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Preparation method and application of immobilized acid urease for wine

A technology for acid urease and wine, which is applied in the preparation of alcoholic beverages, biochemical equipment and methods, and methods based on microorganisms, etc. It can solve the problems of unstable free enzymes, difficult separation and recovery of enzymes, variability and inactivation, etc. Achieve high urea removal rate, easy recycling, and reduce urea content

Inactive Publication Date: 2011-08-17
宁夏乙征生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Adding wine-used acid urease to wine can reduce the urea content, but the free enzyme is unstable in the solution and is easily denatured and inactivated. After the reaction, the enzyme is difficult to separate and recover, and cannot be reused. Using immobilized enzymes can avoid the above shortcomings and expand The scope and frequency of use of the enzyme

Method used

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  • Preparation method and application of immobilized acid urease for wine
  • Preparation method and application of immobilized acid urease for wine

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Dissolve 2g of sodium alginate in 100mL of free acid urease enzyme solution, stir fully with a glass rod, stir well, place it in a refrigerator at 4°C and refrigerate until there are no bubbles in the solution, drop 500mL of the solution with a syringe. In a solution containing 0.25% chitosan and 5% calcium chloride, place it on a magnetic stirrer for 3 hours, wash it with deionized water, remove free chitosan and calcium chloride, drain, and remove the immobilized small The spheres were placed in 0.1% glutaraldehyde, the mass of the immobilized spheres / volume of glutaraldehyde was controlled to be 2 g / mL, and after 3 min of microwave, it was placed on a magnetic stirrer for magnetic stirring for 2.5 h (covered with an opaque container to prevent glutaraldehyde). The dialdehyde changes color upon seeing light), washed with deionized water to remove free glutaraldehyde (detected by ultraviolet spectrophotometry), drained, refrigerated for later use, to obtain 5...

Embodiment 2

[0046] Example 2: Dissolve 3.6g of sodium alginate in 180mL of free acid urease enzyme solution, stir well with a glass rod, stir well, place it in a refrigerator at 4°C and refrigerate until there are no bubbles in the solution, drop into 900mL dropwise with a syringe Dissolved in a solution of 0.25% chitosan and 5% calcium chloride, placed it on a magnetic stirrer for 3 hours, and washed it with deionized water to remove free chitosan and calcium chloride. The pellets were placed in 0.1% glutaraldehyde, and the mass of the immobilized pellets / volume of glutaraldehyde was controlled to be 2 g / mL. After microwave for 3 min, it was placed on a magnetic stirrer for 2.5 h (covered with an opaque container to prevent Glutaraldehyde changes color upon exposure to light), washed with deionized water to remove free glutaraldehyde (detected by ultraviolet spectrophotometry), drained, refrigerated for later use, to obtain 89.58g of immobilized acid urease for wine, loaded into a flow de...

Embodiment 3

[0047] Example 3: Dissolve 2g of sodium alginate in 100mL of free acid urease enzyme solution, fully stir with a glass rod, stir evenly, place it in a refrigerator at 4°C and refrigerate until there are no bubbles in the solution, drop into 500mL of solution with a syringe dropwise. In a solution containing 0.25% chitosan and 5% calcium chloride, place it on a magnetic stirrer for 3 hours, wash it with deionized water, remove free chitosan and calcium chloride, drain, and remove the immobilized small The spheres were placed in 0.1% glutaraldehyde, the mass of the immobilized spheres / volume of glutaraldehyde was controlled to be 2g / mL, and after microwave for 3min, placed on a magnetic stirrer for 2.5h (covered with an opaque container to prevent glutaraldehyde) The dialdehyde changes color upon exposure to light), washed with deionized water to remove free glutaraldehyde (detected by UV spectrophotometry), drained, refrigerated for later use, to obtain 50.56g of immobilized aci...

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Abstract

A preparation method and application of immobilized acid urease for wine belong to the technical fields of enzyme preparation and brewing additive. The preparation method comprises the steps that purified free enzyme separated from enterobacter 9043C12 is subjected to sodium alginate and calcium chloride embedding, chitosan adsorption and trace glutaraldehyde crosslinking to obtain the immobilized acid urease for the wine; the optimal temperature of the immobilized acid urease for the wine is 40 DEG C, optimal pH thereof is 4, and storage half-life period is 71 days; and the invention also relates to the application of the immobilized acid urease for the wine, after yellow wine is added with the immobilized acid urease for the wine, urea removal rate of the yellow wine can reach about 80 percent, and after the immobilized acid urease for the wine is continuously used for 21 times, the urea removal rate can reach 51.6 percent. The invention can realize repeated use of the immobilized acid urease for the wine, without changing the process conditions of the original brewed wine or the flavor of the wine, thus being a more ideal method for removing urea in the wine.

Description

technical field [0001] A kind of preparation method and application of immobilized acid urease for wine, specifically from Enterobacter 9043C 12 The free enzyme isolated and purified from the medium is embedded with sodium alginate and calcium chloride, adsorbed by chitosan, and cross-linked with a trace amount of glutaraldehyde to obtain immobilized acid urease for wine, which belongs to the technical field of enzyme preparations and wine additives. Background technique [0002] Urethane (or urethane, EC for short) was classified into Group 2B (suspected to be carcinogenic to humans) by the International Agency for Research on Cancer (IARC) in 2005. Its carcinogenic mechanism can be seen through the study of the metabolic pathways of urethane in the body. There are three ways: the first more than 90% of urethane can be decomposed into ethanol, ammonia and carbohydrates, etc. (this pathway is non-toxic. ); the second about 0.5% of urethane is oxidized to DNA addition polyme...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/10C12N11/04C12H1/15C12R1/19
Inventor 田亚平吴召慧吕园园
Owner 宁夏乙征生物工程有限公司
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