CN1626654A - Method for distilling superoxide dismutase from corn - Google Patents
Method for distilling superoxide dismutase from corn Download PDFInfo
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- CN1626654A CN1626654A CN 200310119109 CN200310119109A CN1626654A CN 1626654 A CN1626654 A CN 1626654A CN 200310119109 CN200310119109 CN 200310119109 CN 200310119109 A CN200310119109 A CN 200310119109A CN 1626654 A CN1626654 A CN 1626654A
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Abstract
A process for extracting SOD from corn incldues disinfecting corn grains, germinating, beating, squeezing to obtain juice, depositing, centrifugal separation to remove dregs, ultrafiltration, extracting by active peak, concentrating and freeze drying. Said SOD features high activity, low content of protein, and high resistance to hydrolysis.
Description
Technical field
What the present invention relates to is a kind of preparation method of biological enzyme, in particular the preparation method of superoxide-dismutase (SOD).
Background technology
Superoxide-dismutase is called for short SOD again, is a kind of scavenging agent of important oxyradical, has very strong anti-peroxy radical ability, and it is widely used in healthcare products, medicine and the makeup.Present SOD extracts from animal blood, its extracting method is complex steps not only, and to consume a large amount of industrial chemicals, produce the large quantity of exhaust gas polluted air, simultaneously the chloroform that contains in its waste liquid, acetone, phosphoric acid etc. all are normal organic substances of imitating, and can't reclaim, and its discharging, permeate the ground and will directly cause phreatic acidifying, alkalization to pollute, and carry pathogenic micro-organism in the blood raw material, country also strictly limits in rules and regulations.It is big that blood extracts the SOD molecular weight, is difficult to permeate through cell membranes, also easily by Pepsin or Trypsin hydrolysis, has active more weak defective in the application, can only under 0-4 ℃ of temperature environment, survive usually, otherwise inactivation.Though also disclose with the corn in the prior art is the method that raw material extracts SOD, and its method is different, also has the big disadvantage of industrial chemicals consumption.Chinese patent CN99112607, name are called " corn superoxide dismutase from corn (SOD) prozyme method for large scale production ", its method is to utilize corn, through germinate, add phosphoric acid salt, pull an oar, add cellulase, pull an oar, add calcium chloride, transition, add the stove enzyme, diatomite filtration, centrifugal, ultrafiltration, vitaminize C, vitamin-E, potassium sorbate, freezing, melt centrifugal a series of process and produce superoxide-dismutase, this technical scheme exists complex process, technology disadvantage that the industrial raw material consumption is big equally, still has the technological deficiency that is difficult to scale operation.
Summary of the invention
The for a change existing blood that still relies on merely is the state of the art that raw material extracts rising of super-oxide enzyme extraction approach, and have now with the corn is that rising of raw material extraction super-oxide enzyme exists the technological deficiency that produces a large amount of polluting wastes, it is the technical scheme that raw material extracts the extracting method of superoxide-dismutase with the corn that patent application of the present invention provides a kind of, and the major technique content of the technical program is:
1. corn kernel sterilization, germination
Adjust room temperature, water temperature, pH value, humidity, oxygen feed rate, illumination, substratum etc. to good basis that is fit to that corn kernel sprouts and environment;
2. pull an oar, press juice
The inclusion-free pH value that adds one times of weight ratio by the germination kernel weight is the deionized water of 6-7, making beating, and juice is got in squeezing then, precipitation, its thick slag of elimination;
3. post precipitation is centrifugal removes slag;
The buffer reagent that adds its volume percent 15% in the juice that pressure is got stirs, and thin slag and starch are staticly settled, and gets its supernatant liquor after centrifugal, extracts clear liquid and sends into ultrafiltration;
4. filtrate ultrafiltration
Clear liquid is carried out ultrafiltration, and its clear liquid is sent into extraction column;
5. extract active peak
Clear liquid adds promotor, impels its accelerate precipitation, sends into extraction column, after some grades of diffusions, reclaims throw out;
6., concentrate the distillation freeze-drying:
Throw out is concentrated, and it is freezing in the freeze-drying dish that the extract after concentrating is packed into;
Its 3. the buffer reagent in the step be: the citric acid solution of the ionized water volume 35% that accounts for and the sodium citrate solution that accounts for dilution water volume 35% are mixed and made into; Its 5. the promotor in the step be acetone and 95% ethanol respectively by volume per-cent add with in supernatant liquor at 85%: 15%.
What patent application of the present invention provided is a whole set of extracting method that raw material extracts superoxide-dismutase with the corn, its raw material sources are abundant, extracting method is simple, be more suitable for big industrial production, and its molecular weight of SOD that present method is extracted only be 1/2 of blood SOD molecular weight, overcome blood SOD molecular weight big, be difficult for permeate through cell membranes also easily by the defective of Pepsin, Trypsin hydrolysis.The SOD that this method is extracted is not only a kind of of SOD ferment enzyme, it also exists Agifutol, catalase, peroxidase simultaneously, it is a kind of prozyme of multienzyme kind, can keep the activity of SOD more longways, can resist Pepsin, tryptic hydrolysis.Because it originates from plant sugar line style cell, so it contains V
E, V
B, V
CDeng various trace elements such as multivitamin and copper, iron, zinc, manganese, selenium, and these elements and VITAMIN all are indispensable material and nutrition in the human body.Its final product is creamy white, and has pink colour less.Assert that after testing corn SOD is the most complete at present, the most a kind of ferment enzyme in removing human body aspect the free radical, can not be substituted.This enzyme family belongs to acidic protease family, and is activity stabilized, and thermotolerance is arranged, and can keep its activity under 15-25 ℃ temperature environment, and especially some performance is very stable in PH7.6～9, showing less stable below the PH6 or more than 12.SOD compares with blood:
| Hundred kilograms of rate of recovery (g) | Vigor (U/mgprot) | Protein content (mgpot/ml) | |
| Blood SOD | ?20～25 | ?400～500 | ?0.541 |
| Corn SOD | ?100～110 | ?700～1000 | ?0.047 |
The SOD final product that corn extracts is a zymin.It has strengthened the lupus erythematosus of treatment general; rheumatoid arthritis; dermatitis; scleroderma; autoimmune hemolytic anemia; thrombocytopenia; autoimmune disorder; myocardial ischemia; the ischemia-reperfusion syndromes; cardiovascular disorder; kidney; liver; the protection of organs such as heart and transplanting; the replantation of a severed limb; repair behind the cancer chemotherapy; maintenance prevents the diffusion and the oxidation of cancer cells; prevent the formation of cancer cells; beauty treatment; surgical procedures such as shaping are anti-oxidation; anti-aging; the damage of antiultraviolet irradiation; external application can reach skin-protecting face nursing; removing beverage and cosmetic result.
Embodiment
To describe the technology contents of the technology of the present invention below in detail:
1, the SOD of corn extraction mainly is taken from the line style cell caryoplasm of corn bud sugar, so its matter of utmost importance is the quality and the percentage of germination of corn.In its preparation step process,, at first to clean and sterilization for removing undiscovered crushed particles and the mushroom that its crust of elimination may carry in the corn kernel; The cultivation of germinateing then: pH value, air flow quantity, illumination and the oxygen-supplying amount of adjusting its envrionment temperature, humidity, water by the corn germination condition, because ferment family all belongs to happiness oxygen bacterium, adjust substratum, reach the oxygen supply of budding period necessity, give illumination in 24 hours simultaneously, infrared rays and ultraviolet light light fixture can be increased, not only the budding photosynthesis that sprouts can be helped, simultaneously also can be to the germicidal action of space, chamber.When corn grows duckbill yellow-green colour sprout, just can carry out rinsing and pulverize.
2, making beating, pressure are squeezed the juice
Adding weight ratio by the actual weight of the corn that sprouts is 1: 1 inclusion-free deionized water, and the pH value of its deionized water preferably is adjusted between the 6-6.5, breaks into pulping then; The full slurry of accomplishing fluently is put in the squeezing machine, with dynamicliquid pressure juice is all extruded, it presses juice to send into next treatment step; Its thick slag can be sent into proving room, uses for ferment wine brewing.
3, post precipitation is centrifugal removes slag:
In the juice that pressure is got, slowly add buffer reagent, under condition of stirring, add 15% of juice volume, its buffer reagent is: the sodium citrate solution of the citric acid solution of the ionized water volume 65% that accounts for and the ionized water volume 35% that accounts for is mixed and made into, stir simultaneously, make its actives be suspended state, staticly settle then, its supernatant liquor is after what rev/mins 30 minutes are centrifugal, extract its clear liquid, and uf processing in the pumping ultrafiltration jar.
4, filtrate ultrafiltration
Through the differential screen separatory, its separatory is after ultrafiltration in ultra-filtration equipment, and its ultrafiltration clear liquid send extraction column again;
5, extract active peak liquid
In clear liquid, add promotor, its promotor be acetone and 95% ethanol respectively by volume per-cent add with in supernatant liquor at 85%: 15%, send into extraction column, this extraction column is made as level Four, liquid is removed in diffusion, reclaims throw out, and throw out is sent into concentration tank concentrates.Contained acetone, ethanol is all recyclable in the clear liquid of emitting after the extractions at different levels utilizes again.
6, concentrate the distillation freeze-drying:
Throw out is sent into concentration tank carry out concentration, the semi-fluid after concentrating pack into distillation in the freeze-drying dish, freeze-drying obtain the SOD crude product.
Each step after the 6th step is in order to produce medical SOD elaboration, and the processing step that carry out is:
7, the SOD crude product after will concentrating dilutes with deionized water, with water-bath and dialysis dialyzate is sent into the centrifugal liquid that removes of whizzer, reclaims its throw out, with the freeze-drying of throw out reconcentration, promptly obtains elaboration SOD, and its freeze-drying packing is consistent with the SOD crude product.
Claims (4)
1, a kind of extracting method of corn superoxide dismutase from corn is characterized in that its extracting method is:
1. corn kernel sterilization, germination
Adjust room temperature, water temperature, pH value, humidity, oxygen feed rate, illumination, substratum etc. to good basis that is fit to that corn kernel sprouts and environment;
2. pull an oar, press juice
The inclusion-free pH value that adds one times of weight ratio by the germination kernel weight is the deionized water of 6-7, making beating, and juice is got in squeezing then, precipitation, its thick slag of elimination;
3. post precipitation is centrifugal removes slag:
The buffer reagent that adds its volume percent 15% in the juice that pressure is got stirs, and thin slag and starch are staticly settled, and gets its supernatant liquor after centrifugal, extracts clear liquid and sends into ultrafiltration:
4. filtrate ultrafiltration
Clear liquid is carried out ultrafiltration, and its clear liquid is sent into extraction column;
5. extract active peak liquid
Clear liquid adds promotor, impels its accelerate precipitation, sends into extraction column, after some grades of diffusions, reclaims throw out;
6., concentrate the distillation freeze-drying;
Throw out is concentrated, and the extract after concentrating is packed into, and the freeze-drying dish distils, freeze-drying;
Its 3. the buffer reagent in the step be: the sodium citrate solution of the citric acid solution of the ionized water volume 65% that accounts for and the ionized water volume 35% that accounts for is mixed and made into; Its 5. the promotor in the step be acetone and 95% ethanol respectively by volume per-cent add with in supernatant liquor at 85%: 15%.
2, extracting method according to claim 1,2. the step is 6-6.5 with the pH value of deionized water to it is characterized in that described the.
3, extracting method according to claim 1 is characterized in that 4. it the go on foot its clear liquid earlier through the differential screen separatory, carries out ultrafiltration again.
4, extracting method according to claim 1, it is characterized in that for obtaining elaboration SOD the the SOD crude product after 6. will concentrating after the step dilutes with deionized water, it is centrifugal except that liquid with water-bath and dialysis dialyzate to be sent into whizzer, reclaim its throw out, with the freeze-drying of throw out reconcentration, packing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310119109 CN1261565C (en) | 2003-12-11 | 2003-12-11 | Method for distilling superoxide dismutase from corn |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310119109 CN1261565C (en) | 2003-12-11 | 2003-12-11 | Method for distilling superoxide dismutase from corn |
Publications (2)
| Publication Number | Publication Date |
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| CN1626654A true CN1626654A (en) | 2005-06-15 |
| CN1261565C CN1261565C (en) | 2006-06-28 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200310119109 Expired - Fee Related CN1261565C (en) | 2003-12-11 | 2003-12-11 | Method for distilling superoxide dismutase from corn |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421750A (en) * | 2012-05-23 | 2013-12-04 | 王兆非 | Industrial production method for preparing superoxide dismutase compound enzymes with corn as raw material and sprouting room |
| CN103740667A (en) * | 2014-01-15 | 2014-04-23 | 李美凤 | Extraction method of corn SOD enzyme through ionic liquid |
| CN109845594A (en) * | 2016-05-30 | 2019-06-07 | 路平 | A kind of implantation methods of multienzyme crops containing SOD and its application |
| CN111041007A (en) * | 2019-12-04 | 2020-04-21 | 上海润日生物科技发展有限公司 | Preparation method of superoxide dismutase |
-
2003
- 2003-12-11 CN CN 200310119109 patent/CN1261565C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421750A (en) * | 2012-05-23 | 2013-12-04 | 王兆非 | Industrial production method for preparing superoxide dismutase compound enzymes with corn as raw material and sprouting room |
| CN103421750B (en) * | 2012-05-23 | 2018-12-28 | 王兆非 | Industrialized preparing process and its rudiment room of superoxide dismutase complex enzyme are prepared using corn as raw material |
| CN103740667A (en) * | 2014-01-15 | 2014-04-23 | 李美凤 | Extraction method of corn SOD enzyme through ionic liquid |
| CN109845594A (en) * | 2016-05-30 | 2019-06-07 | 路平 | A kind of implantation methods of multienzyme crops containing SOD and its application |
| CN111041007A (en) * | 2019-12-04 | 2020-04-21 | 上海润日生物科技发展有限公司 | Preparation method of superoxide dismutase |
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| Publication number | Publication date |
|---|---|
| CN1261565C (en) | 2006-06-28 |
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