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Method for improving activity of immobilized cellulase

A technology of cellulase and activity, which is applied in the field of bioengineering, can solve problems such as the reduction of immobilized enzyme activity, and achieve the effect of improving enzyme activity and improving activity

Inactive Publication Date: 2013-04-17
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the immobilized enzyme can be reused, the activity of the immobilized enzyme is reduced because some enzyme molecules are embedded in the carrier, so the research on improving the activity of the immobilized enzyme is a hot spot in current research.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Preparation of immobilized cellulase: accurately measure chitosan and the volume fraction of 2% acetic acid solution is 1: 100g / mL, stir continuously with a glass rod, finally dissolve into a transparent colloidal solution, take the above-mentioned certain amount Chitosan colloidal solution, add the NaOH solution that the mass fraction of 3:4 is 5% with the volume ratio of colloidal solution, obtain flocculant matter, centrifuge and obtain precipitation, wash and precipitate to neutrality, then add and colloidal solution volume ratio 5% glutaraldehyde aqueous solution with a volume fraction of 1:2, stirred with a constant temperature water bath stirrer for 2 hours at room temperature, allowed to stand overnight, discarded the supernatant by centrifugation, washed 3-4 times with water, removed residual glutaraldehyde, centrifuged Obtain the chitosan solid after the cross-linking; In the chitosan after the above-mentioned cross-linking, add the diluted 100 times liquid...

Embodiment 2

[0021] (1) Preparation of immobilized cellulase: accurately measure chitosan and the volume fraction of 2% acetic acid solution is 1: 100g / mL, stir continuously with a glass rod, finally dissolve into a transparent colloidal solution, take the above-mentioned certain amount Chitosan colloidal solution, add the NaOH solution that the mass fraction of 3:4 is 5% with the volume ratio of colloidal solution, obtain flocculant matter, centrifuge and obtain precipitation, wash and precipitate to neutrality, then add and colloidal solution volume ratio 5% glutaraldehyde aqueous solution with a volume fraction of 1:2, stirred with a constant temperature water bath stirrer for 2 hours at room temperature, allowed to stand overnight, discarded the supernatant by centrifugation, washed 3-4 times with water, removed residual glutaraldehyde, centrifuged Obtain the chitosan solid after the cross-linking; In the chitosan after the above-mentioned cross-linking, add the diluted 100 times liquid...

Embodiment 3

[0025] (1) Preparation of immobilized cellulase: accurately measure chitosan and the volume fraction of 2% acetic acid solution is 1: 100g / mL, stir continuously with a glass rod, finally dissolve into a transparent colloidal solution, take the above-mentioned certain amount Chitosan colloidal solution, add the NaOH solution that the mass fraction of 3:4 is 5% with the volume ratio of colloidal solution, obtain flocculant matter, centrifuge and obtain precipitation, wash and precipitate to neutrality, then add and colloidal solution volume ratio 5% glutaraldehyde aqueous solution with a volume fraction of 1:2, stirred with a constant temperature water bath stirrer for 2 hours at room temperature, allowed to stand overnight, discarded the supernatant by centrifugation, washed 3-4 times with water, removed residual glutaraldehyde, centrifuged Obtain the chitosan solid after the cross-linking; In the chitosan after the above-mentioned cross-linking, add the diluted 100 times liquid...

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Abstract

The invention relates to a method for ultrasonically improving the activity of immobilized cellulase, comprising the following steps of: (1) preparation of the immobilized cellulase: accurately weighing and dissolving chitosan and 2% by volume of acetic acid water solutions into transparent colloidal solution, adding 5% by weight of NaOH water solution to a certain amount of the transparent colloidal solution to obtain a flocculent substance, and then adding 5% by volume of glutaraldehyde water solution to obtain crosslinked chitosan; and adding cellulase to the crosslinked chitosan, washing the mixture 3-4 times in water in the second day to remove free enzyme, and then freeze-drying to obtain the immobilized cellulase; and (2) ultrasonic preprocessing of the immobilized cellulase: processing the immobilized cellulase by adopting an ultrasonic generator for 5-30 min, wherein the power of the ultrasonic generator is 10-80 W, and the frequency is 18-29 KHz. The method greatly improves the activity of the immobilized cellulase by combining an enzymologic bioengineering technique with an ultrasonic technique.

Description

technical field [0001] The invention relates to the influence of ultrasonic waves on the activity of immobilized enzymes, belonging to the technical field of bioengineering. Background technique [0002] Cellulose is a fibrous, insoluble crystalline polysaccharide composed of β-1,4-D-glucopyranose residues. It is the main structural material of plant cell walls and the most abundant polymer on earth. Making full use of cellulose can provide an important way for human beings to solve food, environment and energy crises. Under the action of cellulase, cellulose can be decomposed into small molecule cellulose, cellooligosaccharides, cellobiose and glucose which are easily fermented into acetic acid, lactic acid and ethanol. [0003] At present, the enzymatic hydrolysis technology of cellulose has been basically mature. However, in the production process, the high price and large amount of cellulase lead to high production costs and hinder the transformation of this technology...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N9/42C12N13/00
Inventor 王振斌张杰王世清马海乐江慎华马晓珂陈晓寅任晓锋
Owner JIANGSU UNIV
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