A method for extracting lactoperoxidase using ultrafiltration-assisted two-phase extraction technology

A technology of lactoperoxidase and extraction technology, applied in the direction of oxidoreductase, biochemical equipment and methods, enzymes, etc., can solve the problem of lack of lactoperoxidase

Active Publication Date: 2017-11-28
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Domestic studies have reported the extraction of lipase, glucoamylase, amylase and other processes using the two-phase method, but there is no report on the application of the two-phase method to extract lactoperoxidase

Method used

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  • A method for extracting lactoperoxidase using ultrafiltration-assisted two-phase extraction technology
  • A method for extracting lactoperoxidase using ultrafiltration-assisted two-phase extraction technology
  • A method for extracting lactoperoxidase using ultrafiltration-assisted two-phase extraction technology

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Experimental program
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Embodiment 1

[0039] The extraction of embodiment 1 lactoperoxidase

[0040] (1) Aqueous two-phase extraction lactoperoxidase:

[0041] The whey is obtained after removing the fat and casein in the fresh milk. At room temperature, add an appropriate amount of whey to the container, directly add PEG and salt to the whey continuously and place it on a stirrer for continuous stirring. Until the PEG and salt are completely dissolved in the whey, an aqueous two-phase system is formed. Among them, the molecular weight of PEG is 6000Da, and the salt is MgSO 4 , the final concentration of PEG is 16% (w / w), MgSO 4 The final concentration of the solution was 10% (w / w), and the pH of the system was adjusted to 8.5 with 1 mol / L NaOH, and stood still for about 30 minutes. After the two phases were completely separated, the lower phase of the system, namely the salt phase, was collected.

[0042] (2) Ultrafiltration: Use 1mol / L HCl to adjust the pH of the collected salt phase solution system to about ...

Embodiment 2

[0059] The extraction of embodiment 2 lactoperoxidase

[0060] (1) Aqueous two-phase extraction lactoperoxidase:

[0061] The whey is obtained after removing the fat and casein in the fresh milk. At room temperature, add an appropriate amount of whey to the container, directly add PEG and salt to the whey and place it on a magnetic heating stirrer continuously Stir until the PEG and salt are completely dissolved in the whey to form an aqueous two-phase system. The molecular weight of PEG is 6000Da, and the salt is MgSO 4 , the final concentration of PEG is 14% (w / w), MgSO 4 The final concentration of the solution is 12% (w / w). Use 1 mol / L NaOH to adjust the pH of the system to 8.0. Stand still in the separatory funnel for about 30 minutes until the two phases are completely separated, and collect the lower phase of the system, that is, the salt phase.

[0062] (2) Ultrafiltration: Use 1mol / L HCl to adjust the pH of the salt phase solution system of the collected lactoperoxi...

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Abstract

The invention discloses a method for extracting lactoperoxidase by the ultrafiltration assisted two aqueous phase extraction technology. The method comprises the steps of using bovine whey as the raw material; enriching all lactoperoxidase from bovine whey into a salt phase by the two aqueous phase extraction technology; processing the salt phase solution through an ultrafiltration membrane to remove small impure protein; further purifying lactoperoxidase; desalting through a dialysis bag; then freezing and drying, so as to prepare a high-activity lactoperoxidase product which can be used in natural food additives. With the adoption of the method, lactoperoxidase with natural antibacterial activity can be directly prepared and directly added as a preservative into foods or toothpaste and other daily supplies.

Description

technical field [0001] The invention relates to a method for extracting lactoperoxidase, in particular to a method for extracting lactoperoxidase by using an ultrafiltration-assisted two-phase extraction technique. The invention belongs to the technical field of extraction and preparation of lactoperoxidase. Background technique [0002] Lactoperoxidase (LP) is a natural antibacterial substance present in animal milk. In the presence of hydrogen peroxide, lactoperoxidase can catalyze thiocyanate to form an antibacterial medium. The biology of LP The meaning is to participate in the natural host's defense against the invasion of microorganisms, and to protect the mammary glands of animals during lactation and the intestinal pathogenic microorganisms of newborn babies. LP is composed of a single polypeptide chain containing 612 amino acids, with a molecular weight of about 85kDa, etc. The electric point is 8.0-8.5, and the content in milk is 10-30mg / mL. It has been proved in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08
CPCC12N9/0065C12Y111/01007
Inventor 姜瞻梅许晓曦刘丽丽车红霞付红岩
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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