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Preparation method of modified nano-hydroxyapatite carried immobilized cellulase

A nano-hydroxyapatite, cellulase technology, applied in biochemical equipment and methods, glycosylase, immobilized on or in inorganic carriers, etc., can solve the problem of high brittleness and poor reusability of immobilized enzymes , slow degradation speed and other problems, to achieve the effect of high brittleness, good chemical stability and slow degradation speed

Inactive Publication Date: 2017-12-08
苏州凯邦生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And inorganic nano-hydroxyapatite has good biocompatibility, but its brittleness is large, the degradation rate is slow, and the immobilized enzyme made with it has poor reusability, which limits its application in the immobilized preparation of biological enzyme preparations.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A preparation method of immobilized cellulase using modified nano-hydroxyapatite as a carrier, comprising the following steps:

[0028] (1) Mix sodium alginate, water-soluble chitosan and deionized water, and stir at 30°C±5°C until the solid dissolves to obtain a mixed solution; wherein, sodium alginate, water-soluble chitosan, and deionized water The mass ratio is 1:1:80;

[0029] (2) Calcium chloride and deionized water are mixed and stirred to a solid solution to obtain an aqueous calcium chloride solution; wherein the mass concentration of the aqueous calcium chloride solution is 5%;

[0030] (3) Place the mixed solution prepared above in a three-necked flask, then slowly dropwise add the prepared calcium chloride aqueous solution into the three-necked flask, stir while adding dropwise, and continue stirring for 30 minutes after the dropwise addition to obtain a mixed solution. Hydrogel; wherein, the mass ratio of the calcium chloride aqueous solution to the mixed ...

Embodiment 2

[0035] A preparation method of immobilized cellulase using modified nano-hydroxyapatite as a carrier, comprising the following steps:

[0036] (1) Mix sodium alginate, water-soluble chitosan and deionized water, and stir at 30°C±5°C until the solid dissolves to obtain a mixed solution; wherein, sodium alginate, water-soluble chitosan, and deionized water The mass ratio is 1:1:80;

[0037] (2) Calcium chloride and deionized water are mixed and stirred to a solid solution to obtain an aqueous calcium chloride solution; wherein, the mass concentration of the aqueous calcium chloride solution is 8%;

[0038] (3) Place the mixed solution prepared above in a three-necked flask, then slowly drop the prepared calcium chloride aqueous solution into the three-necked flask, stir while adding, after the dropwise addition, continue to stir for 30 minutes to obtain a mixed Hydrogel; wherein, the mass ratio of the calcium chloride aqueous solution to the mixed solution is 5:45; the dropping...

Embodiment 3

[0043] A preparation method of immobilized cellulase using modified nano-hydroxyapatite as a carrier, comprising the following steps:

[0044] (1) Mix sodium alginate, water-soluble chitosan and deionized water, and stir at 30°C±5°C until the solid dissolves to obtain a mixed solution; wherein, sodium alginate, water-soluble chitosan, and deionized water The mass ratio is 1:1:80;

[0045] (2) Calcium chloride and deionized water are mixed and stirred to a solid solution to obtain an aqueous calcium chloride solution; wherein, the mass concentration of the aqueous calcium chloride solution is 6%;

[0046] (3) Place the mixed solution prepared above in a three-necked flask, then slowly dropwise add the prepared calcium chloride aqueous solution into the three-necked flask, stir while adding dropwise, and continue stirring for 30 minutes after the dropwise addition to obtain a mixed solution. Hydrogel; wherein, the mass ratio of the calcium chloride aqueous solution to the mixed...

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PUM

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Abstract

The invention discloses a preparation method of modified nano-hydroxyapatite carried immobilized cellulase, and relates to the field of preparation of bio-enzyme preparations. Specifically, the method comprises the following steps: firstly, preparing modified nano-hydroxyapatite; then, preparing a cellulase water solution which is 1-2% in mass concentration, mixing and stirring a buffer solution which is constituted by the cellulase water solution as well as salicylic acid and sodium salicylate, regulating pH value of the solution to 3-5, adding the prepared modified nano-hydroxyapatite, and implementing stirring for 1-4h; and finally, adding a glutaraldehyde water solution which is 5% in mass concentration, conducting stirring at room temperature for 1-2h, then implementing filtering, washing an obtained solid by virtue of de-ionized water for 2-3 times, and implementing drying, so that the immobilized cellulase is obtained. The immobilized cellulase prepared by the invention is good in stability, excellent in reuse performance, safe and non-toxic, and low in preparation cost.

Description

Technical field: [0001] The invention relates to the field of preparation of biological enzyme preparations, in particular to a preparation method of immobilized cellulase using modified nano-hydroxyapatite as a carrier. Background technique: [0002] Cellulose is the most abundant renewable resource on the earth. Cellulase is a general term for a group of enzymes that degrade cellulose. It is mainly used in medicine, textiles, daily chemicals, papermaking, food fermentation, industrial washing, tobacco, and oil extraction. , wastewater treatment and feed and other fields. Cellulase and its substrates are very complex, with many influencing factors, slow hydrolysis rate, low enzymatic hydrolysis efficiency, and low enzyme utilization rate, so there are still many difficulties in practical application. [0003] Enzyme immobilization technology is a core technology in modern biotechnology and its industrialization. Immobilized enzymes have a wide range of applications in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08C12N11/04
CPCC12N9/2437C12N11/04C12N11/08C12N11/14C12Y302/01004
Inventor 冯海燕
Owner 苏州凯邦生物技术有限公司
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