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Method for preparing zinc metallothionein

A zinc metallothionein and production method technology, which is applied in the direction of metallothionein and peptide preparation methods, chemical instruments and methods, etc., can solve the complex process, affect the metallothionein purification precision and nutritional effect and other problems, and achieve the speed of proposal fast effect

Inactive Publication Date: 2006-07-19
黄世宽
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only the process is complicated, but also because the inducer used in the induction process is ZnCL 2 , so there must be a desalination process, otherwise it will seriously affect the purification accuracy and nutritional effect of metallothionein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Take 5 kg of fresh rabbit liver containing zinc metallothionein, and wash it repeatedly with distilled water in the raw material washing process to remove all impurities.

[0013] In the raw material crushing process, the washed rabbit liver is put into a pulverizer for crushing, passed through a 60-200 mesh sieve, and then put into a container.

[0014] In the homogenizing process, the pulverized raw material is stirred with the solvent to achieve thorough mixing. The used solvent is ethanol solution, and the content of ethanol is between 60% and 85%. The weight ratio of the ethanol solution to the raw material is 2-4:1, and the main function of the ethanol is to leach the zinc metallothionein in the raw material and make it enter the solution.

[0015] In the filtration process, the homogenate is filtered by a centrifuge to retain the liquid and remove the residue. Or place the homogenate in a filter cloth for natural filtration, and get the filtrate.

[...

Embodiment 2

[0017] Example 2. Get 5Kg of fresh rabbit liver containing zinc metallothionein. In the raw material cleaning process, first wash and disinfect with normal saline to remove impurities, and then wash repeatedly with distilled water to remove normal saline and all impurities.

[0018] In the raw material crushing process, the washed rabbit liver is put into an ultrasonic pulverizer for crushing, and the crushed raw materials are put into a container.

[0019] In the homogenization process, the pulverized raw materials are stirred with the solvent to achieve full mixing. The used solvent is ethanol solution, and the content of ethanol is 65%. The weight ratio of ethanol solution to raw material is 2:1.

[0020] In the filtration process, the homogenate is filtered by a centrifuge, the liquid is retained, and the residue is removed. Or place the homogenate in a filter cloth for natural filtration, and get the filtrate.

[0021] In the extraction process, the liquid is first vac...

Embodiment 3

[0022] Example 3. The raw material cleaning process, the raw material crushing process, and the filtering process are the same as in Example 2.

[0023] In the homogenization process, the ethanol solution used has an ethanol content of 75%. The weight ratio of ethanol solution to raw material is 2.5:1.

[0024] In the extraction process, the still temperature in a vacuum state is 50°C ± 2°C; the temperature of the freezer used for the freeze separation is carried out in a freezer at -30°C, and kept for 6 hours, the precipitate is retained, and the liquid (mainly is ethanol) recovered for reuse.

[0025] The precipitate, whose main component is zinc metallothionein, was subjected to chromatographic separation. In the chromatographic separation process, the precipitate is mixed with distilled water at a ratio of 1:3-4.5. Then put the solution into the SephardexG-50 column for chromatography, MT-1 stays in the column, MT-2 flows into the container outside the column, then washe...

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PUM

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Abstract

The disclosed preparation method for Zn-metalloprotein comprises: cleaning and crushing material for homogenate; filtering, vacuum distilling in kettle of 35-60Deg for extraction; condensing, froze separating at -90~-10Deg, and vacuum drying. This invention has high extraction and purification rate with short period and simple process.

Description

Technical field: [0001] The invention relates to a production method of zinc metallothionein, belonging to one of the production methods of metallothionein. Background technique: [0002] In 1957, when Margoshes and Vallee were studying the biological effects of metals, a new protein was isolated from animal organs. It contains rich sulfhydryl groups and can chelate a large amount of metal ions. This substance is called metallothionein (Metallothionein in English, Abbreviated as MT). Human body, animals, plants and microorganisms all contain MT, and their physical and chemical properties are basically the same. The molecular weight is 6500-7000 Daltons, the diameter is 30-50A, it is divided into two structural domains, each molecule contains 7-12 metal atoms, and has special light absorption. It contains 61 amino acids, 21 of which are cysteine, so that each MT molecule can bind up to seven metal atoms. MT is chemically stable and can survive in water at 80°C for 30 minut...

Claims

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Application Information

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IPC IPC(8): C07K14/825C07K1/14
Inventor 黄世宽
Owner 黄世宽
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