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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

Inactive Publication Date: 2015-09-02
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on plant esterase has not been in-depth. Some studies have shown that the enzyme activity in food crops is relatively high, but for most food raw materials, the extraction of acetylcholinesterase will also cause waste of food raw materials
Due to the existence of problems such as enzyme sources and complex preparation processes, the price is expensive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention provides a method for extracting acetylcholinesterase by use of discarded organ-chicken heads in the chicken manufacturing process. The discarded organ-chicken brain during chicken processing is used as the material source of enzyme. The method comprises the following steps: processing of chicken brains, preparation and refrigerated centrifuge of a homogenate and vacuum freeze drying of a crude extract, and through the steps, the acetylcholinesterase product can be obtained. As the acetylcholinesterase product prepared from traditional enzyme sources is high in cost and low in yield while the imported enzyme sources are high in price and relatively long in cycle, through screening of enzyme sources, the discarded organs-chicken brains in the chicken manufacturing process are fully utilized to achieve high-value utilization and reduce the cost, and through optimization and inspection, the enzyme activity is relatively high, the enzyme activity of the crude enzyme can reach 3.0-5.0 U / mL, the specific activity can reach 0.35-0.5 U / mg, and accordingly, a foundation is laid for follow-up preparation of the pure acetylcholinesterase product.

Description

technical field [0001] The invention relates to an enzyme extraction method, in particular to a method for extracting acetylcholinesterase by using the discarded organ-chicken head in the chicken processing process. Background technique [0002] Acetylcholinesterase (AChE) is a vital enzyme in the process of cholinergic nerve transmission, it can rapidly hydrolyze the neurotransmitter acetylcholine (ACh) into acetic acid and choline, thereby terminating the action of ACh on the postsynaptic The excitatory effect of the membrane ensures the normal transmission of nerve signals in the organism. Due to the high-speed hydrolysis of AChE to ACh, it has attracted more and more researchers' attention in recent years. AChE has a wide range of applications in biological control, pesticide residue detection, molecular biology and treatment of various neurological diseases. Recent studies have shown that ethyl carbamate can inhibit the activity of AChE to a certain extent, so AChE ma...

Claims

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Application Information

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IPC IPC(8): C12N9/18
CPCC12N9/18C12Y301/01007
Inventor 黄惠华张佳佳黄秋婷
Owner SOUTH CHINA UNIV OF TECH
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