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Industrial extraction and purification method for rabbit liver zinc metallothionein

A zinc metallothionein and rabbit liver technology, which is applied in the extraction and purification of metallothionein, can solve problems such as unsatisfactory results, achieve the effects of poor mechanical strength of colloidal particles, enhance tolerance and adaptability, and reduce losses

Inactive Publication Date: 2007-12-12
HUNAN LUGU BIOTECH
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AI Technical Summary

Problems solved by technology

Five international conferences on MT have been held in the world, and the fifth international conference on MT was held in Beijing last year. , its use is also increasingly extensive, so the demand for MT is also increasing. Although there are many reports about the method of extracting MT on a large scale, the results are not ideal. Therefore, it is necessary to provide a large-scale, High-extraction, low-cost production process for extracting metallothionein

Method used

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Experimental program
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Effect test

Embodiment

[0019] 1. Induction

[0020] Choose healthy adult cyanine rabbits about 3kg, and use the prepared ZnSO 4 ·7H 2 O solution was induced by subcutaneous injection in the abdomen, injected every other day, a total of 7 inductions, each dose was 10mg / kg, 20mg / kg, 50mg / kg, 100mg / kg, 150mg / kg, 200mg / kg, 250mg / kg; On the fifteenth day, the rabbits were slaughtered and the livers were taken.

[0021] 2. Rough extraction

[0022] Wash the removed liver with normal saline, weigh 6000g, mince the rabbit liver and put it into the extraction tank-1, add 12L of 0.02M (pH 8.6) Tris-HCl buffer, stir to homogenate, and extract For 30 minutes, add 6L of ethanol: chloroform = 1:0.08 denaturing solution, centrifuge at 4°C, 4000 rpm for 30 minutes, discard the precipitate, take the supernatant, obtain supernatant 1, add supernatant 1 to the extraction In the tank-2, the temperature of the extraction tank-2 was raised to 75°C rapidly, kept for 5 minutes, and then quickly lowered to room temperat...

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Abstract

The invention relates to an industrial method for extracting and purifying zinc metallothionin from rabbit liver. It comprises following steps: inducing, coarse extracting, chromatography purifying; inducing adult rabbit for 5- 7 times through belly hypodermic injection, the zinc injection dosage is increased by 10- 250 mg for every kg rabbit gradually; stirring rabbit liver with homogenate liquor, adding denatured liqour and stirring, freezing and centrifuging, collecting supernatant -1, heating fastly, colling fastly to normal temperature, freezing and centrifuging again, getting supernatant -2, condensing through ultra- filtering membrane, getting supernatant -3 for chromatography in gel medium, desalting collecting liquid and freeze drying. The invention is characterized by higher productivity of Zn- MT, low extracting cost, high purity, simple preparation process and convenient extracting method.

Description

technical field [0001] The present invention relates to the extraction and purification method of metallothionein, in particular to the extraction and purification method of rabbit liver 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. MT molecule is elliptical, with a molecular weight of 6500 Daltons and a diameter of 30-50A. It is divided into two structural domains. Each molecule contains 7-12 metal atoms and has special light absorption. The MT conformation is stronger and has stronger heat resistance. Typical mammalian MTs have a molecular weight ...

Claims

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Application Information

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IPC IPC(8): C07K14/825C07K1/14C07K1/34
Inventor 江冠群蒋腊才王少波曹醴承
Owner HUNAN LUGU BIOTECH
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