Co-crosslinking immobilization method for acid urease

An acid urease, co-crosslinking technology, applied in immobilized enzymes, biochemical equipment and methods, immobilized on/in organic carriers, etc., can solve the problem of high reactivity, excessive crosslinking of enzymes, loss of enzyme activity, etc. problem, to achieve the effect of improving the catalytic activity and improving the microenvironment

Active Publication Date: 2020-02-11
NINGBO UNIV
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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 traditional cross-linking method often It is necessary to precipitate and aggregate enzyme molecules before cross-linking, which will not only cause waste of enzymes, but also block mass transfer channels, and cannot fully exert the catalytic efficiency of enzymes

Method used

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  • Co-crosslinking immobilization method for acid urease
  • Co-crosslinking immobilization method for acid urease
  • Co-crosslinking immobilization method for acid urease

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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 triethylenetetramine are mixed according to the mass ratio of 2: 2: 1.2, 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 acid urease in a sodium phosphate buffer solution with pH=7.5, keeping the concentration of the enzyme in the range of 1.0-7.0 mg / m...

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Abstract

The invention relates to a co-crosslinking immobilization method for acid urease. Oil-soluble butanediol diacrylate is used as a cross-linking agent; reactant in an aqueous phase is amino-containing acid urease and a supramolecular complex formed by aminated epoxy resin and beta-cyclodextrin; and through the utilization of the Michael addition reaction of double bonds and amino groups, co-crosslinking polymerization can occur under a low temperature, and immobilized acid urease with different loading amount can be prepared. Through the controlling of the degree of cross-linking, dispersibilitycan be enhanced, internal mass transfer microenvironment can be improved; and the immobilized enzyme can have high catalytic activity when the loading amount is in the range of 21-58 mg enzyme/g carrier, and the highest relative activity can reach 90% 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 acid urease. The novel immobilized acid urease can be specially used for removing urea in wine. Background technique [0002] Urease (EC 3.5.1.5), also known as aminohydrolase, is a metalloenzyme containing nickel ions (the isoelectric point is 4.6-4.8). Its molecular weight is about 120,000 to 130,000 Da. In the presence of water, urea is decomposed into ammonia and carbon dioxide, 10 times the speed of this reaction without urease catalysis. According to the optimum pH of urease, it can be divided into acid urease, neutral urease and alkaline urease. Urease is widely distributed in the seeds of plants, and the content is most abundant in soybean and sword bean. In addition, it also exists in animal blood and urine, and some microorganisms can also secrete urease. In the wine fermentation industry, when th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/089C12N9/80
CPCC12N11/10C12N11/08C12N9/80C12Y305/01005Y02P20/50
Inventor 吴嘉沁张瑞丰李艳肖通虎龙能兵
Owner NINGBO UNIV
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