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
2 Cites 1 Cited by

AI-Extracted 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 extracti...
View more

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.

Application Domain

Hydrolases

Technology Topic

ChemistryFreeze dry +8

Examples

  • Experimental program(3)

Example Embodiment

[0039] Example 1:
[0040] (1) Weigh the cleaned and dried watch glass, rinse the fresh chicken heads cut vertically with normal saline, use tweezers to take the chicken brain out of the watch glass, remove the non-brain tissue, and weigh it again to calculate the chicken Brain tissue quality m;
[0041] (2) Mix the chicken brain with phosphate buffer (0.1M, pH8.0) containing 1.0% (v/v) TritonX-100 at a ratio of 1:3 (g/mL);
[0042] (3) After homogenizing the mixture obtained in the previous step with a homogenizer for 2 minutes, let it stand for 30 minutes;
[0043] (4) Centrifuge the homogenate at 8000r/min at 4°C for 20min, and take the supernatant to obtain the crude acetylcholinesterase extract. Place the obtained crude extract in a watch glass and perform vacuum freeze-drying to obtain an acetylcholinesterase product. Take 1.0 mg of the acetylcholinesterase product and dissolve it with distilled water to obtain an enzyme solution;
[0044] (5) In a 10mL colorimetric tube, add 5mL phosphate buffer (0.1M, pH7.5), then add 0.02mL enzyme solution to mix well, and keep it in a constant temperature water bath at 36.5℃ for 15min;
[0045] (6) Add 0.1mL thioacetylcholine iodide solution (20mg/mL) and 0.1mL DTNB (10mg/mL) solution to the solution obtained in the previous step and mix well;
[0046] (7) Use a cuvette in the 412nm wavelength range to compare the color, record the absorbance A = 0.9876, measure the total protein content by the Coomassie brilliant blue method, and then calculate it according to the formula of enzyme activity, specific activity and total enzyme activity, and the enzyme activity is 6.3178U/mL, the specific activity is 0.4254U/mg.

Example Embodiment

[0047] Example 2:
[0048] (1) Weigh the cleaned and dried watch glass, rinse the fresh chicken heads cut vertically with normal saline, use tweezers to take the chicken brain out of the watch glass, remove the non-brain tissue, and weigh it again to calculate the chicken Brain tissue quality m;
[0049] (2) Mix the 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 mixture obtained in the previous step with a homogenizer for 2 minutes, let it stand for 30 minutes;
[0051] (4) Centrifuge the homogenate at 8000r/min at 4°C for 20min, and take the supernatant to obtain the crude acetylcholinesterase extract. Place the obtained crude extract in a watch glass and perform vacuum freeze-drying to obtain an acetylcholinesterase product. Take 1.0 mg of the acetylcholinesterase product and dissolve it with distilled water to obtain an enzyme solution;
[0052] (5) In a 10 mL colorimetric tube, add 5 mL of phosphate buffer (0.1M, pH 7.5), then add 0.02 mL of enzyme solution to mix, and keep it in a constant temperature water bath at 36.5°C for 15 minutes;
[0053] (6) Add 0.1mL thioacetylcholine iodide solution (20mg/mL) and 0.1mL DTNB (10mg/mL) solution to the solution obtained in the previous step and mix well;
[0054] (7) Use a cuvette to compare the color in the 412nm wavelength range, record the absorbance A = 0.9606, and measure the total protein content by the Coomassie brilliant blue method, and then calculate it according to the formula of enzyme activity, specific activity and total enzyme activity, according to enzyme activity Sum specific activity formula calculates that the enzyme activity is 6.1450 U/mL and the specific activity is 0.4836 U/mg.

Example Embodiment

[0055] Example 3:
[0056] (1) Weigh the cleaned and dried watch glass, rinse the fresh chicken heads cut vertically with normal saline, use tweezers to take the chicken brain out of the watch glass, remove the non-brain tissue, and weigh it again to calculate the chicken Brain tissue quality 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 mixture obtained in the previous step with a homogenizer for 2 minutes, let it stand for 30 minutes;
[0059] (4) Centrifuge the homogenate at 8000r/min at 4°C for 20min, and take the supernatant to obtain the crude acetylcholinesterase extract. Place the obtained crude extract in a watch glass and perform 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) Place the supernatant in a watch glass, freeze-dry it in a vacuum with a liquid thickness of 0.6 cm, and collect the acetylcholinesterase product.
[0061] (6) In a 10mL colorimetric tube, add 5mL phosphate buffer (0.1M, pH7.5), then add 0.02mL enzyme solution to mix well, and keep it in a constant temperature water bath at 37°C for 15min;
[0062] (7) Add 0.1mL thioacetylcholine iodide solution (20mg/mL) and 0.1mL DTNB (10mg/mL) solution to the solution obtained in the previous step and mix well;
[0063] (8) Use a cuvette to compare the color in the 412nm wavelength range, record the absorbance A = 0.7119, calculate the enzyme activity to 4.5758 U/mL and the specific activity to 0.3965 U/mg according to the formula of enzyme activity and specific activity.

PUM

PropertyMeasurementUnit
Enzyme activity6.3178U/ml
Specific vitality0.4254U/mg
Specific vitality0.4836U/mg

Description & Claims & Application Information

We can also present the details of the Description, Claims and Application information to help users get a comprehensive understanding of the technical details of the patent, such as background art, summary of invention, brief description of drawings, description of embodiments, and other original content. On the other hand, users can also determine the specific scope of protection of the technology through the list of claims; as well as understand the changes in the life cycle of the technology with the presentation of the patent timeline. Login to view more.

Similar technology patents

Method for preparing high-strength corrugated base paper from straws

ActiveCN107740305AAbundant raw materialsSufficient production
Owner:山东科迈生物制浆有限公司

Classification and recommendation of technical efficacy words

  • Abundant raw materials
  • low cost

Foam concrete

Owner:BEIJING MINJIA NEW BUILDING MATERIALS CO LTD

Preparation of CO2 electro-reduction catalyst, catalyst and application

Owner:QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI

System and method for transmitting wireless digital service signals via power transmission lines

ActiveUS7929940B1reduce bandwidth requirementlow cost
Owner:NEXTEL COMMUNICATIONS

Plastic waveguide-fed horn antenna

InactiveUS20100214185A1low cost
Owner:RGT UNIV OF CALIFORNIA

System and method for determination of position

InactiveUS20090149202A1low costreduce requirement
Owner:STEELE CHRISTIAN

Adaptive antenna optimization network

InactiveUS6961368B2low costminimal space
Owner:ERICSSON INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products