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Method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis

A technology for metallothionein and shrimp scallop, which is applied to the preparation methods, chemical instruments and methods of metallothionein and peptides, can solve problems such as the inability to extract metallothionein at the same time, and achieves high purity, simple extraction process, and improved performance. The effect of economic value

Active Publication Date: 2019-01-15
MARINE FISHERIES RES INST OF ZHEJIANG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purity of metallothionein extracted by this method needs to be improved, and only one kind of metallothionein can be extracted by this method, and two or more pure metallothionein cannot be extracted at the same time

Method used

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  • Method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis
  • Method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis
  • Method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis

Examples

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Embodiment 1

[0034] A method for simultaneously extracting two subtypes of metallothionein from Ezo scallop, comprising the following steps:

[0035] (1) Induction: live scallops were placed in treatment tanks and exposed to Zn + The concentration was 100mg / L zinc chloride solution for 7 days.

[0036] (2) Extraction: take the viscera of the induced scallop, homogenate, add Tris-HCl buffer solution with a concentration of 0.01mol / L, a pH of 8.25, and a temperature of 4°C and a reducing agent TCEP with a concentration of 1.0wt%. , the mass volume ratio of scallop viscera to Tris-HCl buffer solution is 1g / 3mL, overnight at 4°C, centrifuge at 10,000r / min and 4°C for 20min, collect the supernatant a and heat to 70°C for preservation 5min, after rapid cooling, centrifuge, collect the supernatant b, add 3 times the volume of supernatant b ethanol at -20°C, and precipitate overnight; separate the precipitate at 4°C, discard the supernatant, and pour into the precipitate Tris-HCl buffer solution...

Embodiment 2

[0040] A method for simultaneously extracting two subtypes of metallothionein from Ezo scallop, comprising the following steps:

[0041] (1) Induction: live scallops were placed in treatment tanks and exposed to Zn + Concentration of 90mg / L zinc chloride solution induced 8 days.

[0042] (2) Extraction: Take the viscera of the induced scallop, homogenize it, add Tris-HCl buffer solution with a concentration of 0.008mol / L, pH 8, and a temperature of 3°C and a reducing agent TCEP with a concentration of 0.8wt% The mass volume ratio of scallop viscera to Tris-HCl buffer was 1g / 4mL, overnight at 3°C, centrifuged at 12000r / min and 3°C for 17min, collected supernatant a and heated to 75°C for storage 3min, after rapid cooling, centrifuge, collect the supernatant b, add 4 times the volume of supernatant b ethanol with a temperature of -19°C, and precipitate overnight; separate the precipitate at 3°C, discard the supernatant, and pour into the precipitate Tris-HCl buffer was added t...

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Abstract

The invention discloses a method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis, belonging to the technical field of metallothionein extraction. The two subtypes of metallothionein are extracted from the viscera of Patinopecten yessoensis by using the method comprising steps consisting of zinc chloride induction, multiple centrifugation extraction, primary purification via a molecular sieve prepacked column with polymethacrylate as a filling material, and secondary purification via anion exchange chromatography. A mixture of the two subtypes and individual products of the two subtypes obtained by using the method for simultaneously extracting two subtypes of metallothionein from Patinopecten yessoensis in the invention have high purity. Moreover,the extraction process of the method is simple; and the viscera of Patinopecten yessoensis are used as a raw material for the extraction of the metallothionein, and the processing leftovers of enterprises are fully utilized, and the economic value of Patinopecten yessoensis is improved.

Description

technical field [0001] The invention relates to the technical field of metallothionein extraction, in particular to a method for simultaneously extracting two subtypes of metallothionein from Ezo scallop. Background technique [0002] Metallothionein has low molecular weight and high sulfhydryl content, which can bind a large amount of heavy metal ions. It is the most effective free radical scavenger known so far, its ability to scavenge free radicals (·OH) is about thousands of times that of SOD, and the ability to scavenge oxygen free radicals (·O) is about 100 times that of glutathione Peptide (GSH) 25 times, and has a strong antioxidant activity. At present, metallothionein is mostly extracted from rabbit liver, pig liver and plants, and there are few studies on the extraction of metallothionein from aquatic organisms. Since the methods and parameters for extracting metallothionein from different aquatic organisms are not the same, the reference for extracting metallot...

Claims

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Application Information

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IPC IPC(8): C07K14/825C07K1/36C07K1/34C07K1/18C07K1/04
CPCC07K14/825
Inventor 孟春英张小军李停停梅光明
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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