Co-crosslinking immobilization method of beta-galactosidase

A galactosidase and co-crosslinking technology, applied in the directions of immobilized enzymes, biochemical equipment and methods, glycosylases, etc., can solve the problems of high reaction activity, excessive crosslinking of enzymes, and waste of enzymes, and improve the Microenvironment and the effect of improving catalytic activity

Active Publication Date: 2020-02-18
NINGBO UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

The most commonly used cross-linking agent is water-soluble glutaraldehyde, which has high reactivity and is difficult to control the dosage. It is easy to cause excessive cross-linking of the enzyme and cause a great loss of enzyme activity. In addition, the tr

Method used

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  • Co-crosslinking immobilization method of beta-galactosidase
  • Co-crosslinking immobilization method of beta-galactosidase
  • Co-crosslinking immobilization method of beta-galactosidase

Examples

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Embodiment Construction

[0015] Enzyme immobilization

[0016] 1) Bisphenol A epoxy resin (brand name is E-44, epoxy value is 0.44, number average molecular weight is 454), three kinds of components of methanol and diethylenetriamine are mixed according to the mass ratio of 2: 2: 1, Stir and react in the range of 25-35°C for 4-5 hours, pour the mixture into water, wash the precipitate repeatedly with water to remove methanol and a small amount of amine, then put it in a vacuum oven and dry it at room temperature to obtain an aminated epoxy resin;

[0017] 2) Add epoxy resin amides and β-cyclodextrin into water at a molar ratio of 1:2.1 to 1:2.3, heat and stir until all epoxy resin amides are converted into molecular complexes and dissolved in water, and keep the The total mass concentration of the aqueous solution is in the range of 5 to 6wt.%.

[0018] 3) dissolving β-galactosidase in a sodium phosphate buffer solution with pH=7.0, and keeping the concentration of the enzyme in the range of 1.0-7.0 ...

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Abstract

The invention relates to a co-crosslinking immobilization method of beta-galactosidase. Oil-soluble diethylene glycol diacrylate is used as a cross-linking agent, a reactant in an aqueous phase is a molecule compound prepared from amino containing beta-galactosidase, aminated epoxy resin and beta-cyclodextrin, through a Michael addition reaction of dual bonds and amino, the co-crosslinking polyreaction is generated at low temperature, and the immobilization beta-galactosidase of different loading quantities can be prepared. The cross-linking degree is controlled, so that the dispersibility isimproved, and internal mass transfer microenvironment can be improved. An immobilized enzyme has high catalytic activity, and when the loading quantity is 56mg of enzymes/1g of carriers, the relativeactivity can achieve the highest value, and is 93% of that of free enzymes.

Description

technical field [0001] The invention relates to the technical field of immobilized enzyme biocatalysis, in particular to a method for co-crosslinking and immobilizing β-galactosidase. The novel immobilized β-galactosidase can be specially used to improve the sweetness of dairy products. It can also reduce the content of lactose, and it is convenient for lactose intolerant consumers to accept milk, a natural and well-balanced food. Background technique [0002] β-galactosidase is an acidic hydrolase (isoelectric point 4.6), with a molecular weight of 540kD and a tetramer structure, widely present in animals, plants and microorganisms. Lactose is a unique sugar in milk. However, children and adults are prone to lack of lactase in their bodies, and drinking milk often causes indigestion and absorption of lactose. If a person with lactase deficiency consumes more lactose at one time, it will cause symptoms such as abdominal distension and diarrhea, which is called lactose into...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/089
CPCC12N11/10C12N11/08C12N9/2471C12Y302/01023Y02P20/50
Inventor 吴嘉沁张瑞丰李艳肖通虎龙能兵
Owner NINGBO UNIV
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