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

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...