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Method for immobilizing mannase by taking macroporous resin as carrier and application of immobilized mannase

A technology of mannanase and macroporous resin, applied in the direction of immobilization on/in organic carrier, fermentation, etc., can solve difficult industrial production and practical application, low efficiency of mannanase immobilization, processing technology Complicated problems, to achieve significant degradation effect, stable properties, and simple processing technology

Inactive Publication Date: 2012-07-18
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the common defects of these two methods are: low mannanase immobilization efficiency, complex processing technology, high production cost, unstable properties and other defects, which are difficult to industrial production and practical application

Method used

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  • Method for immobilizing mannase by taking macroporous resin as carrier and application of immobilized mannase
  • Method for immobilizing mannase by taking macroporous resin as carrier and application of immobilized mannase
  • Method for immobilizing mannase by taking macroporous resin as carrier and application of immobilized mannase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: AB-8 macroporous adsorption resin is used as carrier

[0029] (1) Soak the AB-8 macroporous adsorption resin with 95% ethanol for 12 hours, and stir it from time to time to make it fully swell, rinse the resin with 95% ethanol until the ethanol solution is mixed with water in a ratio of 1:5 and does not appear Until the white turbidity, finally rinse with deionized water repeatedly until the washing liquid has no ethanol smell;

[0030] (2) Put the treated resin into 500 mL of acetic acid-sodium acetate buffer solution containing 3000 U of mannanase, the pH value of the buffer solution is 5.5, and keep the temperature at 45° C. for 8 hours for oscillating adsorption.

[0031] (3) Using glutaraldehyde with a mass concentration of 4%, oscillating and cross-linking for 3 hours, vacuum filtration to obtain immobilized mannanase, and storing in a refrigerator at 4°C for later use.

Embodiment 2

[0032] Embodiment 2: X-5 macroporous adsorption resin is made carrier

[0033] (1) Soak X-5 macroporous adsorption resin with ethanol overnight, wash with distilled water for several times, then soak in distilled water for several times until neutral, soak in distilled water for several times until neutral, soak more than double the volume of distilled water and place in a 4°C refrigerator spare.

[0034] (2) Put the treated resin into 500 mL of acetic acid-sodium acetate buffer solution containing 3000 U of mannanase, the pH value of the buffer solution is 5.5, and keep the temperature at 45° C. for 8 hours for oscillating adsorption.

[0035] (3) Using glutaraldehyde with a mass concentration of 4%, oscillating and crosslinking for 3 hours, collecting the filter cake by vacuum filtration, obtaining immobilized mannanase by vacuum filtration, and storing it in a refrigerator at 4°C for later use.

Embodiment 3

[0036] Embodiment 3: D-151, D-152 cation exchange resin carrier

[0037] (1) Take a certain amount of D-151 and D-152 cation exchange resins and soak them in distilled water to swell and remove impurities, then treat them with 0.1mol / L HCl for 1 hour, wash them with distilled water until they are neutral, and then treat them with 0.1mol / L NaOH After 1 hour, wash with distilled water until neutral, blot dry, and store in a 4°C refrigerator for later use.

[0038] (2) Put the treated resin into 500 mL of acetic acid-sodium acetate buffer solution containing 3000 U of mannanase, the pH value of the buffer solution is 5.5, and keep the temperature at 45° C. for 8 hours for oscillating adsorption.

[0039] (3) Using glutaraldehyde with a mass concentration of 4%, oscillating and crosslinking for 3 hours, collecting the filter cake by vacuum filtration, obtaining immobilized mannanase by vacuum filtration, and storing it in a refrigerator at 4°C for later use.

[0040] The immobili...

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Abstract

The invention discloses a method for immobilizing mannase by taking a macroporous resin as a carrier and application of immobilized mannase. The invention is characterized in that the method for immobilizing the mannase comprises the following steps of: (1) soaking the macroporous resin in ethanol for 6 to 12 hours, filtering out the ethanol, washing until residue is not produced after an eluent is evaporated to dryness, drying the macroporous resin cleaned by using the ethanol, and preserving for later use; (2) putting the dried macroporous resin into an acetic acid-sodium acetate buffer solution containing the mannase, keeping the temperature at 45 to 60 DEG C, and performing oscillation adsorption for 6 to 12 hours, wherein the pH value of the buffer solution is 4.0 to 6.0; and (3) performing oscillation cross-linking for 2 to 6 hours by using glutaraldehyde at the mass concentration of 0.5 to 4 percent, suction-filtering under vacuum to obtain the immobilized mannose, putting into a refrigerator at the temperature of 4 DEG C, and storing. The invention has the advantages that: the process is simple, and the immobilized mannase is low in cost, has stable properties and can be repeatedly used; and the immobilized mannose has a more remarkable effect of degrading aloe polysaccharides containing mannan polysaccharides.

Description

technical field [0001] The invention relates to the technical field of enzyme immobilization, in particular to a mannanase immobilization method using a macroporous resin as a carrier, and the application of the immobilized mannanase in the degradation of aloe polysaccharides. Background technique [0002] β-mannanase is a kind of endonuclease that can hydrolyze mannan oligosaccharides and mannan polysaccharides containing β-mannosidic bonds, and belongs to hemicellulase. β-mannanase is widely used in food, textile and petroleum industries. In recent years, the research on β-mannanase has gradually become a research hotspot at home and abroad. In the process of applying mannanase, people have found some shortcomings of free mannanase, such as poor stability, free mannanase is easy to lose activity under the influence of temperature, pH value, inorganic ions and other factors; The free mannanase after the enzymolysis reaction with the substrate in water is difficult to sepa...

Claims

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Application Information

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IPC IPC(8): C12N11/02C12P19/14
Inventor 梁振益孙达远林栖凤黄小周
Owner HAINAN UNIVERSITY
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