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Bionic nanometer grain preparation method of sio2 fixed beta-glucuronide enzyme

A technology of aldolidase and silicon dioxide, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of activity decline and inactivation, achieve good immobilization ability, mild preparation conditions, and good stability for repeated use Effect

Inactive Publication Date: 2010-06-09
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is simple and easy to implement, the alcohol by-products produced during the process have an adverse effect on the maintenance of the activity of the biological enzyme, resulting in a decrease in activity or even complete inactivation.

Method used

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  • Bionic nanometer grain preparation method of sio2 fixed beta-glucuronide enzyme
  • Bionic nanometer grain preparation method of sio2 fixed beta-glucuronide enzyme
  • Bionic nanometer grain preparation method of sio2 fixed beta-glucuronide enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Accurately weigh 1.0mg of β-glucuronidase, dissolve it in 0.05mol / L tris-hydrochloric acid (Tris-HCl) buffer solution, and dilute to 10mL to obtain 0.1mg / mL β-glucuronidase Acidase solution. Weigh 300 mg of protamine sulfate, dissolve it in 0.05 mol / L Tris-HCl buffer solution, and dilute to 20 mL to obtain a 15 mg / mL protamine sulfate solution. Sodium silicate was dissolved in deionized water, and the pH of the solution was adjusted to 7.0 with 2mol / L HCl to prepare a 10mmol / L sodium silicate solution.

[0017] Take 0.5ml each of β-glucuronidase solution and protamine sulfate solution, mix well and add to 7mL freshly prepared sodium silicate solution, let it stand for 5min, set the centrifuge speed to 4500r / min, and centrifuge for 10min . Pour out the supernatant, wash with deionized water, centrifuge for 10 min, and pour out the supernatant. Wash repeatedly 2-3 times. The content of β-glucuronidase in the supernatant was analyzed through the conversion reaction of ...

Embodiment 2

[0021] Accurately weigh 1.0mg of β-glucuronidase, dissolve it in 0.05mol / L tris-hydrochloric acid (Tris-HCl) buffer solution, and dilute to 10mL to obtain 0.1mg / mL β-glucuronidase Acidase solution. Weigh 40 mg of protamine sulfate, dissolve it in 0.05 mol / L Tris-HCl buffer solution, and dilute to 20 mL to obtain a 2 mg / mL protamine sulfate solution. Sodium silicate was dissolved in deionized water, and the pH of the solution was adjusted to 7.0 with 2mol / L HCl to prepare a 30mmol / L sodium silicate solution.

[0022] Take 0.5mL of β-glucuronidase solution and protamine sulfate solution, mix well and add to 7mL freshly prepared sodium silicate solution, let it stand for 10min, set the centrifuge speed to 4500r / min, and centrifuge for 10min . Pour out the supernatant, wash with deionized water, centrifuge for 10 min, and pour out the supernatant. Wash repeatedly 2-3 times. The content of β-glucuronidase in the supernatant was analyzed through the conversion reaction of baical...

Embodiment 3

[0026] Accurately weigh 1.0 mg of β-glucuronidase, dissolve it in deionized water, and dilute to 10 ml to obtain 0.1 mg / ml β-glucuronidase solution. Weigh 300 mg of protamine sulfate, dissolve it in deionized water, and dilute to 20 ml to obtain a 15 mg / ml protamine sulfate solution. Sodium silicate was dissolved in deionized water, and the pH of the solution was adjusted to 7.0 with 2mol / L HCl to prepare a 0.03mmol / L sodium silicate solution.

[0027] Take 0.5mL of β-glucuronidase solution and protamine sulfate solution, mix well and add to 10mL freshly prepared sodium silicate solution, let it stand for 5min, set the centrifuge speed to 4500r / min, and centrifuge for 10min . Pour out the supernatant, wash with deionized water, centrifuge for 10 min, and pour out the supernatant. Wash repeatedly 2-3 times. The content of β-glucuronidase in the supernatant was analyzed through the conversion reaction of baicalin, and it was found that there was no β-glucuronidase in the supe...

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Abstract

The invention discloses a preparation method of Beta- glucuronidase nanometer particle which is immobilized by bionic silicon dioxide. The process of the method comprises that the protamine sulfate isdissolved in the trishydroxymethylaminomethane-hydrochloride buffer or the deionized water for the preparation of protamine sulfate solution; the Beta- glucuronidase is dissolved in the trishydroxymethylaminomethane-hydrochloride buffer or the deionized water for the preparation of Beta- glucuronidase solution; the protamine sulfate solution is mixed with the Beta- glucuronidase solution to obtain the mixed solution; the sodium silicate is dissolved in the deionized water or the saline water and the pH value is adjusted to obtain the sodium silicate solution; the mixed solution is added intothe sodium silicate solution; through standing, separation, removal of the supernatant, washing and freeze drying, the Beta- glucuronidase nanometer particle which is immobilized by bionic silicon dioxide is obtained. The invention has the advantages that the preparation process is simple; the immobilized Beta- glucuronidase has a high activity maintenance rate and good stability for repeat use.

Description

technical field [0001] The invention relates to a method for preparing nanoparticles of biomimetic silicon dioxide immobilized β-glucuronidase, which belongs to the enzyme immobilization technology. Background technique [0002] Silica gel is a commonly used carrier for immobilized enzymes. At present, silica is used as a carrier to immobilize enzymes mainly by embedding or adsorption. The adsorption method is simple to operate, but the enzyme is easy to fall off, and the reusability is poor. The high loading rate of the embedding method is conducive to maintaining the structural integrity of the enzyme, thereby improving the activity maintenance rate of the enzyme, and the silica has a mesoporous structure, which is conducive to the transfer of reactants and products. [0003] At present, the sol-gel (sol-gel) method is generally used, using acid or alkali as a catalyst, using the hydrolysis and polycondensation of sodium silicate or organosilane precursors to synthesize ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14
Inventor 吴洪姜忠义李林张羽飞
Owner TIANJIN UNIV
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