CN104388478A - Method for improving yield of resveratrol through fermentation of aspergillus niger - Google Patents
Method for improving yield of resveratrol through fermentation of aspergillus niger Download PDFInfo
- Publication number
- CN104388478A CN104388478A CN201410681191.2A CN201410681191A CN104388478A CN 104388478 A CN104388478 A CN 104388478A CN 201410681191 A CN201410681191 A CN 201410681191A CN 104388478 A CN104388478 A CN 104388478A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- resveratrol
- aspergillus niger
- giant knotweed
- trans
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/22—Preparation of oxygen-containing organic compounds containing a hydroxy group aromatic
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for improving yield of resveratrol through fermentation of aspergillus niger. An aspergillus niger strain is cultured in a triangular flask culture medium for 48h, 2% by mass of hypha is evenly added to giant knotweed rhizomes with humidity of 2:1, and the stacking fermentation is carried out at a constant temperature of 40 DEG C for 30h. Through a solid stacking fermentation method, polydatin in giant knotweed rhizomes is converted into resveratrol, and the solid fermentation condition is optimized. In the method, a strong acid-base reaction is not generated, and glycoside does not need to be crudely extracted; meanwhile, expensive equipment does not need to be added, and the method is easy to popularize. The extraction yield of resveratrol is increased to 1.197% and is increased by 30% compared with that of the existing method, so the very obvious technical effects are achieved.
Description
Technical field
The invention belongs to field of microbial fermentation, relate to the application of aspergillus niger, be specifically related to a kind of method utilizing fermentation of Aspergillus niger to improve trans-resveratrol yield.
Background technology
The effective constituent of giant knotweed Polygonumcuspidatum is mainly diphenylethylene and anthraquinone analog compound, and the former is Resveratrol and piceid mainly, and the latter is Schuttgelb and glucosides thereof mainly, wherein has significantly unique effect with diphenylethylene compounds.Trans-resveratrol has Tumor suppression, anti-oxidant, Green Tea Extract, antithrombotic, antianaphylaxis, atherosclerosis and has preventive and therapeutic effect to coronary heart disease, ischemic heart disease, hyperlipidemia etc., and trans-resveratrol has been listed in one of anti-the most promising cardiovascular, anticancer medicine.It is reported, in dry giant knotweed rhizome, the content of trans-resveratrol is 0.1% ~ 0.2%, and Determination of Polydatin is about 2%, and how efficiently polygonin to be converted into trans-resveratrol becomes study hotspot.
At present, have people's acid and alkali hydrolysis or modified-cellulose enzymolysis process to decompose polygonin, but soda acid can cause environmental pollution, reduce output, and biological enzymolysis cost is higher, technique is comparatively complicated, is therefore all unfavorable for applying.Also have and adopt submergence fermentation mode microbes producing cellulase direct converting giant knotweed glycosides to be the report of trans-resveratrol, but equipment and operational requirement high, capital construction has high input.
Chinese patent 201110374767.7 discloses a kind of aspergillus niger, deposit number: CCTCC No:M 2011358, and the Resveratrol in Rhizoma Polygoni Cuspidati content of its bio-conversion processes, reaches 0.92%.But the method also has the space promoted further.
Summary of the invention
Given this, the object of the invention is to provide a kind of method that technique is simple, use fermentation of Aspergillus niger that is that be easy to promote improves trans-resveratrol yield.
Solve above technical problem, technical scheme provided by the invention is, a kind of method using fermentation of Aspergillus niger to improve trans-resveratrol yield is provided, aspergillus niger strain is put in triangular flask substratum and cultivate 48h, in mass percent be 2% ratio mycelia is evenly added in the moistening giant knotweed rhizome for 2:1, heap fermentation 30h under 40 DEG C of constant temperature.
Further, described substratum is based on czapek's solution, removes sucrose wherein, adds the mistake 80 object giant knotweed powder that mass percent is 1%.
Compared with prior art, a technical scheme tool in technique scheme has the following advantages:
1, the present invention adopts solids bulk fermentation method that polygonin in giant knotweed is changed into resveratrol, and is optimized solid fermentation condition.This method had not both had strong acid-base reaction to occur, and again without the need to slightly carrying glycosides, needing to add expensive equipment simultaneously, being easy to promote.
2, trans-resveratrol of the present invention extracts yield and is increased to 1.197%, improves 30%, achieve the technique effect of highly significant than existing method productive rate.
Embodiment
Be described below in conjunction with specific embodiment.
In Aspergillus niger strain heap fermentation converting giant knotweed, the condition of polygonin mainly contains: temperature, inoculum size, inoculation cell age, fermentation humidity.
Inoculum size, cell age, humidity are fixed as respectively 2%, 36h, 2:1, leavening temperature is set as 30,40,50,60 DEG C of 4 gradients, carries out heap fermentation, transforms the situation of trans-resveratrol every 6h sampling analysis polygonin.Earlier fermentation polygonin conversion rate with leavening temperature rise and accelerate, transforming degree also improves thereupon, but temperature higher than 40 DEG C time, Resveratrol content ferment 30h time reach peak value after, after progressively decline.
The bacterial strain body endoenzyme system developmental state that cell age is different is different, and the ability conformed is also different.Inoculum size, temperature, humidity are fixed as respectively 2%, 40 DEG C, 2:1, cell age are set as 24,36,48,60h, 4 gradients, carry out heap fermentation, transform the situation of trans-resveratrol every 6h sampling analysis polygonin.For mould liquid culture, fermentation 24h often bacterial strain logarithmic growth early stage, ferment 60h then thalline enter the secondary metabolism initial stage, because of Medium Proportion and fermentation condition change, slightly different.When being 24h strain fermentation by cell age, maximum Resveratrol content is only 0.75%, and is 60h strain fermentation by cell age, although bacterium endoenzyme system concentration is high, bacterium reproduction speed is slow, causes transforming degree to be the bacterium of 36h and 48h lower than cell age.The breeding of comprehensive bacterium and bacterium enzymatic productivity, select to be that the bacterium of 48h inoculates and carries out heap fermentation into giant knotweed and be advisable by cell age.
Cell age, temperature, humidity are fixed as respectively 48h, 40 DEG C, 2:1, fermentation inoculum size is set as 1%, 2%, 3%, 4%4 gradients, carries out heap fermentation, and in 6h sampling analysis giant knotweed, polygonin transforms the situation of trans-resveratrol.The impact of inoculum size in heap fermentation process is as liquid-immersed fermentation, and namely inoculum size is little, then the lag phase of fermenting extends, and means that production intensity reduces and fermentation microbiological contamination risk improves to fermentation; But inoculum size is excessive, mean that cost also increases as seed culture increase, therefore inoculum size selects 2% to be advisable.
Inoculum size, temperature, cell age are fixed as respectively 2%, 40 DEG C, 48h, the ratio that water and giant knotweed dry mass are added in fermentation is set as 1.8:1.2:1,2.2:1,2.5:1 totally 4 gradients, heap fermentation, in 6h sampling analysis giant knotweed, polygonin transforms the situation of trans-resveratrol.In fermenting process, water activity increases, and namely humidity improves, and contributes to thalli growth and secretes the diffusion of enzyme outward, and therefore humidity increase can accelerate enzymolysis and the raising trans-resveratrol extraction yield of polygonin; But then, humidity improves the energy consumption but increasing trans-resveratrol leaching process, makes separation and purification process cost increase.Thus when heap fermentation, humidity must have one just when.Found through experiments, when humidity is increased to 2:1 by 1.8:1, it is larger that Resveratrol content increase rate is produced in fermentation; And when improving humidity further, Resveratrol content amplification slows down.Therefore, selection humidity is 2:1, and giant knotweed fermentation is ideal.
Put by aspergillus niger strain in triangular flask substratum and cultivate 48h, substratum is based on czapek's solution, removes sucrose wherein, adds the mistake 80 object giant knotweed powder that mass percent is 1%.Grow well until mycelia, mix with suitable quantity of water and giant knotweed rhizome.The mass ratio of water and giant knotweed rhizome is 2:1, and the inoculation amount for mycelia is 2% of giant knotweed dry weight.Heap fermentation 30h under 40 DEG C of constant temperature.
Fermentation sample extraction trans-resveratrol time, the giant knotweed that first air seasoning ferments, after add quantitative ethanol, thermostatically heating extract certain hour, suction filtration.Get a certain amount of extracting solution and use ether, extraction into ethyl acetate respectively, be separated and obtain trans-resveratrol, polygonin.
Adopt ultraviolet spectrophotometry to do quantitative analysis, trans-resveratrol has maximum absorption at 306nm, polydatin at 325nm place.After extracting solution is separated with organic solvent extraction, measures absorbance in the maximum absorption wave strong point of each effective constituent respectively with 756MC type ultraviolet-visible pectrophotometer, calculated the mass concentration of each effective constituent by typical curve, thus calculate extraction yield.By calculate, trans-resveratrol extract yield be increased to 1.197%, this extraction yield compared with in prior art 0.92% extraction yield improve 30%, achieve the technique effect of highly significant.
Claims (2)
1. the method using fermentation of Aspergillus niger to improve trans-resveratrol yield, it is characterized in that, being put by aspergillus niger strain in triangular flask substratum and cultivate 48h, is that mycelia is evenly added to moistening in the giant knotweed rhizome of 2:1 by the ratio of 2% in mass percent, heap fermentation 30h under 40 DEG C of constant temperature.
2. use fermentation of Aspergillus niger according to claim 1 improves the method for trans-resveratrol yield, and it is characterized in that, described substratum is based on czapek's solution, removes sucrose wherein, adds the mistake 80 object giant knotweed powder that mass percent is 1%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410681191.2A CN104388478A (en) | 2014-11-24 | 2014-11-24 | Method for improving yield of resveratrol through fermentation of aspergillus niger |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410681191.2A CN104388478A (en) | 2014-11-24 | 2014-11-24 | Method for improving yield of resveratrol through fermentation of aspergillus niger |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104388478A true CN104388478A (en) | 2015-03-04 |
Family
ID=52606435
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410681191.2A Pending CN104388478A (en) | 2014-11-24 | 2014-11-24 | Method for improving yield of resveratrol through fermentation of aspergillus niger |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104388478A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109503334A (en) * | 2018-12-21 | 2019-03-22 | 广东冠龙生物科技有限公司 | A kind of preparation process preparing 99% resveratrol of purity from polygonum cuspidate |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030086986A1 (en) * | 1998-08-06 | 2003-05-08 | Bruijn Chris De | Ophthalmic, pharmaceutical and other healthcare preparations with naturally occurring plant compounds, extracts and derivatives |
| CN102533565A (en) * | 2011-11-22 | 2012-07-04 | 浙江工业大学 | Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed |
-
2014
- 2014-11-24 CN CN201410681191.2A patent/CN104388478A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030086986A1 (en) * | 1998-08-06 | 2003-05-08 | Bruijn Chris De | Ophthalmic, pharmaceutical and other healthcare preparations with naturally occurring plant compounds, extracts and derivatives |
| CN102533565A (en) * | 2011-11-22 | 2012-07-04 | 浙江工业大学 | Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed |
Non-Patent Citations (1)
| Title |
|---|
| 王卫等: "虎杖固体发酵生产白藜芦醇工艺条件优化研究", 《林产化学与工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109503334A (en) * | 2018-12-21 | 2019-03-22 | 广东冠龙生物科技有限公司 | A kind of preparation process preparing 99% resveratrol of purity from polygonum cuspidate |
| CN109503334B (en) * | 2018-12-21 | 2022-04-12 | 广东冠龙生物科技有限公司 | Preparation process for preparing resveratrol with purity of 99% from polygonum cuspidatum |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Rodríguez et al. | Bioethanol production from grape and sugar beet pomaces by solid-state fermentation | |
| CN103981054B (en) | A kind of biological enzyme brewages the method for oil tea wine | |
| CN104893983B (en) | Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product | |
| CN103352062A (en) | Method for preparing glycyrrhetinic acid monoglucuronide | |
| CN101736033A (en) | Method for producing red yeast rice with functions of regulating lipoid and reducing blood pressure through submerged fermentation | |
| CN103431368A (en) | Preparation method and comprehensive utilization of submerged fermentation product of cordyceps sinensis liquid | |
| CN108753900A (en) | A kind of method that the fermentation of ginseng endophyte prepares rare ginsenoside | |
| CN101928671A (en) | Method for preparing ginsenoside Rg3 from alternaria alternata YS-2 and fermented ginseng stem-leaf total saponin thereof | |
| CN107174598B (en) | Method for comprehensively utilizing overground stem and leaf parts of salvia miltiorrhiza | |
| CN101376669B (en) | Preparation of 6-O-beta-D- glucosyl-3,6,16,25-tetrahydroxy cycloartane | |
| CN102363796A (en) | Method for producing glycyrrhetinic acid through microbial fermentation transformation | |
| CN104388478A (en) | Method for improving yield of resveratrol through fermentation of aspergillus niger | |
| CN106148203A (en) | A kind of Absidia coerulea and application technology thereof | |
| CN103554287A (en) | Extraction method of boletus edulis mycelium polysaccharide | |
| CN102242159B (en) | Method for improving resveratrol content in giant knotweed by two-step microbial conversion | |
| CN105462857A (en) | Marasmiellus androsaceus strain and application thereof | |
| CN106834407B (en) | Method for green production of diosgenin by biological method | |
| CN101812483B (en) | Method for converting polydatin into resveratrol by coupling steam explosion and biological methods | |
| CN109022501A (en) | A method of ferulic acid is obtained using waste | |
| CN104278070A (en) | Method for improving content of ergosterol in liquid fermentation products of phellinus igniarius | |
| CN101307337B (en) | Process for preparing coenzyme Q10 by sphingosine unit cell strain fermentation, extraction and coupling | |
| CN107723316A (en) | A kind of method that resveratrol yield is improved using fermentation of Aspergillus niger | |
| CN106399397A (en) | Method for increasing content of tyrosol in rhodiola rosea by microorganism fermentation | |
| CN103865803B (en) | Beta-glucosidase Producing Strain and the application prepared in genipin and trans-resveratrol in conversion thereof | |
| CN105296545A (en) | Method for converting polydatin through microorganism to prepare resveratrol |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150304 |
|
| WD01 | Invention patent application deemed withdrawn after publication |