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Preparation method and application of immobilized acidic urease membrane

A technology of acid urease enzyme film and acid urease, which is applied to the preparation of alcoholic beverages, fixed on/in organic carriers, hydrolytic enzymes, etc., can solve the problems of difficult long-term storage, application restrictions, and difficult reuse, and achieve High urea removal rate, easy recycling, and the effect of reducing urea content

Inactive Publication Date: 2011-05-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The application of free acid urease is limited due to its disadvantages such as difficulty in maintaining activity, difficulty in reutilization and difficulty in long-term storage.

Method used

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  • Preparation method and application of immobilized acidic urease membrane
  • Preparation method and application of immobilized acidic urease membrane
  • Preparation method and application of immobilized acidic urease membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Dissolve 4 mL of acid urease in 100 mL of 0.8% chitosan solution, stir well with a glass rod, and place in a refrigerator at 4°C until there are no bubbles in the solution. 2 amount

[0047] Pour into a smooth glass plate, dry at 37°C to form a film, soak in a small amount of citric acid buffer solution, peel off, drain, cut the film into squares of uniform size, place in 0.02% glutaraldehyde solution, and place under magnetic force Magnetically stir on the stirrer for 2 hours (cover with an opaque container to prevent glutaraldehyde from changing color when exposed to light), wash with deionized water to remove free glutaraldehyde (detected by UV spectrophotometry), drain, refrigerate for later use, and immobilize Acid urease enzymatic membrane. Take a 10 cm×10 cm membrane, put it into 100mL rice wine, and act for 24 hours at 37°C to measure the urea content in the rice wine and calculate the urea removal rate. rate reached 73%.

Embodiment 2

[0048] Example 2: Dissolve 4 mL of acid urease in 100 mL of 0.8% chitosan solution, stir thoroughly with a glass rod, and place in a refrigerator at 4°C until there are no bubbles in the solution. 2 Pour the amount into a smooth glass plate, dry at 37°C to form a film, soak in a small amount of citric acid buffer solution, peel off, drain, cut the film into squares of uniform size, place in 0.02% glutaraldehyde solution, and place Stir magnetically on a magnetic stirrer for 2 hours (cover with an opaque container to prevent glutaraldehyde from changing color when exposed to light), wash with deionized water to remove free glutaraldehyde (detected by ultraviolet spectrophotometry), drain, and refrigerate for later use to obtain Immobilized acid urease enzyme membrane. Take a 4 cm × 4 cm membrane, put it into 100 mL of rice wine, and let it act for 24 hours at 37 ° C to measure the urea content and calculate the urea removal rate. The urea content dropped from 24.8mg / L to 9.6mg...

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Abstract

The invention discloses a preparation method and application of an immobilized acidic urease membrane, belonging to the technical field of enzymic preparations and brewing additives. The preparation method comprises the steps of: dissolving acidic urease which is separated and purified from enterobacter 9043C12 in a certain amount of chitosan solution; pouring the solution in a smooth glass plate; naturally dehydrating and drying into a membrane at a temperature of 37DEG C; and crosslinking by using micro amount of glutaraldehyde so as to obtain the immobilized acidic urease membrane, wherein the immobilized acidic urease membrane has the optimum temperature of 40DEG C, the optimum pH of 4.5, the storage half-time of 40d and the enzyme activity recovery of 64.3%. The application of the immobilized acidic urease membrane is that the urea removal rate can achieve 73% after the membrane is added in yellow rice wine and still can achieve 50% after 10-times continuous use. A product of the immobilized acidic urease membrane takes a membrane as a fixed carrier which has a large contact area and can be repeatedly used; and when the immobilized acidic urease membrane is used in the yellow rice wine, both the conditions of the original brewing process and the flavor of the wine are not changed and the urea in the wine can be removed more ideally.

Description

technical field [0001] Preparation method and application of immobilized acid urease membrane, specifically from enterobacter 9043C 12 Acid urease was obtained by separating and purifying from medium; dissolving acid urease in a certain amount of chitosan solution, pouring it into a smooth glass plate, drying and dehydrating naturally at 37°C to form a film, cross-linking with a small amount of glutaraldehyde, and obtaining immobilized acid urease An enzyme film belongs to the technical field of enzyme preparations and brewing additives. Background technique [0002] Acid urease is an enzyme that can decompose urea (urea) into ammonia and carbon dioxide or ammonium carbonate. The enzyme is widely distributed in the seeds of plants, but it is abundant in soybean and sword bean. It also exists in animal blood, urine and microorganisms. It can withstand acidic environment and is still active in the system of pH 4.0~5.5. Urea is a by-product produced in the process of microb...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N9/80C12G3/08
Inventor 田亚平吕园园
Owner JIANGNAN UNIV
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