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Immobilization method of beta-galactosidase

A technology of galactosidase and enzyme solution, which is applied in the field of immobilization of β-galactosidase, can solve problems such as the reduction of immobilization efficiency, and achieve the effects of simplifying operations, increasing reuse rates, and reducing costs

Active Publication Date: 2021-03-09
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, Eskandarloo et al. [Eskandarloo Hamed et al., Production of galacto-oligosaccharides from whey permeate using β-galactosidase immobilized on functionalized glass beads [J]. Food Chemistry 251 (2018) 115-124.] used glass beads with a diameter of 5 mm as the enzyme In the immobilization process of the low-concentration enzyme solution (2mg / ml), the immobilization efficiency of 80% was achieved, but the immobilization efficiency decreased with the increase of the enzyme concentration

Method used

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  • Immobilization method of beta-galactosidase
  • Immobilization method of beta-galactosidase
  • Immobilization method of beta-galactosidase

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Embodiment 1, a kind of immobilization method of β-galactosidase, comprises the following steps:

[0054] 1), preparation of polyethyleneimine solution

[0055] Dilute polyethyleneimine mother liquor (500mg / ml) with deionized water, obtain the polyethyleneimine dilution of 0.25mg / ml, then utilize the sodium hydroxide of 3mol / L or hydrochloric acid to adjust the pH=8 of this dilution, as Polyethyleneimine solution, store at room temperature.

[0056] 2) Adsorption and cross-linking of β-galactosidase

[0057] Take 1ml of the above-mentioned polyethyleneimine solution (at room temperature) and put it in a 5ml centrifuge tube, soak a 0.1g rectangular cotton face towel (referred to as the face towel) in it and let it stand for 5 minutes; take out the face towel, if the polyethyleneimine solution remains in the centrifuge tube , then use a pipette to absorb it and add it all dropwise to the face towel; transfer the face towel to a new centrifuge tube, and pipette to absorb...

Embodiment 2

[0071] Embodiment 2, a method for producing galactooligosaccharides by immobilization of β-galactosidase, which expands the immobilized enzyme production system on the basis of Example 1, and increases the number and time of continuous testing of the immobilized enzyme , to further provide favorable evidence for the operational stability of the immobilized enzyme under this method; it mainly includes the following steps:

[0072] 1), preparation of polyethyleneimine solution

[0073] With embodiment 1.

[0074] 2) Adsorption and cross-linking of β-galactosidase

[0075] Take 2.5ml of the above-mentioned polyethyleneimine solution (at room temperature) and put it in a 10ml centrifuge tube, soak a 0.4g rectangular cotton face towel in it and let it stand for 10min; if the polyethyleneimine solution remains in the centrifuge tube, use a pipette After aspirating, add it dropwise to the face towel; transfer the face towel to a new centrifuge tube, pipette 2ml of 10mg / ml pre-coole...

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Abstract

The invention discloses an immobilization method of beta-galactosidase. The method comprises the following steps: regulating a pH value of a polyethyleneimine diluent with a concentration of (0.25 + / -0.05) mg / ml to (8 + / -0.2), taking 0.1-0.4 g of cotton cloth, adding 1-2.5 ml of a polyethyleneimine solution, and performing standing for 5-10 min; adding 1-2 ml of a pre-cooled beta-galactosidase enzyme solution with at a concentration of 5-10 mg / ml and 3-4 ml of deionized water, and performing shaking for (10 + / -1)min by a shaker; adding 1-2.5 ml of a pre-cooled 0.2% (v / v) glutaraldehyde aqueoussolution, and performing shaking for (5 + / -1) min by a shaker; washing an obtained product so as to wash off a free enzyme which is not combined to the cotton cloth; and obtaining the cotton cloth loaded with the galactosidase.

Description

technical field [0001] The invention relates to a method for immobilizing β-galactosidase, which belongs to the technical field of immobilized enzymes. Background technique [0002] Galactooligosaccharides have aroused great interest among researchers due to their extensive prebiotic effects. Although galactooligosaccharides are produced in a variety of ways, they are commonly obtained by enzymatic hydrolysis with β-galactosidase. . The full name of β-galactosidase is β-D-galactoside galactohydrolase, referred to as lactase. β-galactosidase exists in various animals, plants and microorganisms, and is widely used in the fields of food, immunity and environmental detection. [0003] At present, some manufacturers at home and abroad use free lactase to produce galacto-oligosaccharides, which is produced by one-time addition of enzymes, which makes it difficult to recycle and reuse enzymes, resulting in increased costs. For these problems, immobilized lactase highlights its u...

Claims

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Application Information

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IPC IPC(8): C12N11/02
CPCC12N11/02C12N9/2471C12Y302/01023
Inventor 王敏奇王庚
Owner ZHEJIANG UNIV
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