Butyrylcholine esterase, preparation method thereof and kit for quickly detecting pesticide residue and prepared by butyrylcholine esterase

Abstract

The invention discloses a butyrylcholine esterase, a preparation method thereof and a kit for quickly detecting pesticide residue and prepared by butyrylcholine esterase. The preparation method comprises the following steps: performing purification to bovine serum, homogenating, extracting via the buffer solution, and centrifuging to remove sediment so as to obtain crude enzyme; precipitating the crude enzyme via ammonium sulfate, centrifuging, removing the liquid supernatant, redissolving and dialyzing the sediment, performing chromatography via the DEAE column, collecting eluant, dialyzing, obtaining the enzyme after purification, and then freezing and drying; the kit comprises butyrylcholine esterase, substrate, developer and phosphoric acid buffer solution; the concentration of the phosphate buffer is 0.9-0.11 mol / L, and the pH value of the phosphate buffer is 7.5-8.5. The kit of the invention adopts the matching of semi-purified butyrylcholine esterase to be matched with the stabilizing agent, so that the stability of butyrylcholine esterase is remarkably enhanced; the butyrylcholine esterase is obtained through the purification of the bovine serum, the sensitivity is strong, and the sensitivity of the kit prepared by the butyrylcholine esterase is strong.

Description

technical field [0001] The invention relates to the field of detection reagents, in particular to butyrylcholinesterase, a preparation method thereof and a kit for rapidly detecting pesticide residues prepared therefrom. Background technique [0002] As an important crop in my country, sugarcane is not only the most important raw material for sugar production, but also can produce alcohol and brew edible wine. Sugarcane production has long relied on highly toxic and high-residue organic phosphorus and carbamate pesticides such as carbofuran, phorate, terbufos, and isofenphos-methyl. The extensive use and abuse of such pesticides has negative impacts on sugarcane production and ecology. The negative impact on the environment has become increasingly prominent, and food poisoning incidents caused by excessive pesticide residues have occurred from time to time. Although my country's "Sugar Hygienic Standard" GB13104-2005 clearly stipulates that pesticide residues in the product...

AI Technical Summary

Problems solved by technology

[0004] Enzymes commonly used in the detection of pesticide residues include carboxylesterase (mainly derived from animal liver, plants, etc.), butyrylcholinesterase and acetylcholinesterase. Acetylcholinesterase is the most commonly used substrate for organophosphorus and carbamate agents. However, acetylcholinesterase is mainly extracted from the heads of sensitive insects such as houseflies, with low yield and high cost, which is far from meeting the current requirements for residue detection; butyrylcholinesterase is extracted from animal serum with high yield. The cost is low, how to improve the sensitivity and stability of butyrylcholinesterase is very important

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