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Lactase film preparation method

A technology of lactase and mixed enzymes, which is applied in the field of biosensors and can solve problems such as not having universal applicability

Inactive Publication Date: 2010-12-08
TIANJIN UNIV OF COMMERCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the above method, the enzyme is directly immobilized on the electrode, and matching electrodes must be used, so it is not universally applicable

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Weigh 20g of β-cyclodextrin, add it to 100ml of 0.2M sodium hydroxide solution containing 1% glutaraldehyde by volume, heat to dissolve and stir for 20 minutes, leave it at room temperature for 12 hours, carry out cross-linking reaction, and then wash and precipitate with distilled water until the pH value of the washing liquid drops to about 7.0, and then fully grind it after drying in the dark, and the diameter of the milled particles is about 1nm, and then the aldylated β-cyclodextrin microspheres are prepared. Store in a 1.5 ml centrifuge tube protected from light.

[0017] Take 0.2 g of the immobilized material and 20 microliters of mixed enzyme solution and mix thoroughly, place it at 4 degrees Celsius for 8 minutes, take 0.1 g of the above-mentioned immobilized enzyme and spread it evenly on two polycarbonate microporous membranes, and then drop a drop of 2 % sodium alginate solution, after making the enzyme membrane of the sandwich structure, immerse the CaCl co...

Embodiment 2

[0020] Weigh 20g of β-cyclodextrin, add it to 100ml of 0.2M sodium hydroxide solution containing 1.5% glutaraldehyde by volume, heat to dissolve and stir for 20 minutes, leave it at room temperature for 12 hours, carry out cross-linking reaction, and then wash and precipitate with distilled water until the pH value of the washing liquid drops to about 7.0, and then fully grind it after drying in the dark, and the diameter of the milled particles is about 1nm, and then the aldylated β-cyclodextrin microspheres are prepared. Store in a 1.5 ml centrifuge tube protected from light.

[0021] Take 0.2 g of the immobilized material and 20 microliters of mixed enzyme solution and mix thoroughly, place it at 4 degrees Celsius for 8 minutes, take 0.1 g of the above-mentioned immobilized enzyme and spread it evenly on two polycarbonate microporous membranes, and then drop a drop of 2 % sodium alginate solution, after making the enzyme membrane of the sandwich structure, immerse in CaCl w...

Embodiment 3

[0024] Weigh 20g of β-cyclodextrin, add it to 100ml of 0.2M sodium hydroxide solution containing 0.5% glutaraldehyde by volume, heat to dissolve and stir for 20 minutes, leave it at room temperature for 12 hours, carry out cross-linking reaction, and then wash and precipitate with distilled water until the pH value of the washing liquid drops to about 7.0, and then fully grind it after drying in the dark, and the diameter of the milled particles is about 1nm, and then the aldylated β-cyclodextrin microspheres are prepared. Store in a 1.5 ml centrifuge tube protected from light.

[0025] Take 0.2 g of the immobilized material and 20 microliters of mixed enzyme solution and mix thoroughly, place it at 4 degrees Celsius for 8 minutes, take 0.1 g of the above-mentioned immobilized enzyme and spread it evenly on two polycarbonate microporous membranes, and then drop a drop of 2 % sodium alginate solution, after making the enzyme film of sandwich structure, immerse in CaCl with a co...

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Abstract

The invention discloses a preparation method of lactase membrane, and aims to provide a core component - nucleopore immobilized enzyme membrane which can be matched with different redox electrodes, and is fit for lactase online monitoring. The method includes the following steps: beta-cyclodextrin is weighed and dissolved in low concentration sodium hydroxide solution with glutaric dialdehyde, and is placed in room temperature, so that the two are crosslinked into solid state, a precipitated crystal is washed with distilled water, and milled after being dried away from light, aldehyde group beta-cyclodextrin microsphere is prepared; the immobilized material is taken and sufficiently mixed with hybrid enzyme solution, and placed for 8 minutes at 4 degrees centigrade; immobilized enzyme is taken and uniformly applied on two polycarbonate microporous membranes, and dripped with sodium alginate solution; and an enzyme membrane with a sandwich structure is prepared, taken out after being dipped into CaCl2 solution, and washed by buffer solution to be prepared into lactase membrane. The nucleopore immobilized enzyme membrane prepared by the method of the invention can be flexibly matched with other redox electrodes, and has general applicability.

Description

technical field [0001] The invention relates to the field of biosensors, in particular to a method for preparing a lactase membrane. Background technique [0002] The determination methods of lactose content mainly include: reducing sugar determination method (iodometric method), polarimetry, near-infrared spectroscopy, nuclear magnetic resonance method, and high performance liquid chromatography, ultrasonic method, etc. These methods are time-consuming, cumbersome, and difficult to detect It is time-consuming and labor-intensive. Therefore, it is generally optimistic about the enzyme assay method, namely lactose biosensor method, which has the characteristics of sensitivity, rapidity, simplicity, and strong specificity, and hardly requires any pretreatment of samples. [0003] The development of the lactose sensor provides a favorable means for the on-site rapid detection of lactose. A variety of biosensors have emerged, among which the amperometric enzyme sensor is still...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
Inventor 庞广昌李家鹏
Owner TIANJIN UNIV OF COMMERCE