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Method for preparing immobilized alpha-amylase by using spherical tannin as carrier

An amylase and tannin technology is applied in the field of preparation of immobilized alpha-amylase to achieve the effects of uniform shape, large surface area and improved affinity

Inactive Publication Date: 2010-12-15
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on immobilizing α-amylase by adsorption-crosslinking method using the spherical tannin as the immobilized enzyme carrier

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] The "preparation method of a spherical tannin carrier" is used to firstly prepare a spherical tannin carrier, and use the prepared spherical tannin carrier to immobilize α-amylase.

[0010] Take 0.5g of spherical tannin carrier and add it to 50ml of α-amylase solution with a content of 2.5mg / ml, adsorb at 30°C for 5 hours, add 2ml of glutaraldehyde solution with a concentration of 20%, and put it at 30°C After cross-linking under temperature conditions for 2 hours, the filtrate was removed by filtration, washed with a phosphate buffer solution with a pH value of 7 and blotted dry to obtain spherical tannin-immobilized α-amylase.

[0011] It was determined that the recovery rate of the enzyme activity reached 82%; compared with the solution enzyme, the affinity of the immobilized α-amylase to the substrate (starch) was increased by 14.3 times.

Embodiment 2

[0013] The method for preparing the spherical tannin carrier is the same as in Example 1.

[0014] The spherical tannin carrier was prepared first by using "a preparation method of a spherical tannin carrier", and the α-amylase was immobilized with the prepared spherical tannin carrier.

[0015] Take 0.5g of spherical tannin carrier and add it to 100ml of α-amylase solution with a content of 9.5mg / ml, adsorb at 30°C for 5 hours, add 5ml of glutaraldehyde solution with a concentration of 20%, and put it at 30°C After cross-linking under certain conditions for 2 hours, the filtrate was removed by filtration, washed with a phosphate buffer solution with a pH value of 7, and blotted dry to obtain spherical tannin-immobilized α-amylase.

[0016] It was determined that the recovery rate of enzyme activity reached 81.8%; compared with solution enzyme, the affinity of immobilized α-amylase to substrate (starch) was increased by 13.2 times.

Embodiment 3

[0018] The method for preparing the spherical tannin carrier is the same as in Example 1.

[0019] Take 0.5g of spherical tannin carrier and add 100ml of 3mg / ml to the α-amylase solution, absorb at 30°C for 5 hours, add 5ml of glutaraldehyde solution with a concentration of 25%, and put it at 30°C After cross-linking for 2 hours, the filtrate was removed by filtration, washed with a phosphate buffer solution with a pH value of 7 and blotted dry to obtain a spherical tannin-immobilized α-amylase.

[0020] It was determined that the recovery rate of enzyme activity reached 85.3%. Compared with solution enzymes, the affinity of immobilized α-amylase to substrate (starch) was increased by 15.9 times.

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Abstract

The invention relates to a method for preparing immobilized alpha-amylase, in particular to a method for preparing immobilized alpha-amylase by using spherical tannin as a carrier. The method is characterized by comprising the following steps: 1) adding the spherical tannin into solution of alpha-amylase to obtain mixed solution of the spherical tannin carrier and the alpha-amylase; 2) performing an absorption action in the mixed solution at 4 to 50 DEG C, adding glutaraldehyde solution, and performing crosslinking at 4 to 50 DEG C; and 3) filtering to remove filtrate, washing with neutral phosphate buffer solution and drying by absorption to obtain the alpha-amylase immobilized on the spherical tannin. The prepared alpha-amylase immobilized on the spherical tannin has uniform shape and large surface area; the average amylase activity recovery rate is up to over 80 percent; and compared with solution enzyme, the affinity of the immobilized alpha-amylase with a substrate (starch) is improved by more than 12 times.

Description

technical field [0001] The invention relates to a preparation method of immobilized alpha-amylase, in particular to a preparation method of immobilized alpha-amylase with globular tannin as carrier. Background technique [0002] α-amylase (α-amylase) cuts the α-1 and 4 bonds from the inside of the starch molecule, and decomposes the starch into dextrin and reducing sugar. It is widely used in grain and food industry, brewing, fermentation, textile industry and pharmaceutical industry. However, since the enzyme is a soluble protein, it is not easy to recycle and reuse, resulting in a waste of catalyst, and the enzyme protein will also pollute the product, causing difficulties in post-processing. [0003] Enzyme immobilization technology is a new type of biotechnology developed in the 1960s, which is defined as "an enzyme that is bound in a certain area by chemical or physical means, maintains catalytic activity, and can be used repeatedly and continuously." The use of immobi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02
Inventor 卢玉栋吴宗华游瑞云
Owner FUJIAN NORMAL UNIV
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