A kind of preparation method of mesoporous immobilized cellulase

A cellulase and mesoporous technology, which is applied in the field of materials, can solve the problems of reduced enzyme activity, poor stability, and bioincompatibility, and achieve the goals of improved enzyme stability and catalytic activity, mild production reaction conditions, and reduced use costs. Effect

Inactive Publication Date: 2016-09-07
廖文强
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Enzymes that are not immobilized by mesoporous materials, that is, free enzymes, have the following disadvantages in actual use: (1) cannot be recycled and cannot be reused; (2) have poor stability, and the enzyme activity is reduced or even lost in some environments; (3) Enzyme catalytic efficiency is low
[0012] The current preparation methods of mesoporous materials are not conducive to industrial production, such as the need for high temperature or high pressure; or the need for surfactant glutaraldehyde, polyethylene glycol, polyvinyl alcohol, span80, etc. as templates; or the need to add ethanol or Alcohols such as methanol; or the preparation environment of inert gas such as nitrogen is required; or the preparation time is long; in addition, the mesoporous materials prepared by these methods have a narrow pore size range and a small pore size adjustment range, which makes the preparation process of mesoporous materials complicated and biologically unstable. Compatible, unfriendly to the environment, high production cost, not industrialized

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  • A kind of preparation method of mesoporous immobilized cellulase
  • A kind of preparation method of mesoporous immobilized cellulase
  • A kind of preparation method of mesoporous immobilized cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: preparation of mesoporous material

[0047] Table 2. Raw materials required to produce 400kg of mesoporous materials

[0048]

[0049] The preparation process is as follows:

[0050]Ethyl orthosilicate, water, and hydrochloric acid are stirred and mixed in proportion at room temperature to carry out sol-gel reaction. The reaction time is about 15 minutes to obtain a homogeneous colorless and transparent sol, which is cooled with water to fully remove and recover the reaction product. The by-product ethanol forms a pre-hydrolyzed gel, then adds D-glucose or fructose aqueous solution under stirring, and finally adds sodium hydroxide solution to adjust the pH value of the system to near neutral, and then place it at low temperature to form a gel for about 48 hours After the gelation is complete, crush the gel, wash it with water for 2-3 times to remove the glucose template in the gel system, filter and dry the mesoporous material for 2-3 days, and further...

Embodiment 2

[0055] Embodiment 2: Preparation of the mesoporous immobilized enzyme of the present invention

[0056] See Table 3 for the preparation of mesoporous immobilized cellulase as an example.

[0057] table 3

[0058]

[0059] When preparing mesoporous immobilized cellulase, the mass ratio of mesoporous material to enzyme is 4:2.

[0060] One-step preparation of mesoporous immobilized enzymes:

[0061] Referring to Example 1, the step of preparing a mesoporous material to form a pre-hydrolyzed gel, after adding D-glucose aqueous solution under stirring, then adding cellulase buffer in proportion under stirring, and finally adding sodium hydroxide solution to adjust The pH value of the system is near neutral, and then placed in a low temperature environment to form a gel. After the gelation is completed for about 48 hours, the obtained wet gel is ground and pulverized, and washed with water twice to remove the glucose template in the gel system. Filter, dry for 2-3 days, grind...

Embodiment 3

[0062] Embodiment 3: Immobilized enzyme stability and catalytic efficiency are improved

[0063] After immobilizing cellulase with the prepared mesoporous material, the thermal stability is good, and 50% fructose is the template with the highest thermal stability, which is obviously better than immobilized enzyme and free enzyme without template, and cellobiose in free cellulase The enzyme activity is around 256IU (37°C), as shown in Table 4 and Table 5:

[0064] Table 4 immobilized cellulase has good thermal stability

[0065]

[0066] Note: (1) Cellulase is a complex enzyme composed of C1 enzyme, Cx enzyme and β-glucosidase (ie cellobiase).

[0067] (2) The enzyme activity curves in Table 4 take the enzyme activity of each enzyme at 37°C as 1. At 37°C, the measured enzyme activity: cellobiose in 0% fructose template mesoporous immobilized cellulase The enzyme activity is 105±8IU, the cellobiase activity in 50% fructose template nano-cellulase is 256±13IU, and the disacc...

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Abstract

The invention relates to the field of materials and discloses a preparation method of a mesoporous material. The preparation method comprises the following steps: adding an acid or alkali solution into a precursor in a stirring state, performing hydrolysis to obtain a transparent solution, cooling the transparent solution and recycling ethyl alcohol; adding a template solution into the transparent solution in a stirring state, and adjusting the pH value of the mixed solution to be neutral by an acid or alkali solution for gelatinizing; smashing wet glue obtained by the gelatinizing; removing the template through water washing; carrying out suction filtration on the mixed solution after water washing to obtain white gel; and drying the gel, smashing, and sieving, thus obtaining the mesoporous material. According to the preparation method of the mesoporous material provided by the invention, an immobilized enzyme can be obtained by adding bioactive substances including enzymes into a reaction system in the preparation process. The experiment shows that the enzyme activity of the immobilized enzyme of the mesoporous material nearly keeps unchanged after the immobilized enzyme is repeatedly used.

Description

technical field [0001] The invention relates to the field of materials, in particular to a preparation method of mesoporous immobilized cellulase. Background technique [0002] Under the dual pressure of increasing demand for crude oil and environmental pollution in my country, the development of renewable clean energy is of great significance for reducing dependence on crude oil and alleviating environmental pressures such as "smog". In recent years, significant progress has been made in the production of bio-liquid fuels from biomass. The current technology and technology have been able to use cellulose-containing biomass that is not directly edible by humans, such as straw and bagasse, as raw materials to prepare cellulosic ethanol. [0003] The production of ethanol from biomass generally involves the following steps: [0004] (1) Collection and transportation of biomass raw materials; [0005] (2) Biomass pretreatment; [0006] (3) Using cellulase to decompose and c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J13/00C01B33/14C12N11/04
Inventor 廖文强张翼冠
Owner 廖文强
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