Purification method of acetylcholinesterase from earthworm serum
An acetylcholinesterase and earthworm technology, applied in the directions of hydrolase and the like, can solve the problems of not realizing the separation of esterase, low purification multiple, weakening enzyme activity, etc., and achieve the advantages of cheap and extensive enzyme sources, simple and stable preparation process, and low production cost. Effect
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[0014] see figure 1 , a method for purifying acetylcholinesterase from earthworm serum:
[0015] 1) Earthworm pretreatment: wash the purchased earthworms and put them in clean water for 24 hours to make them discharge the feces in the body, and store them in a refrigerator at 4°C for later use;
[0016] 2) Crude extraction of earthworm serum: use phosphate buffer (pH8.0, 0.1mol / L) as the solution to prepare the crude extract, and the homogenate ratio is 1:1 (weight of earthworm (g) / volume of phosphate buffer ( mL)), centrifuge at 4°C (10000r / min) for 20min, discard the supernatant, and obtain the crude enzyme extract, which is stored in the refrigerator. ;
[0017] 3) Cloud point extraction process: Take a 10mL centrifuge tube, then add 100μL of crude earthworm serum extract, that is, crude enzyme extract, then add surfactant Triton X-114 (0.09%, w / v) solution and 0.6mL pH4.1 citric acid buffer solution to adjust the pH value of the solution, dilute to 10mL with secondary w...
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