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Method for preparing immobilized cellulase and method for hydrolyzing cellulose through immobilized cellulase

A technology of cellulase and cellulose, applied to biochemical equipment and methods, fixed on/in organic carriers, hydrolytic enzymes, etc., can solve problems such as increased steric hindrance, and achieve reduction of Michaelis constant, Improve the immobilization rate of enzyme activity and the effect of increasing the hole density

Pending Publication Date: 2018-05-08
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limited movement of enzyme molecules after immobilization, the steric hindrance between the enzyme molecule and the substrate increases.

Method used

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  • Method for preparing immobilized cellulase and method for hydrolyzing cellulose through immobilized cellulase
  • Method for preparing immobilized cellulase and method for hydrolyzing cellulose through immobilized cellulase
  • Method for preparing immobilized cellulase and method for hydrolyzing cellulose through immobilized cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) Accurately weigh the cellulase powder and dissolve it in pure water to make the concentration 5mg / ml, set aside. Cellulase is produced from Trichoderma viride.

[0026] 2) Preparation of immobilized cellulase: Accurately weigh 2.625 g of polyethylene glycol with a molecular weight of 4000 in a 100 ml beaker, add 48.25 g of 0.05 mol / L citric acid-sodium citrate buffer solution with pH=4.8, and heat at room temperature After stirring and dissolving with a glass rod, 1.75 g of sodium alginate was added so that the mass fraction of sodium alginate was 3.5%, and the mass ratio of sodium alginate to polyethylene glycol was 1:1.5. After mixing evenly at room temperature, a sodium alginate-polyethylene glycol solution is formed, which is ready for use.

[0027] Use a 10ml syringe to draw 5ml of the above-mentioned sodium alginate-polyethylene glycol solution into a 20ml glass bottle, add 1ml of 5mg / ml cellulase solution and 1.5ml of glutaraldehyde solution with a mass fract...

Embodiment 2

[0037] The preparation of sodium alginate-polyethylene glycol immobilized cellulase, operation is the same as embodiment 1 (1) (2), and fixation condition is respectively taken the lowest value and the highest value described in "Summary of the invention (1)" of each factor Value, the condition of immobilized enzyme was optimized and analyzed, and the results are shown in Table 3.

[0038] Sodium alginate-polyethylene glycol immobilized cellulase is used to hydrolyze microcrystalline cellulose, and the operation is the same as in Example 1 (3) (4). The enzymatic hydrolysis conditions were respectively taken the lowest value and the highest value of each factor described in "Summary of the Invention (2)", and the conditions of the enzymatic hydrolysis reaction were optimized and analyzed, as shown in Table 4.

[0039] Table 3 Condition optimization analysis of immobilized cellulase

[0040]

[0041]

[0042] Table 4 Optimal analysis of reaction conditions for enzymatic h...

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Abstract

The invention provides a method for preparing immobilized cellulase and a method for hydrolyzing cellulose through the immobilized cellulase. The method for preparing immobilized cellulase is a crosslinked embedding method, sodium alginate serves as a carrier, gel is formed according to the principle that a ion exchange reaction happens between oxygen atoms of OH- and COOH- in sodium alginate andCa2+, glutaraldehyde serves as a crosslinking agent, polyethylene glycol serves as a pore-foaming agent and a hydrophilic agent, and by adding polyethylene glycol, the enzyme activity immobility and enzyme activity of the immobilized cellulase can be improved, the Michaelis constant is reduced, the pore density of the carrier surface is improved, and the reduction sugar content, enzyme hydrolysisyield and TS degradation rate of enzyme hydrolysis reaction are increased. In the process of hydrolyzing cellulose through the immobilized cellulase, the cellulase can be repeatedly used multiple times. The preparing and applying methods are easy to operate, mild in immobilizing conditions, low in investment and friendly to the environment.

Description

technical field [0001] The invention specifically relates to a preparation method of immobilized cellulase and a method for hydrolyzing cellulose by immobilized cellulase. Background technique [0002] Cellulose is the most widely distributed and abundant natural polymer in nature, and it is the basic material that constitutes the cell wall. Every year, plants produce hundreds of billions of tons of cellulose through photosynthesis, which is an inexhaustible renewable resource. In recent years, with the gradual depletion of coal, oil, natural gas and other non-renewable resources and the increasingly serious environmental problems, the research on the utilization of cellulose as a renewable resource has become one of the frontier fields of international scientific research. [0003] Cellulose is a long-chain unbranched homogeneous polysaccharide formed by β-D-glucopyranose linked by (1-4)-β-glycosidic bonds. The molecular chain has a C1 chair conformation. The three hydro...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/08C12N11/04
CPCC12N9/2437C12N11/04C12N11/08C12N11/10C12Y302/01004
Inventor 刘研萍马一方兰淼李秀金袁海荣
Owner BEIJING UNIV OF CHEM TECH
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