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Method for preparing saccharose biosensor nuclear micropore enzyme membrane using beta-galactosidase

A technology of biosensor and galactosidase, which is applied in the field of preparation of sucrose biosensor nuclear micropore enzyme membrane, can solve the problems of large error and complicated process, and achieve the effects of simplified production process, excellent sensitivity and stability

Inactive Publication Date: 2009-06-24
ENSOUL TECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been reports on the successful development of sucrose biosensors in foreign countries, but more than 3 kinds of enzymes are needed to detect, the error is large, and the process is complicated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Preparation of a mixed enzyme solution: 0.005 g of glucose oxidase, 1 mL of β-galactosidase with an activity unit of 3000 units / mL, and 0.01 g of bovine serum albumin were thoroughly mixed to prepare a mixed enzyme solution.

[0015] Weigh 0.7g β-cyclodextrin and dissolve it in 10m1 of a mixed solution consisting of NaOH with a concentration of 0.2Mo1 / L and 5% glutaraldehyde in a low-concentration sodium hydroxide solution. After that, 60°C Heat treatment until the crystals are dispersed and a cross-linking reaction occurs to form poly-β-cyclodextrin with active aldehyde groups. Then it is washed with distilled water to form poly-β-cyclodextrin with active aldehyde groups until the NaOH on the surface of poly-β-cyclodextrin is completely removed, dried and then ground into particles with a diameter of 0.8-1.2 mm.

[0016] 0.1 g of the above-mentioned polyβ-cyclodextrin particles with active aldehyde groups as immobilization material was fully mixed with 10 μl of the abo...

Embodiment 2

[0018] Preparation of a mixed enzyme solution: 0.007 g of glucose oxidase, 1 mL of β-galactosidase with an activity unit of 3000 units / mL, and 0.01 g of bovine serum albumin were thoroughly mixed to prepare a mixed enzyme solution.

[0019] Weigh 0.7g β-cyclodextrin and dissolve it in 10ml of a mixed solution consisting of NaOH with a concentration of 0.2Mol / L and 2.5% glutaraldehyde in a low-concentration sodium hydroxide solution. After that, 60°C Heat treatment until the crystals are dispersed and a cross-linking reaction occurs to form poly-β-cyclodextrin with active aldehyde groups. Then it is washed with distilled water to form poly-β-cyclodextrin with active aldehyde groups until the NaOH on the surface of poly-β-cyclodextrin is completely removed. After drying, it is ground into particles with a diameter of about 0.8-1.2 mm.

[0020] 0.1 g of the above-mentioned polyβ-cyclodextrin particles with active aldehyde groups as immobilization material was fully mixed with 10 ...

Embodiment 3

[0022] Preparation of a mixed enzyme solution: 0.003 g of glucose oxidase, 1 mL of β-galactosidase with an activity unit of 3000 units / mL, and 0.01 g of bovine serum albumin were thoroughly mixed to prepare a mixed enzyme solution.

[0023] Weigh 0.7g of β-cyclodextrin and dissolve it in 10ml of a mixed solution consisting of NaOH with a concentration of 0.2Mol / L and 3% glutaraldehyde in a low-concentration sodium hydroxide solution. Heat treatment until the crystals are dispersed and a cross-linking reaction occurs to form poly-β-cyclodextrin with active aldehyde groups. Then it is washed with distilled water to form poly-β-cyclodextrin with active aldehyde groups until the NaOH on the surface of poly-β-cyclodextrin is completely removed, dried and then ground into particles with a diameter of 0.8-1.2 mm.

[0024] 0.1 g of the above-mentioned polyβ-cyclodextrin particles with active aldehyde groups as immobilization material was fully mixed with 10 μl of the above-prepared mi...

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Abstract

The invention discloses a method for preparing a nuclear pore enzyme membrane of a sucrose biosensor by using beta-galactosidase, aiming at providing a method for obtaining the nuclear pore enzyme membrane of the sucrose biosensor only by immobilizing two enzymes. The method comprises the steps: beta-cyclodextrin is weighed and taken to be dissolved in the mixed solution formed by low concentration NaOH and glutaric dialdehyde; after that, the mixture is heated and treated at the temperature of 60 DEG C until the crystalloid is dispersed, and poly beta-cyclodextrin having active aldehyde group is formed by cross linking reaction; then, the poly beta-cyclodextrin having the active aldehyde group is washed by distilled water until the NaOH on the surface of the poly beta-cyclodextrin is thoroughly removed; after being dried at the room temperature, the poly beta-cyclodextrin is milled into granules basically having uniform size; the poly beta-cyclodextrin granules having the active aldehyde group are taken as fixing material and then is fully mixed with mixed enzyme solution to have reaction for 80 minutes at 25 DEG C to cause the enzyme immobilization; the immobilized enzyme is evenly coated on the surfaces of two nuclear pore membranes to be made into the nuclear pore enzyme membrane of the sucrose biosensor having a sandwich structure.

Description

technical field [0001] The invention relates to the field of biosensors, in particular to a method for preparing a sucrose biosensor nuclear micropore enzyme membrane. Background technique [0002] Sucrose is a non-reducing disaccharide and cannot be determined by conventional methods for the determination of reducing sugars. At present, in order to meet the needs of rapid and sensitive detection, the detection method of sucrose biosensor is adopted. There have been reports on the successful development of sucrose biosensors in foreign countries, but more than three enzymes are needed for detection, resulting in large errors and complicated processes. Contents of the invention [0003] The invention aims to overcome the shortcomings in the prior art, and provides a method for obtaining the nuclear micropore enzyme membrane of the sucrose biosensor by only immobilizing two kinds of enzymes. [0004] The present invention realizes through following technical scheme: [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12C12N11/08G01N27/327
Inventor 庞广昌李加鹏吕瑜峰陈庆森胡志和
Owner ENSOUL TECH LTD
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