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Silicon-based compound immobilized enzyme biocatalyst and preparation method thereof

A technology of silicon-based compounds and biocatalysts, applied to biochemical equipment and methods, enzymes immobilized on or in inorganic carriers, etc., can solve problems such as loss of enzyme activity

Active Publication Date: 2021-04-27
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its binding force is strong, the enzyme is not easy to fall off, and the stability is good, but its violent reaction is also easy to cause the loss of enzyme activity

Method used

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  • Silicon-based compound immobilized enzyme biocatalyst and preparation method thereof
  • Silicon-based compound immobilized enzyme biocatalyst and preparation method thereof
  • Silicon-based compound immobilized enzyme biocatalyst and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] See attached figure 1 Process flow chart in . First prepare the silicon-based compound carrier. Hollow glass beads N20 were used as raw materials, placed in 2M NaOH solution, refluxed and stirred at 100° C. for 3 hours to obtain pulverized silicon-based compounds. The powdered silicon-based compound is used as the core, and tetraethylorthosilicate (TEOS) is used as the source of encapsulated silicon. First weigh 0.5g of powdered silicon-based compound into a beaker, add 36mL of ethanol, 2mL of tetraethyl orthosilicate, 100μL of 0.5mol / L sulfuric acid, hydrolyze at 50°C for 6h, and concentrate at 80°C for 18h to form a white powder. Take 0.3g of powdered silicon-based compound, add 30g of toluene and 3g of vinyltriethoxysilane, and stir at reflux at 110°C for 3h. Wash with toluene and dichloromethane three times each. Such as Figure 4 As shown, the prepared carrier particle size is between 20-80 μm; as Figure 5 It can be seen that after the carrier is modified, a...

Embodiment 2

[0044] The preparation steps of the modified silicon-based compound in this example are the same as the preparation steps of the modified silicon-based compound in Example 1: weigh 0.5g of powdered silicon-based compound in a beaker, add 30g of ethanol, 2g of tetraethyl orthosilicate , 100μL of 0.5mol / L sulfuric acid, respectively placed at 30°C, 40°C, 50°C, 60°C, 70°C for 6h, and concentrated at 80°C for 18h to form a white powder. Take 0.3g of powdered silicon-based compound, add 30g of toluene and 3g of vinyltriethoxysilane, and stir under reflux at 110°C for 3h. Wash with toluene and dichloromethane three times each. After blowing dry with nitrogen gas, 10 mL of lipase solution was added, stirred and fixed at 30° C. for 3 h to obtain an immobilized enzyme biocatalyst. attached by Figure 8 It can be seen that the enzyme activity recovery rate of immobilized enzyme activity increases with the increase of temperature. When the temperature is 60°C, the enzyme activity recov...

Embodiment 3

[0046] The preparation steps of the modified silicon-based compound in this example are the same as the preparation steps of the modified silicon-based compound in Example 1: weigh 0.5g powdered silicon-based compound in a beaker, 30g ethanol, 2g tetraethyl orthosilicate, Add 100 μL of sulfuric acid with a concentration of 0.01mol / L, 0.05mol / L, 0.1mol / L and 0.5mol / L respectively, hydrolyze at 50°C for 6 hours, and concentrate at 80°C for 18 hours to form a white powder. Take 0.3g of powdered silicon-based compound, add 30g of toluene and 3g of vinyltriethoxysilane, and stir under reflux at 110°C for 3h. Wash with toluene and dichloromethane three times each. After blowing dry with nitrogen gas, 10 mL of lipase solution was added, stirred and fixed at 30° C. for 3 h to obtain an immobilized enzyme biocatalyst. attached by Figure 9 It can be seen that the recovery rate of the enzyme activity of the immobilized enzyme activity increases with the increase of the acid concentrat...

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Abstract

The invention provides a preparation method of a silicon-based compound immobilized enzyme biocatalyst, and belongs to the technical field of preparation of immobilized enzyme biocatalysts. According to the method, a silicon-based compound is used as a carrier, lipase is used as an immobilization object, a covalent binding method is used as an immobilization method, and the surface of the silicon-based compound is rich in hydroxyl groups and is covalently bound with a cross-linking agent rich in ethoxy groups. The other end of the cross-linking agent can be covalently combined with an active group on the lipase to achieve the effect of the immobilized enzyme. The immobilized enzyme catalyst prepared by the method disclosed by the invention has good dispersibility, high enzyme loading capacity and good enzyme activity. The method is also suitable for immobilization of laccase, horse radish peroxidase and other enzymes.

Description

technical field [0001] The invention belongs to the field of preparation of immobilized enzyme biocatalysts, in particular to a silicon-based compound immobilized enzyme biocatalyst and a preparation method thereof. Background technique [0002] Immobilized enzyme (Immobilized Enzyme) is a concept proposed in the first International Enzyme Engineering Conference in 1917, that is, by physical or chemical methods, the enzyme is bound on a water-insoluble carrier or in a certain space, and the free movement of the free enzyme is restricted. And it can make the enzyme play a catalytic role, and the enzyme can be easily separated from the substrate and product after the reaction, so as to achieve the purpose of repeated use. After a century of development, the immobilized enzymes are mainly divided into four methods: 1) Adsorption method: The adsorption method is to immobilize the enzyme on a specific carrier through non-specific physical adsorption or specific adsorption between...

Claims

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Application Information

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IPC IPC(8): C12N11/14
CPCC12N11/14C12N9/20C12Y301/01003
Inventor 石贵阳李由然李涛卢佳伟张梁丁重阳徐沙顾正华
Owner JIANGNAN UNIV
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