Acetylcholinesterase chemiluminescence bioreactor, and preparation method and application thereof

The technology of cholinesterase chemistry and acetylcholinesterase is applied in the field of acetylcholinesterase chemiluminescence bioreactor and its preparation, and can solve the problems of the influence of the enzyme activity recovery rate, the lack of crucian carp brains, poisoning and the like, and achieves high recovery rate, The effect of short analysis time and good stability

Inactive Publication Date: 2011-02-09
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
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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...

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  • Acetylcholinesterase chemiluminescence bioreactor, and preparation method and application thereof
  • Acetylcholinesterase chemiluminescence bioreactor, and preparation method and application thereof
  • Acetylcholinesterase chemiluminescence bioreactor, and preparation method and application thereof

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Embodiment Construction

[0045] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0046] 1. Acetylcholinesterase chemiluminescent biosensor (referred to as sensor)

[0047] 1. Overall

[0048] Such as figure 1 , the sensor includes the 1st, 2nd, 3rd three-way valves 1, 2, 3, the reactor 4, the 1st, 2nd constant flow pumps 5, 6, the 1st, 2Y-type three-way 7, 8, the spiral tube 9, the dark box 10, photomultiplier tube 11, computer 12 and outlet 13 of waste liquid;

[0049] Also includes phosphate buffer solution a, ATCh substrate solution b, 2-PAM recovery agent c, sample d, potassium ferricyanide solution e and luminol solution f;

[0050] Phosphate buffer solution a and ATCh substrate solution b are communicated with the first three-way valve 1 respectively;

[0051] 2-PAM recovery agent c and sample d are communicated with the second three-way valve 2 respectively;

[0052] The first three-way valve 1 and the second three-way valve 2 communicate with the third t...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/46G01N21/76
Inventor 刘晓宇江丰赵冬冬石旺荣赵静邱瑾
Owner HUAZHONG AGRI UNIV
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