Process for extracting superoxide dismutase (SOD) from houttuynia, hovenia dulcis thumb, rosa roxburghii, myrobalan, bottle gourd and chayote

An extraction process, the technology of Hovenia dulcis, which is applied in the field of extracting high-purity superoxide dismutase, can solve the problems of protein denaturation and inactivation

Inactive Publication Date: 2010-08-25
张卫国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional separation and purification techniques have certain limitations in industrial application, such as step-by-step salting-out and organic solvent precipitation, which can easily cause protein denaturation and inactivation, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The extraction of embodiment 1 Houttuynia cordata high-purity SOD

[0020] A) Take the cleaned Houttuynia cordata and pass through a circulating ultrasonic machine to fully crush it, homogenize it through a homogenizer for about 1 minute, centrifuge at 8000-10000 rpm for 6-30 minutes at high speed, and filter to obtain a crude extract;

[0021] B) the crude extract obtained in step A) is sequentially passed through PEG, GST, and Ni affinity chromatography columns to purify the protein of interest;

[0022] C) adjusting the pH value of the crude extract obtained in step B) to 7-8 through 100-500 mM Tris and 100-500 mM PBS buffer;

[0023] D) the eluent obtained in step C) is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 30 to 50 kDa and a molecular weight cut off of 70 to 150 kDa to prepare high-purity superoxide dismutase, and measure the purity and enzyme activity. The purity of the determination result is as high as 98.2%, and the ...

Embodiment 2

[0027] Example 2 Extraction of Hovenia dulcis Fructus High Purity SOD

[0028] A) Take the clean Hovenia dulcis seeds and fully crush them through a circulating ultrasonic machine, homogenize them with a homogenizer for about 1 minute, centrifuge at 8000-10000 rpm for 10-30 minutes at a high speed, and filter to obtain a crude extract;

[0029] B) the crude extract obtained in step A) is sequentially passed through PEG, GST, and Ni affinity chromatography columns to purify the protein of interest;

[0030] C) adjusting the pH value of the crude extract obtained in step B) to 7.2-7.8 through 100-500 mM Tris and 100-500 mM PBS buffer;

[0031] D) the eluent obtained in step C) is ultrafiltered successively with an ultrafiltration membrane with a molecular weight cut-off of 40 to 50 kDa and a molecular weight cut off of 70 to 150 kDa to prepare high-purity superoxide dismutase, and measure the purity and enzyme activity, The purity of the determination result is as high as 97.8%...

Embodiment 3

[0036] The extraction of embodiment 3 roxburghii high-purity SOD

[0037] A) Take the clean Rosa roxburghii and fully break it through a circulating ultrasonic machine, homogenize it through a homogenizer for about 1 minute, centrifuge at 8000-10000 rpm for 15-30 minutes at a high speed, and filter to obtain a crude extract;

[0038] B) the crude extract obtained in step A) is sequentially passed through PEG, GST, and Ni affinity chromatography columns to purify the protein of interest;

[0039] C) adjusting the pH value of the crude extract obtained in step B) to 7.5-8 through 100-500 mM Tris and 100-500 mM PBS buffer;

[0040] D) the eluent obtained in step C) is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 30 to 50 kDa and a molecular weight cut off of 70 to 150 kDa to prepare high-purity superoxide dismutase, and measure the purity and enzyme activity. The purity of the determination result is as high as 98.5%, and the enzyme activity ...

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Abstract

The invention discloses a process for extracting superoxide dismutase (SOD) from houttuynia, hovenia dulcis thumb, rosa roxburghii, myrobalan, bottle gourd and chayote, which comprises the following steps of: A) preprocessing raw materials, homogenizing the materials by using a homogenizer and centrifuging at the high speed of 8,000 to 10,000 rpm and filtering to produce crude extract; B) allowing the obtained crude extract to pass through polyethylene glycol (PEG), glutathione S-transferase (GST) and nickel (Ni) affinity chromatographic column purified target protein sequentially; C) adjusting the pH value of the crude extract obtained by the step B) to be 7 to 8; D) performing ultrafiltration on the eluent obtained by the step C) by using an ultrafiltration membrane to prepare high-purity SOD; and E) further concentrating the ultrafiltrate to produce SOD concentrated solution and performing freeze drying on the SOD concentrated solution to produce freeze-dried powder for later use. Due to the extracting process, the purity of the separated and purified high-purity SOD can reach 98 percent; the enzyme activity can reach 5.6*10<6>U/g; and the process has high economic value.

Description

technical field [0001] The invention belongs to the technical field of biological enzyme separation and purification engineering, and in particular relates to the extraction of high-purity superoxide dismutase (SOD) from plants such as Houttuynia cordata, Hovenia dulcis, Rosa roxburghii, citronella, gourd, and chayote technology. Background technique [0002] Superoxide dismutase (SOD), as an important scavenger of oxygen free radicals in organisms, can balance oxygen free radicals in the body and avoid adverse reactions caused by excessive concentrations of superoxide anion free radicals in organisms. In radiation protection, diabetes, lung disease, hepatitis, cardiovascular disease, tumor, lupus erythematosus, cataract, rheumatoid, arthritis, degenerative inflammation, exudative inflammation, proliferative inflammation, atopic inflammation and autoimmune therapy, etc. Unique efficacy. At the same time, superoxide dismutase can be used as an active ingredient in beauty co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/08
Inventor 张卫国
Owner 张卫国
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