CN100510066C - Superoxide dismutase extraction method - Google Patents
Superoxide dismutase extraction method Download PDFInfo
- Publication number
- CN100510066C CN100510066C CNB2005100227463A CN200510022746A CN100510066C CN 100510066 C CN100510066 C CN 100510066C CN B2005100227463 A CNB2005100227463 A CN B2005100227463A CN 200510022746 A CN200510022746 A CN 200510022746A CN 100510066 C CN100510066 C CN 100510066C
- Authority
- CN
- China
- Prior art keywords
- minutes
- centrifugal
- enzyme
- supernatant liquor
- sod
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The invention relates to a production extracting method of superoxide dismutase, especially to a method for extracting refined copper/zinc superoxide dismutase from a microorganism saccharomyces cerevisiae. It is characterized in that it uses saccharomyces cerevisiae thallium to extract, purify and obtain the refined copper/zinc superoxide dismutase. The craft course is that it obtains the purified refining enzyme by saccharomyces cerevisiae thallium extracting enzyme, acetone secondary precipitation and DEAE-32 fiber column color printing.
Description
Technical field
The present invention relates to the production extracting method of superoxide-dismutase, particularly from a kind of microorganism yeast saccharomyces cerevisiae, extract the extracting method of refining Cu/Zn superoxide dismutase.
Background technology
Superoxide-dismutase (Superoxide dismutise) is called for short SOD, is a kind of metalloenzyme that is prevalent in organism.Mainly be divided into Cu/ZnSOD, Fe-SOD and Mn-SOD according to its bonded metal ion, they form antioxidant enzyme system, catalysis ultra-oxygen anion free radical O jointly with catalase, Selenoperoxidase in body
2 -. disproportionation is H
2O
2And O
2Thereby, stop ultra-oxygen anion free radical to gather in that aerobiont is intravital, eliminate its damage (as Fig. 1) to body.
Fig. 1. biological intravital antioxidase system
CAT: catalase GSH: reduced glutathion
GSSG: Sleep-promoting factor B GSH-PX: Selenoperoxidase
Because special biological function that superoxide-dismutase had, it and HUMAN HEALTH are closely related, can be used for clinically treating ischemic reperfusion syndromes, oxygen intoxication, autoimmune disorder, treat and prevent senile cataract and some inflammation.In addition, SOD also can be used for treating some tetter, as psoriatic, dermatitis, ray photosensitization tetter etc.; In cosmetic applications, superoxide-dismutase has removing beverage, crease-resistant, anti-inflammatory, prevents the function of skin aging; In foodstuffs industry, superoxide-dismutase can be used as efficacy factor or food enrichment is used, and multiple functional food such as existing at present SOD milk, beverage, can, beer appear on the market.
According to patent retrieval, the production method of existing superoxide-dismutase is respectively from animal blood, liver, algae, plant seed such as soybean, extract in the corn etc. and produce, patent as number of patent application 03133482.2, the production method of corn superoxide dismutase from corn is disclosed, the patent disclosure of number of patent application 02150920.4 from marine alga, extract the method for iron super-oxide enzyme, the patent of number of patent application 02147888 relates to a kind of extracting method to superoxide dismutase from pig blood, it is the method for raw material production soybeam superoxides dismutase that number of patent application 921014 discloses with the soybean, above patented method is from plant, extract in the animal and produce superoxide-dismutase, be subjected to raw material aborning, season, production cycle, the influence of factors such as security is bigger, with the microorganism is the method that fermenting raw materials is produced superoxide-dismutase, it is 93103774.3 and the patent of CN01106224.X that number of patent application is only arranged, the former is the wax bacillus, the latter is a milk-acid bacteria, the bacterial classification source is bacterium, the thalline biomass is very low, SOD content is also lower, the former adopts the sedimentary technology of sulphur ammonium could use through desalination, and the SOD of its extraction only is a 1200u/mg albumen than work, and the latter only relates to the fermentation technique of SOD milk-acid bacteria, can only be directly used in the production of sour milk without purifying.
Summary of the invention
The extracting method that the purpose of this invention is to provide a kind of superoxide-dismutase, it is fermenting raw materials production with the microorganism, and culture cycle is short, be easy to large-scale production, and product cost is low, and security and stability are high, and product yield height has higher use value.
The present invention realizes by the following method, and a kind of extracting method of superoxide-dismutase is provided, and it is characterized in that: it extracts purifying with the yeast saccharomyces cerevisiae thalline and obtains Cu/Zn superoxide dismutase.
Described production process with yeast saccharomyces cerevisiae thalline extraction purifying Cu/Zn superoxide dismutase is: the yeast saccharomyces cerevisiae thalline is carried enzyme through broken wall, and acetone precipitation, the DEAE-32 cellulose chromatography obtains the enzyme of purification refine.
It is that centrifugal or Plate Filtration obtained thalline after fermentation by saccharomyces cerevisiae was cultivated that described broken wall is carried enzyme; Discharge enzyme 1:0.8M/V add 45 minutes broken somatic cells walls of 30～35 ℃ of effects of toluene; Under 20～25 ℃ of conditions, with 1:5M/V, add 0.05mol/L pH7.8 phosphoric acid buffer and extracted thick enzyme 4～5 hours, centrifugal 15 minutes of 4000r/min collects supernatant liquor and is crude enzyme liquid; Regulate crude enzyme liquid pH to 4.8～5.0 with 2mol/LHCl again, the centrifugal 15min of 4000r/min collects supernatant liquor, removes the precipitation foreign protein.
Described acetone precipitation; Carry out the acetone primary sedimentation earlier, supernatant liquor is stirred adding 1:0.6V/V cold acetone under salt ice bath condition, left standstill 10 minutes, centrifugal 15 minutes of 4000r/min collects supernatant liquor; Carry out acetone precipitation again: supernatant liquor is regulated about pH to 4.0 with 2mol/LHCl, under salt ice bath condition, continue to stir the cold acetone that adds 1:0.3V/V, centrifugal 15 minutes of 4000r/min, collecting precipitation dissolves precipitation with an amount of 0.05mol/L pH7.8 phosphoric acid buffer.
Described DEAE Mierocrystalline cellulose-32 column chromatography: the DE-32 cellulose column of 1.5cm * 20cm on the solution that step 4 is obtained, with 0.0025mol/L-0.05mol/L, the phosphate buffered saline buffer linear gradient elution of pH7.8, flow rate control are 1ml/3min, and every pipe is collected 3ml; Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid of high vigor.
Described phosphoric acid buffer is the buffered soln that contains 0.1mmol/L EDTA.
Described salt ice bath condition is to add 100 gram trash ices with per 33 gram salt.
Described cold acetone is acetone to be chilled in advance-15 ℃.
Characteristics of the present invention are: because the used bacterial classification of the present invention is a yeast saccharomyces cerevisiae; its biomass be 30～40 the gram about/the L fermented liquid; SOD content is 2000～2600u/g wet thallus; its thalline biomass and SOD content all are higher than other microorganism; culture cycle is short; cost is low, is easy to large-scale production, and Product Safety and stability height.Purification techniques of the present invention is taked acetone fractional precipitation, optionally remove foreigh protein removing, do not need desalinating process, improved purification efficiency, simplified purifying process, and select reasonable parameter to improve yield, the purifying enzyme that purifying obtains reaches 3500u/mg albumen than living, and can be applicable to medical treatment, food, a plurality of fields such as makeup have higher use value.It is the patent of bacterium with respect to the bacterial classification source, patent as number of patent application CN01106224.X is a milk-acid bacteria, number of patent application is 93103774.3 for the wax bacillus, it adopts the sedimentary technology of sulphur ammonium could use through desalination, and the SOD of its extraction only is a 1200u/mg albumen than work.
Description of drawings
The present invention will be further described below in conjunction with the embodiment accompanying drawing.
Accompanying drawing 1 is a yeast saccharomyces cerevisiae superoxide-dismutase extraction and purification process route;
Accompanying drawing 2 is yeast saccharomyces cerevisiae substep purification result;
Accompanying drawing 3 is the active coloring collection of illustrative plates behind the SOD polyacrylamide gel electrophoresis;
Accompanying drawing 4 is SOD enzyme type qualification results.
Embodiment
Embodiment is as shown in Figure 1:
1,1L yeast saccharomyces cerevisiae (former numbering: 2.346, DSMZ of institute of microbiology of the Chinese Academy of Sciences) fermented liquid obtains 40 gram yeast bacterium mud, with distilled water wash twice, and centrifugal collection yeast saccharomyces cerevisiae thalline; Add 32ml toluene with the ratio of 1:0.8 (M/V), 35 ℃ of magnetic agitation broken walls 45 minutes; Add 0.05mol/LpH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA) 200ml with the ratio of 1:5M/V, 25 ℃ of lixiviates 5 hours; Centrifugal 15 minutes of 4000r/min collects the about 180ml of supernatant liquor, is crude enzyme liquid, and total enzyme work is 0.98 * 10
5U;
2, regulate crude enzyme liquid to pH4.8 with 2mol/L HCL, centrifugal 15 minutes of 4000r/min collects the about 170ml of supernatant liquor, and total enzyme work is 0.94 * 10
5U removes the precipitation foreign protein;
3, acetone primary sedimentation: slowly add in advance with the ratio of 1:0.6 (V/V) along walls of beaker and to be chilled to-15 ℃ acetone 102ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (adding 500 gram trash ices) with 165 gram salt.Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min collects the about 260ml of supernatant liquor, and total enzyme work is 0.76 * 10
5U;
4, acetone precipitation: supernatant liquor is regulated about pH to 4.0 with 2mol/LHCl, slowly added in advance with the ratio of 1:0.3 (V/V) along walls of beaker and be chilled to-15 ℃ acetone 78ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (the same).Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min, collecting precipitation is dissolved in 12ml0.05mol/LpH7.8 phosphoric acid buffer (containing 0.1mmol/LEDTA), and total enzyme work is 0.67 * 10
5U;
5, with 1.5 * 20cmDEAE-32 cellulose chromatography on the above-mentioned enzyme liquid, with the phosphate buffered saline buffer linear gradient elution of 0.0025mol/L-0.05mol/LpH7.8, flow rate control 1ml/3min, every pipe is collected 3ml.Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid 18ml of high vigor, total enzyme work is 0.56 * 10
5U.Identify the enzyme purity of superoxide-dismutase at last with polyacrylamide gel electrophoresis; Be accredited as the Cu/Zn superoxide-dismutase through the enzyme type.Its result as shown in Figure 3, the active coloring collection of illustrative plates behind the SOD polyacrylamide gel electrophoresis.Accompanying drawing 4 is SOD enzyme type qualification results, and as shown in Figure 4, the enzyme type of superoxide-dismutase is identified: the purifying enzyme sample is at H
2O
2Act in following 10 minutes inactivation rapidly, KCN handles 10 minutes inactivations about 60%, and chloroform-alcohol does not have influence substantially to this enzymic activity, therefore is accredited as the Cu/Zn superoxide-dismutase.
Embodiment 2
1,1L yeast saccharomyces cerevisiae (former numbering 2.400, DSMZ of institute of microbiology of the Chinese Academy of Sciences) fermented liquid obtains 42 gram yeast bacterium mud, with distilled water wash twice, and centrifugal collection yeast saccharomyces cerevisiae thalline; Add 33.6ml toluene with the ratio of 1:0.8M/V, 32 ℃ of magnetic agitation broken walls 45 minutes; Add 0.05mol/LpH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA) 210ml with the ratio of 1:5M/V, 25 ℃ of lixiviates 4 hours; Centrifugal 15 minutes of 4000r/min collects the about 200ml of supernatant liquor, is crude enzyme liquid, and total enzyme work is 0.88 * 10
5U;
2, regulate crude enzyme liquid to pH5.0 with 2mol/LHCL, centrifugal 15 minutes of 4000r/min collects the about 185ml of supernatant liquor, and total enzyme work is 0.84 * 10
5U removes the precipitation foreign protein;
3, acetone primary sedimentation: slowly add in advance with the ratio of 1:0.6V/V along walls of beaker and to be chilled to-15 ℃ acetone 111ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (adding 500 gram trash ices) with 165 gram salt.Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min collects the about 280ml of supernatant liquor, and total enzyme work is 0.68 * 10
5U;
4, acetone precipitation: supernatant liquor is regulated about pH to 4.0 with 2mol/LHCl, slowly added in advance with the ratio of 1:0.3V/V along walls of beaker and be chilled to-15 ℃ acetone 84ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (the same).Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min, collecting precipitation is dissolved in 10ml0.05mol/L pH7.8 phosphoric acid buffer (containing 0.1mmol/LEDTA), and total enzyme work is 0.60 * 10
5U;
5, with 1.5 * 20cm DEAE-32 cellulose chromatography on the above-mentioned enzyme liquid, with 0.0025mol/L-0.05mol/LpH7.8 phosphate buffered saline buffer linear gradient elution, flow rate control 1ml/3min, every pipe is collected 3ml.Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid 18ml of high vigor, total enzyme work is 0.51 * 10
5U.Identify the enzyme purity of superoxide-dismutase at last with polyacrylamide gel electrophoresis; Be accredited as the Cu/Zn superoxide-dismutase through the enzyme type.
Embodiment 3
1, yeast saccharomyces cerevisiae (former numbering 2.536, DSMZ of institute of microbiology of the Chinese Academy of Sciences) fermented liquid obtains 36 gram yeast bacterium mud, with distilled water wash twice, and centrifugal collection yeast saccharomyces cerevisiae thalline; 1:0.8M/V ratio add 28.8ml toluene, 30 ℃ of magnetic agitation broken walls 45 minutes; Add 0.05mol/L pH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA) 180ml with the ratio of 1:5M/V, 20 ℃ of lixiviates 5 hours; Centrifugal 15 minutes of 4000r/min collects the about 170ml of supernatant liquor, is crude enzyme liquid, and total enzyme work is 0.92 * 10
5U;
2, regulate crude enzyme liquid to pH4.8 with 2mol/LHCL, centrifugal 15 minutes of 4000r/min collects the about 160ml of supernatant liquor, and total enzyme work is 0.88 * 10
5U removes the precipitation foreign protein;
3, acetone primary sedimentation: slowly add in advance with the ratio of 1:0.6V/V along walls of beaker and to be chilled to-15 ℃ acetone 96ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (adding 500 gram trash ices) with 165 gram salt.Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min collects the about 240ml of supernatant liquor, and total enzyme work is 0.70 * 10
5U;
4, acetone precipitation: supernatant liquor is regulated about pH to 4.0 with 2mol/LHCl, slowly added in advance with the ratio of 1:0.3V/V along walls of beaker and be chilled to-15 ℃ acetone 72ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (the same).Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min, collecting precipitation is dissolved in 10ml0.05mol/L pH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA), and total enzyme work is 0.63 * 10
5U;
5, with 1.5 * 20cm DEAE-32 cellulose chromatography on the above-mentioned enzyme liquid, with the phosphate buffered saline buffer linear gradient elution of 0.0025mol/L-0.05mol/L, flow rate control 1ml/3min, every pipe is collected 3ml.Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid 15ml of high vigor, total enzyme work is 0.53 * 10
5U.Identify the enzyme purity of superoxide-dismutase at last with polyacrylamide gel electrophoresis; Be accredited as the Cu/Zn superoxide-dismutase through the enzyme type.
Embodiment 4
1,1L yeast saccharomyces cerevisiae (former numbering: 2.434, DSMZ of Institute of Micro-biology of the Chinese Academy of Sciences) fermented liquid obtains 46 gram yeast bacterium mud, with distilled water wash twice, and centrifugal collection yeast saccharomyces cerevisiae thalline; 1:0.8M/V ratio add 36.8ml toluene, 35 ℃ of magnetic agitation broken walls 45 minutes; Add 0.05mol/LpH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA) 230ml with the ratio of 1:5M/V, 23 ℃ of lixiviates 4.5 hours; Centrifugal 15 minutes of 4000r/min collects the about 210ml of supernatant liquor, is crude enzyme liquid, and total enzyme work is 1.08 * 10
5U;
2, regulate crude enzyme liquid to pH4.8 with 2mol/LHCL, centrifugal 15 minutes of 4000r/min collects the about 200ml of supernatant liquor, and total enzyme work is 1.04 * 10
5U;
3, acetone primary sedimentation: slowly add in advance with the ratio of 1:0.6V/V along walls of beaker and to be chilled to-15 ℃ acetone 120ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (adding 500 gram trash ices) with 165 gram salt.Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min collects the about 280ml of supernatant liquor, and total enzyme work is 0.85 * 10
5U;
4, acetone precipitation: supernatant liquor is regulated about pH to 4.0 with 2mol/LHCl, slowly added in advance with the ratio of 1:0.3V/V along walls of beaker and be chilled to-15 ℃ acetone 84ml, magnetic agitation, walls of beaker overcoat interlayer, interior dress salt ice bath (the same).Left standstill 10 minutes, fully sedimentation, centrifugal 15 minutes of 4000r/min, collecting precipitation is dissolved in 12ml0.05mol/L pH7.8 phosphoric acid buffer (containing 0.1mmol/L EDTA), and total enzyme work is 0.74 * 10
5U;
5, with 1.5 * 20cm DEAE-32 cellulose chromatography on the above-mentioned enzyme liquid, with the phosphate buffered saline buffer linear gradient elution of 0.0025mol/L-0.05mol/L, flow rate control 1ml/3min, every pipe is collected 3ml.Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid 18ml of high vigor, total enzyme work is 0.63 * 10
5U.Identify the enzyme purity of superoxide-dismutase with polyacrylamide gel electrophoresis; Be accredited as the Cu/Zn superoxide-dismutase through the enzyme type.
Above embodiment from yeast saccharomyces cerevisiae substep purification result as shown in Figure 2, extract yeast SOD through above-mentioned used yeast saccharomyces cerevisiae as producing bacterial classification, the SOD yield of purification is 58.3%, be 3500u/mg albumen than living, can be applicable to makeup, health care food production is used, and has certain use value.
Claims (1)
1, a kind of method with yeast saccharomyces cerevisiae thalline extraction purifying Cu/Zn superoxide dismutase, it may further comprise the steps:
1) with after the fermentation by saccharomyces cerevisiae cultivation, centrifugal or Plate Filtration is collected thalline, adds toluene with 1:0.8M/V, and 45 minutes broken somatic cells walls of effect discharge enzyme under 30 ℃～35 ℃ conditions;
2) add the phosphate buffered saline buffer that 0.05mol/LpH7.8 contains 0.1mmol/LEDTA with 1:5M/V, extracted thick enzyme 4～5 hours under 20 ℃～25 ℃ conditions, centrifugal 15 minutes of 4000r/min collects supernatant liquor and is crude enzyme liquid;
3) with step 2) crude enzyme liquid that obtains regulates pH to 4.8～5.0 with 2mol/L HCl, and centrifugal 15 minutes of 4000r/min removes the precipitation foreign protein, collects supernatant liquor;
4) supernatant liquor that step 3) is obtained adds under the salt ice bath condition that 100 gram trash ices form at per 33 gram salt, stir add 1:0.6V/V be chilled to-15 ℃ cold acetone in advance, left standstill 10 minutes, centrifugal 15 minutes of 4000r/min collects supernatant liquor;
5) supernatant liquor that step 4) obtained is regulated pH to 4.0 with 2mol/L HCl, adds under the salt ice bath condition that 100 gram trash ices form at per 33 gram salt, continue to stir add 1:0.3V/V be chilled to-15 ℃ cold acetone, centrifugal 15 minutes of 4000r/min, collecting precipitation in advance;
6) precipitation that step 5) is obtained contains the phosphate buffered saline buffer dissolving of 0.1mmol/L EDTA with the 0.05mol/L pH7.8 of 10mL-12mL, DEAE-32 cellulose column with 1.5cm * 20cm on the solution that obtains after the dissolving, with 0.0025mol/L-0.05mol/L, the phosphate buffered saline buffer linear gradient elution of pH7.8, flow rate control is 1mL/3min, and every pipe is collected 3mL; Through determining the protein quantity and SOD enzyme assay, will contain the active high elutriant of SOD and merge, can obtain the SOD enzyme liquid of high vigor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100227463A CN100510066C (en) | 2005-12-28 | 2005-12-28 | Superoxide dismutase extraction method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100227463A CN100510066C (en) | 2005-12-28 | 2005-12-28 | Superoxide dismutase extraction method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1800377A CN1800377A (en) | 2006-07-12 |
| CN100510066C true CN100510066C (en) | 2009-07-08 |
Family
ID=36810590
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100227463A Expired - Fee Related CN100510066C (en) | 2005-12-28 | 2005-12-28 | Superoxide dismutase extraction method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100510066C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102504000B (en) * | 2011-10-31 | 2014-05-21 | 中兴能源（天津）有限公司 | Method for comprehensively extracting S-adenosine-L-methionine and superoxide dismutase from saccharomyces cerevisiae |
| EP2601965A1 (en) * | 2011-12-06 | 2013-06-12 | Apeiron Biologics AG | Compositions for preventing or treating adverse reactions of EGFR inhibition |
| CN104450635A (en) * | 2014-12-12 | 2015-03-25 | 邢台怡通生物科技有限公司 | Method for extracting and purifying superoxide dismutase (SOD) from saccharomyces cerevisiae |
| CN104593337A (en) * | 2014-12-31 | 2015-05-06 | 唯美度科技（北京）有限公司 | Orgotein extracted from beer yeast |
| CN106987565A (en) * | 2017-05-22 | 2017-07-28 | 大连大学 | A kind of marine low temperature superoxide dismutase extraction and separation process |
-
2005
- 2005-12-28 CN CNB2005100227463A patent/CN100510066C/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 有机溶剂二次沉淀法提取纯化酵母菌超氧化物歧化酶的研究. 杨明琰等.食品科学，第26卷第2期. 2005 |
| 有机溶剂二次沉淀法提取纯化酵母菌超氧化物歧化酶的研究. 杨明琰等.食品科学，第26卷第2期. 2005 * |
| 酵母菌Y－216 SOD的提纯及其性质研究. 王岁楼等.河南大学学报(自然科学版)，第28卷第1期. 1998 |
| 酵母菌Y－216 SOD的提纯及其性质研究. 王岁楼等.河南大学学报(自然科学版)，第28卷第1期. 1998 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1800377A (en) | 2006-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10006001B2 (en) | Methods and compositions to aggregate algae | |
| CN100510066C (en) | Superoxide dismutase extraction method | |
| CN104845896B (en) | Produce the bacterial strain and method of Weilan gum | |
| CN105441525A (en) | Method for increasing yield of haematococcaceae astaxanthin with saccharose as carbon source through co-culture | |
| CN110129251B (en) | Cultivation method of spirulina platensis in salt stress environment | |
| CN106399108A (en) | Simple high-efficiency haematococcus pluvialis nutritive cell culturing and harvesting method | |
| CN113773963A (en) | High-density high-yield porphyridium culture method | |
| US8574887B2 (en) | Process to grow and concentrate algae | |
| CN108485669A (en) | Seaweed activity extract and application thereof | |
| CN1030530C (en) | Preparation of parakin sugar by solidified alpha-glucosyl group transferase | |
| US10745351B2 (en) | Method of producing phycocyanin powder | |
| CN1054398C (en) | Bacillus capable of producing super oxide dismutase and its production method | |
| CN1300311C (en) | Preparation method of chitin incision enzyme | |
| CN1699561A (en) | Process for preparing artemia hatching enzyme by using artemia hatching liquid | |
| CN105969743A (en) | Process for preparing microbial enzyme preparation for degrading formaldehyde | |
| CN102604917A (en) | Preparation method of high-purity nattokinase | |
| CN1062306C (en) | Selection fermentation and extraction technology of brewing yeast metallothionein producing strain | |
| CN101492674A (en) | Method for producing human amyloid A<beta>1-42 with methylotrophy yeast | |
| CN1259427C (en) | Method for preparing grape manna oligosaccharide | |
| JPS6279777A (en) | Production of superoxide dismutase | |
| CN113201067A (en) | Rapid purification cytochrome b6Method of f | |
| CN107090020B (en) | Edible fungus agglutinin and application thereof in inhibiting growth of phaeocystis | |
| RU2237719C2 (en) | Method for preparing lipopolysaccharides | |
| CN102533866A (en) | Antioxidant manufacturing method | |
| CN1259569A (en) | Biotechnology cultivation and processing of lucid ganoderma extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090708 Termination date: 20100128 |