Acetylcholinesterase chemiluminescence bioreactor, and preparation method and application thereof

Abstract

The invention discloses an acetylcholinesterase chemiluminescence bioreactor, and a preparation method and application thereof, and belongs to the technical fields of analysis of pesticide residues and biotechnology. The reactor is a glass tube of which the two ends are sealed by 300-mesh bolting silk, wherein immobilized acetylcholinesterase is held in the glass tube; and the immobilized acetylcholinesterase is purified crucian muscle acetylcholinesterase which is immobilized on cyanogen bromide-activated (CNBr) sepharose 4B. The application of the reactor is characterized by comprising the following steps of: establishing the acetylcholinesterase chemiluminescence bioreactor; and detecting organic phosphorus and carbamate type pesticides. The crucian muscle acetylcholinesterase (AChE) extracted in the invention has high enzyme activity, obvious purification effect and high immobilized enzyme activity recovery rate; the detection limit of the organic phosphorus type pesticides and the carbamate type pesticides is low; the analysis time is short; and the enzyme reactor can be repeatedly used.

Description

technical field [0001] The invention belongs to the field of pesticide residue analysis and the field of biotechnology, and in particular relates to an acetylcholinesterase chemiluminescent bioreactor and its preparation method and application. Background technique [0002] Organophosphorus pesticides are a class of insecticides that were first synthesized by humans and are still widely used today. They are one of the most important pesticides currently used in my country. Organophosphorus pesticides have been widely used in agricultural production in my country, accounting for 77.76% of the total pesticide usage, due to their advantages such as high efficacy, many varieties, many control targets, and easy degradation in the environment. Since the 1970s, organochlorine pesticides have been banned or restricted by different countries, and the number of insects resistant to organophosphorus pesticides has increased, and the use of carbamate pesticides has increased year by yea...

AI Technical Summary

Problems solved by technology

But this patent has the following shortcomings: the patent uses crucian carp brain as the enzyme source, but the amount of crucian carp brain is too small, and the amount after purification by SephadexG-200 is even less

The patent adopts glutaraldehyde covalent cross-linking method to immobilize AChE. Although the binding degree is relatively firm and the binding reaction between the enzyme and the carrier can be completed quickly, glutaraldehyde itself has a toxic effect on AChE and will affect the recovery rate of enzyme activity. great influence

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