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Extraction method of sheep embryo active peptide

An extraction method, sheep embryo technology, applied in the field of extraction of embryonic active peptides, can solve the problems of many by-products, damaged amino acids, and restrictions on wide application, and achieve the effects of avoiding pollution, maintaining biological activity, and avoiding environmental pollution

Inactive Publication Date: 2013-08-28
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The preparation of biologically active peptides by the acid-base method is simple, but it has the disadvantages of serious damage to amino acids, difficulty in controlling the degree of hydrolysis, many by-products, affecting the structure and function of the peptide, and there may be acid, alkali or other substances remaining, so it is limited. Wide application of this method
The enzymatic hydrolysis method causes more loss of peptides and amino acids, which reduces the activity of active peptides; and the process is more cumbersome and the cost is higher
Other methods of extracting small molecular peptides mainly cannot recover all the active peptides required, and it is difficult to achieve the purpose of large-scale production

Method used

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  • Extraction method of sheep embryo active peptide
  • Extraction method of sheep embryo active peptide
  • Extraction method of sheep embryo active peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation and detection of sheep embryo active peptide solution

[0021] Take 30g of fresh thymus tissue of 6-week-old sheep embryo and chop it up, add 30ml of normal saline, and homogenize it with a tissue homogenizer; after homogenization, put it in a -20°C refrigerator for quick freezing, take it out after 30 minutes, and put it in a 40°C water bath for rapid melt. Ultrasonic lysis, set the frequency band of ultrasound to 3000Hz, start the ultrasound probe every 30 seconds, and the duration of ultrasound is 15 seconds, repeat this step 3 times, so that all the small molecular active peptides in the cells are released. Centrifuge at a high speed of 15,000g for 30 minutes at a low temperature of 4°C to remove larger impurity fragments that cannot be broken by ultrasonic cracking in the tissue homogenate, and obtain a clear and transparent supernatant. Ultrafiltration of the supernatant, using a Microcon YM-10 (0.22 μm pore size) ultrafiltration device for ...

Embodiment 2

[0025] Example 2 Preparation of sheep embryo active peptide freeze-dried powder

[0026] Take 80 g of new liver, thymus, and spleen tissue of 16-week sheep embryo, chop them up, add 50 ml of normal saline, and homogenize with a tissue homogenizer. After homogenization, put it in a -20°C refrigerator for quick freezing, take it out after 30 minutes, put it in a 40°C water bath for quick thawing, put it into a -20°C refrigerator for quick freezing after melting, take it out after 30 minutes, and put it in a 40°C water bath Melt again quickly. For ultrasonic lysis, set the frequency band to 3000 Hz, start the ultrasonic probe at intervals of 30 seconds, and the duration of ultrasound is 15 seconds, repeat the ultrasonic lysis step 5 times, so that all the small molecular active peptides in the cells are released. Centrifuge at a low temperature of 15,000 g for 30 minutes at a low temperature at 4°C to remove larger impurity fragments that cannot be broken by ultrasonic cracking...

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Abstract

The invention discloses an extraction method of sheep embryo active peptide, which sequentially comprises the following steps of: (1) fetching the fresh tissue of 6-16 week sheep embryo, cutting up and adding normal saline; homogenizing by a tissue homogenizing machine; and performing quick freeze thawing at -20 DEG C and 40 DEG C after the homogenization; (2) performing ultrasonic cracking: starting an ultrasonic probe every 30 seconds, letting the ultrasound last for 15 seconds, and repeating for 3-5 times; (3) performing low-temperature high-speed centrifugation for 30 minutes at 4 DEG C, and taking the supernate; and (4) performing centrifugal ultrafiltration of the supernate by use of a 5k ultrafiltration membrane, and clarifying to obtain the ultrafiltrate which is a sheep embryo active peptide solution. Without any acid and alkali treatment, the extraction method disclosed by the invention effectively avoids environmental pollution caused by a chemical method, and also avoids product pollution. Moreover, the method can collect multiple biologically active peptide components to the greatest degree, wherein the molecular weight of the active peptide is effectively controlled below 5,000D, and the bioactivity of the sheep embryo small-molecular peptide is maintained to the greatest degree.

Description

technical field [0001] The invention relates to a method for extracting biologically active substances, in particular to a method for extracting embryonic active peptides. Background technique [0002] Sheep embryo small molecular bioactive peptide, also known as sheep placenta, is the regulatory element of embryonic cell differentiation and development, can make fertilized eggs accurately differentiate into various tissue cells, and can promote cell differentiation and cell proliferation. Small molecule active peptides are highly conserved, have high homology, no immunogenicity, no allergic reactions, no side effects on the human body, and can be used by humans. Small molecule bioactive peptides can directly enter cells to repair damaged cells. Damage the DNA of cells, protect and repair damaged cells, and restore the structure and function of damaged cells. Therefore, the sheep embryo small molecule bioactive peptide can be used to activate the vitality of human cells, re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14
Inventor 张建湘张李洋唐岸柳何国平
Owner CENT SOUTH UNIV
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