phosphate concentration of 14.8%, and system pH of 7.3 For the extraction effect of α-La and β-Lg, calculate the distribution coefficient and recovery rate of α-La and β-Lg respectively. The larger the ratio of the distribution coefficient of α-La and β-Lg, the better the separation effect. Best PEG species and PEG quality scores. Among them, the effect of PEG type and mass fraction on the distribution coefficient of β-Lg, and the effect of PEG type and mass fraction on the distribution coefficient of α-La are shown in Fig. image 3 and Figure 4 shown.

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Abstract

The invention discloses a method for separating milk-derived whey proteins of alpha-La and beta-Lg in a PEG/phosphate aqueous two-phase system, and belongs to the whey protein components separation technology. The milk-derived whey proteins are extracted; alpha-La and beta-Lg are separated in the PEG/phosphate aqueous two-phase system, wherein in the aqueous two-phase system, the mass fraction of PEG is 10.4%-14.4%, the concentration of phosphate is 10.8%-18.8%, and the pH of the system is 7.3; then, a coomassie brilliant blue method is used for measurement of the protein concentration, and SDS-PAGE electrophoresis is used for identification and detection. The method disclosed by the invention provides the whey protein separation technology which is cheap, simple in operation and easy to amplify and can be applied to large-scale industrialized production; and the milk-derived whey proteins of alpha-La and beta-Lg obtained by separation are applied in production of functional food and bioactivity products.

Description

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Claims

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Application Information

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Owner JINAN UNIVERSITY
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