CN1087947A - Produce the method for superoxide-dismutase from corn kernel - Google Patents
Produce the method for superoxide-dismutase from corn kernel Download PDFInfo
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- CN1087947A CN1087947A CN 92112213 CN92112213A CN1087947A CN 1087947 A CN1087947 A CN 1087947A CN 92112213 CN92112213 CN 92112213 CN 92112213 A CN92112213 A CN 92112213A CN 1087947 A CN1087947 A CN 1087947A
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Abstract
The invention belongs to the preparation method of enzyme in the biochemical field, relate to the method for from corn, producing superoxide-dismutase.It is characterized in that processing method is: soaking corn; Pulverize; The supernatant liquor ultrafiltration and concentration; Concentrated solution concentrates repeatedly; The centrifugal disgorging of concentrated solution gets supernatant liquor; Supernatant liquor ultrafiltration desalination gets superoxide-dismutase.Technology of the present invention is simple, and is with short production cycle, is suitable for suitability for industrialized production, can provide cheap, safe and reliable superoxide-dismutase product to industries such as medicine, food, daily cosmetics.
Description
The invention belongs to the preparation method of enzyme in the biochemical field, relate to a kind of method invention of from corn, producing superoxide-dismutase.
Nearly more than ten years people have successively developed the work of extracting superoxide-dismutase from the blood of animal and tissue, microorganism, genseng, soybean, and And is applied to medicine, food, cosmetic industry.The superoxide-dismutase of using mainly extracts from animal blood and tissue at present, its cost height, and raw material is not easy to obtain, poor stability, thus limited application.The present invention is to be that raw material extracts with the corn, and the method for similarly producing superoxide-dismutase with the present invention from corn kernel is not seen in the home and abroad by retrieval.
The method purpose that the present invention produces superoxide-dismutase from corn kernel provides a kind of processing method of producing superoxide-dismutase from corn, this superoxide-dismutase is the additive of industries such as medicine, healthcare products, food, daily cosmetics, has medical treatment, anticancer, anti-ageing, the effect of preventing and curing diseases.
The present invention produces the method content Description of superoxide-dismutase from corn kernel:
The present invention a kind ofly produces the method for superoxide-dismutase from corn kernel, it is characterized in that processing method is:
1, soaking corn: after corn kernel cleaned, add soak solution, under 10～50 ℃ of temperature, soaked 20～70 hours, induce the generation superoxide-dismutase;
2, pulverize: the corn kernel after the imbibition adds ammonium sulfate-ammoniacal liquor damping fluid and Sulfothiorine, the dithiothreitol (DTT) of PH=8～8.50.05～0.1 mol of 1.5～3 times and spares prize, stirs then, extracts 0.5～2 hour, and is centrifugal, filters;
3, supernatant liquor ultrafiltration and concentration: high speed centrifugation is removed solid substance, filters then and obtains extracting solution;
4, after concentrated solution adds ammonium sulfate precipitation, centrifugal, the ultrafiltration foreigh protein removing, reconcentration after the desalination, extracting solution is concentrated into 1/4 of original volume by ultra-filtration equipment;
5, concentrated solution is through DE-32, and the P-30 column chromatography for separation adds solid ammonium sulfate to 30-60 degree of saturation and saltouts, and makes most of impurity albumen precipitation, and centrifugal then disgorging gets supernatant liquor;
6, supernatant liquor is further removed foreign protein by ultrafiltration, and ultrafiltration desalination ， And concentrates postlyophilization repeatedly then, gets the elaboration superoxide-dismutase.
The prescription of soak solution is: contain in every premium on currency solution: 412.5 milligrams in ammonium nitrate, 475 milligrams in saltpetre, 42.5 milligrams of potassiumphosphates, 28 milligrams of 7 aqueous ferrous sulfates, 22.3 milligrams of 4 water manganous sulfates, 8.6 milligrams of 7 water zinc sulphates, 0.025 milligram of 5 brochantite, naphthylacetic acid 10
-3PPM, 0.4 milligram of hydrochloric acid thiamine, 0.5 milligram of hydrochloric acid Pyridoxine, 0.5 milligram in nicotinic acid, Plant hormones regulators,gibberellins 10
-1PPM is formulated.
The superoxide-dismutase that the present invention produces the method extraction of superoxide-dismutase from corn kernel can be widely used in industries such as medicine, healthcare products, food and daily cosmetics as additive, have effect anticancer, anti-ageing, beauty treatment, preventable disease is cured the disease, is promoted longevity.From corn, extract superoxide-dismutase than extraction has safe and reliable, virus-free cross infection from animal blood and tissue, raw material sources are extensive, extract characteristics such as having economical rationality than genseng, soybean, residual waste can also fully utilize as producing feed.
Embodiment:
With 1000 gram corn kernels, add 1500 milliliters of soak solutions, after soaking 40 hours under 25 ℃ of temperature, remove soak solution, with the corn kernel of flushing with clean water imbibition three times, add 2000 milliliters of PH=8.2,0.05 the ammonium sulfate of mol-ammoniacal liquor buffered soln, add Sulfothiorine 5 grams, dithiothreitol (DTT) 1 gram simultaneously, at room temperature stir after the pulverizing, extract the centrifugal hypsokinesis of 40 minutes even prize liquid and go out supernatant liquor, filter then, filtrate is that 5000 hollow-fibre ultrafiltration device is concentrated into 500 milliliters by molecular weight cut-off.Under slowly stirring, add solid ammonium sulfate 228 grams several times, make non-superoxide-dismutase albumen precipitation, the elimination precipitation, the filtrate of containing superoxide-dismutase by the molecular retention amount be 100,000 ultra-fine filter again foreigh protein removing be that 5000 filter membrane is concentrated into 100 milliliters with the molecular retention amount then, use 1000 milliliter of 2.5 volumetric molar concentration then respectively, five desalinations of ultrafiltration repeatedly of the potassium phosphate buffer of PH=9.8 get 50 milliliters of concentrated solutions at last.Concentrated solution impouring Whatman DE-32 post (2 * 30 centimetres) is with 5～300 volumetric molar concentrations, and the potassium phosphate buffer of PH=7.8 carries out wash-out, collects active part.Collect Biogel on the liquid, P-30 post (1.5 * 100 centimetres) is with the PH=7.8 of 0.1 mol sodium-chlor, 2.5 the potassium phosphate buffer wash-out of volumetric molar concentration, the collecting high-activity effluent liquid, ultra-fine filter ultrafiltration desalination through molecular retention amount 5000, concentrate postlyophilization, get 20.6 milligrams of superoxide-dismutases, total activity 3080U/ milligram albumen.
The contrast experiment 1
1000 gram corn kernels were soaked 40 hours at 25 ℃ with 1500 ml waters, and the damping fluid that the corn adding of flushing back imbibition is 2000 milliliters, 5 gram Sulfothiorine, 1 gram dithiothreitol (DTT) are pulverized the back stirring and were extracted 40 minutes, and centrifuged supernatant is filtered.
The total activity of embodiment is 150000U;
Contrast experiment 1 total activity is 80000U.
The contrast experiment 2
Do not add under the situation of sodium sulfate, press the described experimental procedure of embodiment and extract superoxide-dismutase, with the embodiment contrast, the embodiment total activity is 112500U;
Contrast experiment 2 total activity is 60000U.
Claims (2)
1, a kind ofly produce the method for superoxide-dismutase, it is characterized in that processing method is from corn kernel:
(1), soaking corn: after corn kernel cleaned, add soak solution, under 10～50 ℃ of temperature, soaked 20～70 hours, induce the generation superoxide-dismutase;
(2), pulverize: the corn kernel after the imbibition adds ammonium sulfate-ammoniacal liquor damping fluid and Sulfothiorine, the dithiothreitol (DTT) of PH=8～8.5,0.05～0.1 mol of 1.5～3 times and spares prize, stirs then, extracts 0.5～2 hour, and is centrifugal, filters;
(3), supernatant liquor ultrafiltration and concentration: high speed centrifugation is removed solid substance, filters then and obtains extracting solution;
(4), after concentrated solution adds ammonium sulfate precipitation, centrifugal, the ultrafiltration foreigh protein removing, reconcentration after the desalination, extracting solution is concentrated into 1/4 of original volume by ultra-filtration equipment;
(5), concentrated solution is through DE-32, the P-30 column chromatography for separation adds solid ammonium sulfate to 30-60 degree of saturation and saltouts, and makes most of impurity albumen precipitation, centrifugal then disgorging gets supernatant liquor;
(6), supernatant liquor further removes foreign protein by ultrafiltration, ultrafiltration desalination repeatedly then, and concentrated postlyophilization, the elaboration superoxide-dismutase.
2, according to claim 1ly produce the method for superoxide-dismutase, it is characterized in that the prescription of said soak solution is: contain 412.5 milligrams in ammonium nitrate, 475 milligrams in saltpetre, 42.5 milligrams of potassiumphosphates, 28 milligrams of 7 aqueous ferrous sulfates, 22.3 milligrams of 4 water manganous sulfates, 8.6 milligrams of 7 water zinc sulphates, 0.025 milligram of 5 brochantite, naphthylacetic acid 10 in every premium on currency solution from corn kernel
-3PPM, 0.4 milligram of hydrochloric acid thiamine, 0.5 milligram of hydrochloric acid Pyridoxine, 0.5 milligram in nicotinic acid, Plant hormones regulators,gibberellins 10
-1PPM is formulated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92112213 CN1087947A (en) | 1992-12-10 | 1992-12-10 | Produce the method for superoxide-dismutase from corn kernel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92112213 CN1087947A (en) | 1992-12-10 | 1992-12-10 | Produce the method for superoxide-dismutase from corn kernel |
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| Publication Number | Publication Date |
|---|---|
| CN1087947A true CN1087947A (en) | 1994-06-15 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 92112213 Pending CN1087947A (en) | 1992-12-10 | 1992-12-10 | Produce the method for superoxide-dismutase from corn kernel |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1060213C (en) * | 1999-01-07 | 2001-01-03 | 王元志 | Method for large scale production of corn superoxide dismutase (SOD) complex enzyme |
| CN1303208C (en) * | 2003-07-23 | 2007-03-07 | 韩伯钧 | Extraction method for SOD complex enzyme coarse powder from corn |
| CN101892204A (en) * | 2010-06-18 | 2010-11-24 | 李作红 | Method for preparing corn superoxide dismutase from corn |
| CN103173423A (en) * | 2013-03-29 | 2013-06-26 | 湖南大学 | Method for extracting superoxide dismutase with high yield from corn |
| CN101553570B (en) * | 2006-10-09 | 2014-07-30 | 法国植物基因组研究计划--华莱 | Improvement of the kernel productivity of plants through the modulation of glutamine synthetase activity |
-
1992
- 1992-12-10 CN CN 92112213 patent/CN1087947A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1060213C (en) * | 1999-01-07 | 2001-01-03 | 王元志 | Method for large scale production of corn superoxide dismutase (SOD) complex enzyme |
| CN1303208C (en) * | 2003-07-23 | 2007-03-07 | 韩伯钧 | Extraction method for SOD complex enzyme coarse powder from corn |
| CN101553570B (en) * | 2006-10-09 | 2014-07-30 | 法国植物基因组研究计划--华莱 | Improvement of the kernel productivity of plants through the modulation of glutamine synthetase activity |
| CN101892204A (en) * | 2010-06-18 | 2010-11-24 | 李作红 | Method for preparing corn superoxide dismutase from corn |
| CN103173423A (en) * | 2013-03-29 | 2013-06-26 | 湖南大学 | Method for extracting superoxide dismutase with high yield from corn |
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