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A method for immobilizing cellulase and hydrolyzing cellulose

A technology of cellulase and cellulose, applied in biochemical equipment and methods, fixed on/in organic carriers, enzymes, etc., can solve problems such as increased steric hindrance, unfavorable cellulase hydrolysis, etc., to achieve Reduce the Michaelis constant, improve the degradation rate of TS, and improve the effect of recycling

Active Publication Date: 2021-11-26
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reason is that the movement of enzyme molecules is limited after immobilization, which increases the steric hindrance between the enzyme molecule and the substrate, which is not conducive to the effective hydrolysis of cellulase.

Method used

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  • A method for immobilizing cellulase and hydrolyzing cellulose
  • A method for immobilizing cellulase and hydrolyzing cellulose
  • A method for immobilizing cellulase and hydrolyzing cellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Method for cellulase immobilization using sodium alginate-polyethylene glycol-chitosan

[0027] 1) Accurately weigh the cellulase powder and dissolve it in pure water to make the concentration 5mg / ml, set aside.

[0028] 2) Accurately weigh 1.5g chitosan in a 100ml beaker, add 48.5g mass fraction of 2% acetic acid to make the mass fraction of chitosan 3%, after mixing uniformly at room temperature, form a chitosan solution for use .

[0029] 3) Accurately weigh glutaraldehyde to be configured into a 1% glutaraldehyde solution by mass fraction, and set aside;

[0030] 4) Immobilization of cellulase: Accurately weigh 2.625g of polyethylene glycol in a 100ml beaker, add 48.25g of 0.05mol / L citric acid-sodium citrate buffer solution with pH=4.8, stir with a glass rod at room temperature After dissolving, 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 ...

Embodiment 2

[0037] The preparation of sodium alginate-polyethylene glycol-chitosan immobilized cellulase, operation is the same as embodiment 1, fixation condition gets respectively the lowest value and the highest value described in "summary of the invention" of each factor, to immobilization The enzyme conditions were optimized and analyzed, and the results are shown in Table 2.

[0038] Table 2 Condition optimization analysis of immobilized cellulase

[0039]

[0040]

Embodiment 3

[0042] Sodium alginate-polyethylene glycol-chitosan immobilized cellulase for hydrolysis of microcrystalline cellulose

[0043] 1) Sodium alginate-polyethylene glycol-chitosan immobilized cellulase enzymatically hydrolyzes microcrystalline cellulose: a 50 ml shake flask is used. 0.5 g of TS microcrystalline cellulose was added, the dosage of immobilized enzyme was 1.745 g, and a citric acid-sodium citrate buffer solution with pH=4.93 was added to make the substrate mass fraction 6.89%. Parafilm seals to prevent moisture from evaporating. Enzymatically hydrolyze at 60°C and 150r / min for 48h, suction filter after the reaction, measure the reducing sugar content in the filtrate to obtain the enzymatic hydrolysis yield; after drying the solid residue, measure the mass change before and after the reaction.

[0044] 2) In order to illustrate the advantages of the immobilization method of the present invention, use sodium alginate immobilized cellulase to enzymatically hydrolyze mic...

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Abstract

The invention provides a method for immobilizing cellulase and hydrolyzing cellulose. The method for preparing immobilized cellulase uses sodium alginate as a carrier, and utilizes OH in sodium alginate ‑ and COOH ‑ Oxygen atom and Ca 2+ The principle of ion exchange reaction forms a gel, polyethylene glycol is used as a porogen and a hydrophilic agent, glutaraldehyde is used as a crosslinking agent, and the gel is formed between chitosan and sodium alginate-polyethylene glycol immobilized enzyme The polyelectrolyte membrane improves the stability of the immobilized enzyme. Using immobilized cellulase to hydrolyze cellulose can realize the repeated utilization of cellulase. Adding polyethylene glycol and chitosan can improve the enzyme fixation rate and enzyme activity of immobilized cellulase, reduce the Michaelis constant, increase the pore density of the carrier surface, increase the reducing sugar content of the enzymatic hydrolysis reaction, the enzymatic hydrolysis yield and TS degradation rate. The preparation and application method of the invention is simple and convenient to operate, mild in fixing conditions, low in investment and friendly to the environment.

Description

technical field [0001] The invention specifically relates to a method for immobilizing cellulase and a method for hydrolyzing cellulose with 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. Due to its high crystallinity and good hydrogen bonds between molecules a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/10C12N1/08C12P19/14C12P19/04
CPCC12N1/08C12N9/2437C12N11/10C12P19/04C12P19/14
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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