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Method for preparing immobilized enzyme based on reaction adsorption method

An enzyme immobilization and adsorption technology, applied in the direction of being immobilized on or in an inorganic carrier, immobilized on or in an organic carrier, etc., can solve the problems of weak adsorption force, easy free shedding of the enzyme, etc. Stable, easy to operate, and quick to complete

Active Publication Date: 2021-04-02
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the main disadvantage of these adsorption methods is that the adsorption force is weak, causing the enzyme to dissociate easily. If the shortcomings of the adsorption method can be well overcome, considering the overall cost, it is still a very practical immobilization technology.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Immobilized esterase:

[0030] Cultivate the bacterial strain S.mal SF-H1 that produces esterase, ferment for 72 hours, the esterase activity of the fermentation broth is 21u / ml, filter and sterilize to obtain the fermented esterase crude enzyme liquid, take 200ml of the fermented esterase crude enzyme liquid and add 10ml of 1M chloride Calcium, at room temperature, pH 6.0, after stirring evenly, add 10ml of 1M dipotassium hydrogen phosphate (sodium) solution, stir quickly to form a white precipitate containing immobilized esterase, centrifuge, wash the precipitate three times with water, take a sample to determine the immobilized enzyme Activity, 61% of the total enzyme activity of one-time immobilization. The immobilized enzyme is used continuously. After each catalytic substrate hydrolysis is completed, the immobilized enzyme is centrifuged and washed three times for the next use. After 20 times, the enzyme activity 85% remained, indicating that the enzyme was firmly...

Embodiment 2

[0032] Immobilized protease:

[0033] Take solid powder protease and prepare 200mg / 200ml enzyme solution, filter to remove impurities, add 10ml of 1M calcium chloride, at room temperature, pH6.0, after stirring evenly, add 10ml of 1M dipotassium hydrogen phosphate (sodium) solution, stir quickly to form The precipitate containing immobilized enzyme was centrifuged, and the precipitate was washed three times with water. The immobilized protease was sampled and determined. The total enzyme activity of one-time immobilized was 67%. The immobilized enzyme was used continuously for 10 times, and the remaining enzyme activity was 82%. The immobilization is firm and not easy to fall off.

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PUM

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Abstract

The invention discloses a method for preparing immobilized enzyme based on a reactive adsorption method, and relates to the technical field of bionts. The method comprises the following steps: enabling reaction between two or more compounds in an enzyme-containing aqueous solution, and adsorbing and wrapping enzyme molecules by precipitates in a process of forming one or more precipitates such as calcium phosphate, calcium hydrogen phosphate, calcium carbonate, calcium hydrogen carbonate, aluminum hydroxide, calcium phytic acid, zinc ferrocyanide and chitosan organic matters to prepare the immobilized enzyme. The method is an improved adsorption method, aims to increase the bonding force between a carrier and the enzyme and to overcome the shortcoming that the immobilized enzyme has high probability of falling off, has the characteristics of simple operation, low cost and stable enzyme activity, and is worthy of being popularized and applied.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing immobilized enzymes based on a reaction adsorption method. Background technique [0002] Enzymes are an important class of organic macromolecules that exist in living organisms. As biocatalysts, they participate in many biochemical metabolic activities in the body. Compared with inorganic catalysts, enzymes have the characteristics of strong specificity, high catalytic efficiency and mild reaction conditions. Enzyme molecules isolated from plants, animals or microorganisms that catalyze specific reactions can also be used to catalyze specific reactions in vitro. These obvious advantages of enzymes have greatly promoted the production and application of enzymes. [0003] However, the source of the enzyme determines that the price of the enzyme is relatively expensive. After the reaction, it is difficult to conveniently separate, recover and reuse it from the wat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12N11/10
CPCC12N11/10C12N11/14
Inventor 王丽丽王崇惠赵一张春晓
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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