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Cellulase immobilization and cellulose hydrolysis methods

A technology of cellulase and cellulose, applied to 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 The effect of reducing the Michaelis constant, increasing the degradation rate of TS, and increasing the fixation rate of enzyme activity

Active Publication Date: 2018-04-10
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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  • Cellulase immobilization and cellulose hydrolysis methods
  • Cellulase immobilization and cellulose hydrolysis methods
  • Cellulase immobilization and cellulose hydrolysis methods

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

Cellulase immobilization and cellulose hydrolysis methods are provided. According to the cellulase immobilization method, sodium alginate is adopted as a carrier, a principle that an ion exchange reaction occurs between oxygen atoms in OH- and COOH- in the sodium alginate and Ca<2+> is utilized to form gel, polyethylene glycol is adopted as a pore-forming agent and a hydrophilic agent, glutaraldehyde is adopted as a crosslinking agent, and a polyelectrolyte membrane formed between chitosan and the sodium alginate-polyethylene glycol immobilized enzyme is utilized to improve stability of the immobilized enzyme. The immobilized cellulase is used for cellulose hydrolysis, and repeated utilization of the cellulase can be achieved. Through addition of the polyethylene glycol and the chitosan, the enzyme activity immobilization rate and enzyme activity of the immobilized cellulase are improved, the Michaelis constant is reduced, pore density of a carrier surface is increased, and the reducing sugar content of an enzyme hydrolysis reaction, the enzyme hydrolysis yield and the TS degradation rate are increased. The preparing and application methods are simple to operate, immobilization conditions are mild, investment is low, and the methods are environmentally friendly.

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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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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