Method for extracting acetylcholinesterase by use of discarded organ-chicken heads in chicken manufacturing process
A technology of acetylcholinesterase and processing, which is applied in the direction of biochemical equipment and methods, enzymes, hydrolytic enzymes, etc., can solve the problems of high price, waste of grain raw materials, and complicated preparation process, and achieve easy storage, low cost, and high realization. The effect of value
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Embodiment 1
[0040] (1) Weigh the cleaned and dried watch glass, rinse the fresh longitudinally cut chicken head with physiological saline, take out the chicken brain with tweezers and put it in the watch glass, remove the non-brain tissue and weigh again, and calculate the chicken head. Brain tissue mass m;
[0041] (2) Mix chicken brain with phosphate buffer (0.1M, pH8.0) containing 1.0% (v / v) TritonX-100 in a ratio of 1:3 (g / mL);
[0042] (3) After homogenizing the mixed solution obtained in the previous step for 2 minutes with a homogenizer, let it stand for 30 minutes;
[0043] (4) Centrifuge the homogenate at 8000 r / min at 4°C for 20 min, and take the supernatant to obtain the crude extract of acetylcholinesterase. Place the obtained crude extract in a watch glass, and carry out vacuum freeze-drying to obtain an acetylcholinesterase product. Take 1.0 mg of the acetylcholinesterase product and dissolve it in distilled water to obtain an enzyme solution;
[0044] (5) Add 5mL of phosp...
Embodiment 2
[0048] (1) Weigh the cleaned and dried watch glass, rinse the fresh longitudinally cut chicken head with physiological saline, take out the chicken brain with tweezers and put it in the watch glass, remove the non-brain tissue and weigh again, and calculate the chicken head. Brain tissue mass m;
[0049] (2) Mix chicken brain with phosphate buffer (0.1M, pH8.0) containing 1.0% (v / v) TritonX-100 in a ratio of 1:4 (g / mL);
[0050] (3) After homogenizing the mixed solution obtained in the previous step for 2 minutes with a homogenizer, let it stand for 30 minutes;
[0051] (4) The homogenate was centrifuged at 8000r / min at 4°C for 20min, and the supernatant was collected to obtain the crude extract of acetylcholinesterase. Place the obtained crude extract in a watch glass, and carry out vacuum freeze-drying to obtain an acetylcholinesterase product. Take 1.0 mg of the acetylcholinesterase product and dissolve it in distilled water to obtain an enzyme solution;
[0052] (5) Add ...
Embodiment 3
[0056] (1) Weigh the cleaned and dried watch glass, rinse the fresh longitudinally cut chicken head with physiological saline, take out the chicken brain with tweezers and put it in the watch glass, remove the non-brain tissue and weigh again, and calculate the chicken head. Brain tissue mass m;
[0057] (2) Mix chicken brain with phosphate buffer (0.1M, pH8.0) containing 0.5% (v / v) TritonX-100 in a ratio of 1:4 (g / mL);
[0058] (3) After homogenizing the mixed solution obtained in the previous step for 2 minutes with a homogenizer, let it stand for 30 minutes;
[0059] (4) Centrifuge the homogenate at 8000 r / min at 4°C for 20 min, and take the supernatant to obtain the crude extract of acetylcholinesterase. Place the obtained crude extract in a watch glass, and carry out vacuum freeze-drying to obtain an acetylcholinesterase product. Take 1.0 mg of the acetylcholinesterase product and dissolve it in distilled water to obtain an enzyme solution;
[0060] (5) Put the supernat...
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