Method for preparing immobilized cellulase

A cellulase and product technology, applied in the field of immobilized cellulase prepared by physical adsorption, can solve the problems of enzyme structure change, achieve stable properties, increase aroma components, and reduce nicotine effects

Pending Publication Date: 2019-07-23
CHINA TOBACCO YUNNAN IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although many preparation methods of immobilized cellulase have appeared successively, most of them have applied cross-linking agents or other systems that can react with enzymes, resulting in unavoidable changes in the enzyme structure during the cross-linking process, such as (Pei J et al. al. ANovel Layered Anchoring Structure Immobilized Cellulase via Covalent Binding of Cellulase on MNPs Anchored by LDHs. Journal of Inorganic and Organometallic Polymers and Materials, 2018), therefore, it is extremely important to develop a simple and fast immobilized cellulase technology

Method used

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  • Method for preparing immobilized cellulase
  • Method for preparing immobilized cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Preparation of immobilized cellulase: Weigh 100 mg of UiO-66 MOFs material into a test tube, add 100 mg of cellulase and 10 mL of water, mix well, place it in a 30°C water bath for oscillating adsorption for 5 hours, and obtain an oscillating product. Centrifuge the oscillating product twice, collect and combine the supernatant to obtain the centrifuged supernatant, wash the centrifuged precipitate three times with ultrapure water to obtain the centrifuged product, and dry the centrifuged product in a 30°C oven to obtain immobilized cellulase sample 1 .

[0036] The centrifuged supernatant was retained for measuring the cellulase loading rate, and the immobilized cellulase sample 1 was used for measuring the enzyme activity of the cellulase.

[0037] Determination of the loading rate of immobilized cellulase: Add 1mL of centrifuged supernatant and 1mL of water to two test tubes respectively, then add 5mL of staining solution to the test tubes, mix well, and wait for 5 m...

Embodiment 2

[0040] The preparation method of the immobilized cellulase was the same as that in Example 1, but the adsorption time was changed from 5 hours of adsorption in Example 1 to 7 hours of adsorption to obtain immobilized cellulase sample 2. The loading rate of immobilized cellulase and the enzyme activity of immobilized cellulase were measured under the same conditions.

Embodiment 3

[0042] The preparation method of the immobilized cellulase was the same as in Example 1, except that the adsorption time was changed, and the adsorption time in Example 1 was changed from 5 h to 9 h, and immobilized cellulase sample 3 was obtained. The other things were unchanged, and the loading rate of immobilized cellulase was measured under the same conditions, and the enzyme activity of immobilized cellulase was measured.

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Abstract

The invention discloses a method for preparing immobilized cellulase. Cellulase serving as a raw material is immobilized by using UiO-66MOFs material as a carrier. The method comprises the following steps: (1) dissolving 50-100mg of cellulase in ultrapure water, adding 50-100mg of a UiO-66MOFs material into the mixture, performing oscillation in a thermostatic water-bath at 20-40 DEG C for 8-20 hours to obtain an oscillation product; (2) centrifuging the oscillation product in the step (1) to obtain centrifugal supernate and a centrifugal product; and (3) drying the centrifugal product in thestep (2) to obtain the immobilized cellulase. The method has the advantages of high enzyme loading and high activity recovery rate. The immobilized cellulase can be used for effectively degrading cellulose macromolecules in reconstituted tobaccos when being applied to raw material pre-treatment of reconstituted tobaccos, and main aroma components are added to reduce nicotine and improve the aromaand harmony of the reconstituted tobaccos.

Description

technical field [0001] The invention relates to the preparation of immobilized cellulase by physical adsorption method, in particular to a preparation method of immobilized cellulase. Background technique [0002] With the increasing depletion of energy, more and more people's attention is focused on renewable energy, and cellulose is a renewable energy and the energy with the largest reserve in the world. The development and utilization of cellulose can greatly reduce the current panic about energy shortages. The author Meng C reported a method for utilizing cellulose (Meng C et al. Activation of Cellulose by Supercritical Tetrafluoroethane and Its Application in Synthesis of Cellulose Acetate. Industrial & Engineering Chemistry Research, 2015 ,54(48)). Due to the special structure of cellulose, the decomposition of cellulose is more difficult than other polysaccharides. Although there are some reports on the degradation of cellulose, the conditions are harsh, such as the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/02
CPCC12N11/14C12N11/02C12N9/2437
Inventor 韦克毅张玲汤丹瑜杨建云张天栋王家强刘晓敏朱龙凌军
Owner CHINA TOBACCO YUNNAN IND
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