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Metallothionein (MT) extraction and separation process for marine biological source efficient heavy metal removal agent

A technology of metallothionein and marine organisms, applied in the field of marine biological source metallothionein, can solve the problems of waste of resources, pollution of the environment, etc., and achieve the effect of reducing the impact of safety, reducing industrial costs, and reducing the removal process

Inactive Publication Date: 2014-10-15
国家海洋局第三海洋研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Along with the structural adjustment of my country's aquaculture industry, the aquatic product processing industry has also gradually developed and expanded. However, processed fish products will also produce a large amount of leftovers, accounting for 30% to 50% of processed fish. waste

Method used

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  • Metallothionein (MT) extraction and separation process for marine biological source efficient heavy metal removal agent
  • Metallothionein (MT) extraction and separation process for marine biological source efficient heavy metal removal agent
  • Metallothionein (MT) extraction and separation process for marine biological source efficient heavy metal removal agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Quickly homogenate the fish viscera, a by-product of marine aquatic product processing, weigh 50Kg of the homogenized raw material, put in 350L 0.25 mol / L Tris-HCL buffer solution, and oxidize with hydrochloric acid and hydrogen Sodium controls the pH of the extract to 8.5, and stirs and extracts at 45°C for 2 hours. The extract is heated at 60°C for 60 minutes to remove impurities, and after cooling, the sediment is removed by a continuous flow centrifuge. The centrifugate is first filtered through a plate-and-frame microfiltration membrane Clarify and remove macromolecular compounds at the same time, and then separate, purify and concentrate through ultrafiltration, nanofiltration and other multi-membrane integrated purification systems to obtain crude MT. The crude MT was separated and purified by Sephadex G-50 and DEAE-FF chromatography, and then dried to obtain high-purity marine biological source MT products.

Embodiment 2

[0036]Quickly homogenate the fish viscera, a by-product of marine aquatic product processing, weigh 20Kg of the homogenized raw material, and put in 200L 0.1 mol / L phosphate buffer solution according to the mass volume ratio of the raw material and buffer solution at 1:10, and control it with hydrochloric acid and sodium hydroxide The pH of the extract was 8.0, stirred and extracted at 25°C for 4 hours, the extract was heated at 80°C for 5 minutes to remove impurities, and after cooling, the sediment was removed by a continuous flow centrifuge. The centrifugate was first filtered and clarified through a 0.1um ceramic mold, and Remove macromolecular compounds, and then separate, purify and concentrate with molecular weight cut-offs of 500KDa, 10KDa and 500Da to obtain crude products. The crude MT was separated and purified by Sephadex G-50 and DEAE-FF chromatography, the MT fraction was collected, and then dried to obtain high-purity marine biological source MT products.

Embodiment 3

[0038] Quickly homogenate the fish viscera, a by-product of marine aquatic product processing, weigh 50Kg of the homogenized raw material, put in 250L 0.3mol / L sodium chloride solution according to the mass volume ratio of the raw material and the buffer solution as 1:5, and use hydrochloric acid and sodium hydroxide Control the pH of the extract to 8.5, stir and extract at 55°C for 2 hours, heat the extract at 60°C for 30 minutes to remove impurity proteins, and remove the sediment by a continuous flow centrifuge after cooling, the centrifugate is first filtered and clarified through a plate-and-frame microfiltration membrane , and remove macromolecular compounds at the same time, and then separate, purify and concentrate by molecular weight cut-off of 50KDa and 1KDa respectively to obtain crude MT. The crude SOD was separated and purified by sephadex G-100 and ion exchange resin chromatography, the MT fraction was collected, and then dried to obtain high-purity marine biologi...

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Abstract

The invention provides a metallothionein (MT) extraction and separation process for a marine biological source efficient heavy metal removal agent. The process comprises the following steps: fish viscera subjected to homogenate are added into a buffer solution at the ratio, the pH of an extracting solution is adjusted to 7.0-9.0, the extraction temperature is controlled, and extraction is performed for 1-4 h while stirring; the extracting solution is heated for removing miscellaneous protein, the sediment is removed by centrifugation after cooling, the supernate is subjected to multi-stage ultrafiltration, microfiltration, treated by a nanofiltration multi-component membrane integrated purification system for removing macromolecules, and then subjected to separation and purification, and concentration, so that the MT crude product is obtained; the MT crude product is subjected to separation and purification through the combination of sephadex and ion-exchange column chromatography, and then drying is performed, so that high-purity marine biological source metallothionein product is obtained. The process provided by the invention has the advantages as follows: that the safety is influenced caused by reagent addition in the prior art is reduced, and the subsequent reagent removal process is eliminated; since cheap and rich fish viscera are used as resources for industrial production of MT, the extraction and separation steps are simplified, and a safer efficient marine biological source heavy metal removal agent product is obtained.

Description

technical field [0001] The invention relates to the technical field of high-value development of aquatic products, in particular to the marine biological source metallothionein extracted and separated from fish internal organs, which is a by-product of marine aquatic fish processing, and has an efficient removal effect on heavy metals. Background technique [0002] With the process of industrialization in our country, heavy metals have entered the environment in various ways. When the environment is polluted by heavy metals, they can be enriched in organisms and enter the human body through the food chain, causing acute and chronic poisoning. Heavy metals mainly hinder the absorption of essential trace elements in the human body, interfere with the function of sulfhydryl groups and affect the activity of enzymes and proteins, and have acute toxicity, cumulative toxicity and long-term toxic effects. Long-term consumption of foods with high heavy metal content can easily cause ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/825C07K1/36C07K1/34C07K1/18C07K1/16
CPCC07K14/825
Inventor 孙继鹏王俊坤易瑞灶李诗逸杨婷
Owner 国家海洋局第三海洋研究院
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