CN1896256A - Method for converting resveratrolside into resveratrol by microbion - Google Patents
Method for converting resveratrolside into resveratrol by microbion Download PDFInfo
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- CN1896256A CN1896256A CN 200610200633 CN200610200633A CN1896256A CN 1896256 A CN1896256 A CN 1896256A CN 200610200633 CN200610200633 CN 200610200633 CN 200610200633 A CN200610200633 A CN 200610200633A CN 1896256 A CN1896256 A CN 1896256A
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Abstract
A method for converting resveratrolside into resveratrol by microbe is carried out by crushing raw material, inducing carbon source and nitrogen source to obtain seed culture medium and fermentative culture medium, inoculating Aspergillus oryzae, inducing gas, agitating, fermenting and extracting fermentative liquid by microwave. It is simple and cheap, has shorter production period and more output.
Description
Technical field
The invention belongs to biotransformation of Natural Products and extractive technique field, specially refer to the method that the polydatin in the plant is converted into trans-resveratrol with microorganism.
Background technology
Trans-resveratrol is a kind of resveratrol compounds, is present in the various plants such as grape, black false hellebore, Tuber Fleeceflower Root, giant knotweed, have anticancer, antiviral, anti-oxidant, stop effect such as platelet aggregation, be mainly used in prevention and treatment cardiovascular disorder.At present, trans-resveratrol has been widely used in health care of food product and makeup, and its demand is rising year by year.
The production method of trans-resveratrol has chemosynthesis and extracts from natural phant.Chemical synthesis cost height, contaminate environment is so the production of trans-resveratrol mainly is to extract from plant at present.Yet the Resveratrol content in the plant is low especially, and wherein content is the highest in giant knotweed, but also only reaches 0.43% (area, Hanzhong), and this just makes that its separation and purification cost is higher.And the content of resveratrol analogs white hellebore glycosides in plant is far above trans-resveratrol.In giant knotweed, the content of polydatin can reach 2.5% (area, Hanzhong); In Tuber Fleeceflower Root, its high-content can reach 4.8% (Sichuan).If polydatin is converted into trans-resveratrol, the productive rate of trans-resveratrol can improve greatly.
Traditionally, the acquisition of aglycon realizes by the corresponding glucosides of hydrolysis.But hydrolysis reaction need be finished by acid or base catalysis under High Temperature High Pressure, to the equipment requirements height, and environment is polluted.By contrast, biotransformation method has mild condition, and process is simple, pollutes advantages such as little, has become the important means of drug modification.
Polydatin is converted into trans-resveratrol mainly to be realized by enzymatic conversion method and microbial transformation.For example, (CN1251363 FR2795964), perhaps adds cellulase be hydrolyzed (CN1566349A) in the giant knotweed extracting solution to add prozyme constant temperature enzyme digestion reaction 48-72 hour in Powdered giant knotweed raw material.Though enzymatic conversion method has obtained certain effect, enzyme costs an arm and a leg, and has limited promoting the use of of these methods.Microbe transformation method is that the giant knotweed after soaking into is placed temperature is spontaneous fermentation 24-96 hour (CN1385535A) or add special microorganism at giant knotweed and transform (CN1513822A) in 20-50 ℃ the container.Microbe transformation method can improve the productive rate of trans-resveratrol effectively, but the flora the unknown in the spontaneous fermentation process, condition is difficult to control, and airborne mould might produce toxic substance; Microorganism used therefor is unexposed in the special microorganism conversion method (CN1513822A), can't verify and repeat, and its practicality is subjected to very big restriction.
Summary of the invention
The purpose of this invention is to provide a kind of food-grade microorganisms aspergillus oryzae (Aspergillus oryzae) as bacterial classification, raw material giant knotweed, Tuber Fleeceflower Root, Stem of Smalleaf Jointfir, black false hellebore, Testa arachidis hypogaeae, Pericarpium Vitis viniferae or Semen Vitis viniferae etc. are transformed, carry out microwave-assisted again and extract, can improve the yield of trans-resveratrol greatly.
For achieving the above object, processing step of the present invention comprises:
The preparation of substratum: seed culture medium is the bran mass of the raw material powder of interpolation 0 ~ 6%.Must possess required carbon source of microorganism growth and nitrogenous source in the fermention medium.Carbon source is the wheat bran of 10 ~ 30g/L, and nitrogenous source is the ammonium sulfate of 5 ~ 9g/L, the ammonium nitrate of 3 ~ 7g/L or the urea of 3 ~ 5g/L.Substratum must be sterilized down at 121 ℃ and can be used in 15 ~ 20 minutes.
Seed culture: carry out in shaking bottle, shaking speed is 100 ~ 200rpm, and temperature is 20 ~ 35 ℃, and incubation time is 18 ~ 24h.
Fermentation culture: carry out in the 5L fermentor tank, inoculum size is 5% ~ 15%, and rotating speed of agitator is 150 ~ 350rpm, and temperature is 20 ~ 40 ℃.The bubbling air amount is 0.8 ~ 1.5vvm in the fermenting process, and fermentation time is 12 ~ 48 hours.After fermentation was carried out 16 ~ 44 hours, changing culture temperature was 45 ~ 55 ℃, continued to cultivate 1 ~ 6 hour.
Product extracts: add one times 50 ~ 95% ethanol in the fermented liquid and carry out microwave-assisted and extract.
The invention has the advantages that directly raw material powder fermented that improved the yield of its trans-resveratrol greatly, technology is simple, good reproducibility.Microorganism used therefor is food grade, and is safe and reliable.
Embodiment
Below in conjunction with technical scheme, be example with the giant knotweed, be described in detail embodiments of the present invention.
Embodiment 1:
The used bacterial classification that ferments is aspergillus oryzae (Aspergillus oryzae), available from Chinese industrial microbial strains preservation center, is numbered CICC 2436.
Substratum: A. seed culture medium: the 10g wheat bran adds 100ml boiling tap water 20min, filters, and filtrate adds water to 100ml, adds 6% Giant Knotweed Rhizome again.B. fermention medium: Giant Knotweed Rhizome 200g (60 order), wheat bran 50g, ammonium nitrate 12.5g, tap water 2500ml, 121 ℃ of sterilization 20min.
Fermenting process: seed culture is carried out in the triangular flask of 250ml, and liquid amount is 30mL, and shaking speed is 150rpm, is culture temperature 30?, incubation time is 24 hours; Fermentation culture is carried out in the 5L fermentor tank, and actual liquid amount is 2.5L, and inoculum size is 5%, and rotating speed is controlled to be 250rpm, and the bubbling air amount is 1.5vvm, is temperature controlled at 37?
Fermentation ends adds 1 times to 95% ethanol of fermented liquid, and microwave-assisted extracts.
The result is as follows for the experiment gained:
Fermentation was carried out 24 hours, and the productive rate of trans-resveratrol is 1.34% (g/g medicinal material), was 3.12 times of trans-resveratrol productive rate (0.43%) in the medicinal material of not fermenting.
Embodiment 2:
The used bacterial classification that ferments is aspergillus oryzae (Aspergillus oryzae), available from Chinese industrial microbial strains preservation center, is numbered CICC 2436.
Substratum: A. seed culture medium: the 10g wheat bran adds 100ml boiling tap water 20min, filters, and filtrate adds water to 100ml.B. fermention medium is with embodiment 1.
Fermenting process: seed culture is carried out shaking in the bottle of 250ml, and liquid amount is 30mL, and shaking speed is 150rpm, is culture temperature 25?, incubation time is 24 hours; Fermentation culture is shaken in the bottle at 250ml and is carried out, and actual liquid amount is 50ml, and inoculum size is 5%, and rotating speed is controlled to be 150rpm, is temperature controlled at 25?Was changing culture temperature 50 after fermentation was carried out 44 hours?, continue to cultivate 2 hours.
After cultivating end, add 95% ethanol that is doubled in fermented liquid, microwave-assisted extracts.
The result is as follows for the experiment gained:
The productive rate of trans-resveratrol is 1.35% (g/g medicinal material), is 3.86 times of trans-resveratrol productive rate (0.35%) in the medicinal material of not fermenting.
Claims (8)
1. one kind is converted into the method for trans-resveratrol with microorganism with polydatin, it is characterized in that processing step comprises:
(1), adds carbon source and nitrogenous source, preparation seed culture medium and fermention medium with raw material pulverizing;
(2) inoculation aspergillus oryzae;
(3) stir fermentation;
(4) microwave-assisted extracts fermented liquid.
2. according to claim 1ly a kind ofly polydatin is converted into the method for trans-resveratrol, it is characterized in that described raw material is giant knotweed, Tuber Fleeceflower Root, Stem of Smalleaf Jointfir, black false hellebore, Testa arachidis hypogaeae, Pericarpium Vitis viniferae or Semen Vitis viniferae with microorganism.
3. according to claim 1ly a kind ofly with microorganism polydatin is converted into the method for trans-resveratrol, it is characterized in that inoculating aspergillus oryzae is Aspergillus oryzae (CICC 2436), is food-grade microorganisms.
4. according to claim 1ly a kind ofly polydatin is converted into the method for trans-resveratrol, it is characterized in that seed culture medium is the bran mass that adds 0 ~ 6% raw material powder with microorganism.
5. a kind of method that polydatin is converted into trans-resveratrol with microorganism according to claim 1, it is characterized in that adding in the fermention medium wheat bran that carbon source is 10 ~ 30g/L, nitrogenous source is the ammonium sulfate of 5 ~ 9g/L, the ammonium nitrate of 3 ~ 7g/L or the urea of 3 ~ 5g/L.
6. a kind of method that polydatin is converted into trans-resveratrol with microorganism according to claim 1, it is characterized in that inoculum size is 5 ~ 15%, the fermentor tank mixing speed is 150 ~ 350rpm, culture temperature is 20 ~ 40 ℃, air feeding amount is 0.8 ~ 1.5vvm, and fermentation time is 12 ~ 48 hours.
7. according to claim 6ly a kind ofly with microorganism polydatin is converted into the method for trans-resveratrol, after its feature was that also fermentation is carried out 16 ~ 44 hours, changing culture temperature was 45 ~ 55 ℃, continues to cultivate 1 ~ 6 hour.
8. according to claim 1ly a kind ofly polydatin is converted into the method for trans-resveratrol, it is characterized in that it is 50 ~ 95% ethanol that microwave-assisted extracts the fermented liquid solvent for use with microorganism.
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| CN2006102006332A CN1896256B (en) | 2006-06-29 | 2006-06-29 | Method for converting resveratrolside into resveratrol by microbion |
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| CN1896256A true CN1896256A (en) | 2007-01-17 |
| CN1896256B CN1896256B (en) | 2010-07-28 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104894172A (en) * | 2014-03-05 | 2015-09-09 | 财团法人食品工业发展研究所 | Method for producing resveratrol by microbial transformation and Brussels Dekkera mutant strain |
| CN108033874A (en) * | 2017-12-28 | 2018-05-15 | 长沙善道新材料科技有限公司 | Biological extraction method of resveratrol |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090603C (en) * | 1999-09-24 | 2002-09-11 | 四川省凉山州生物研究所 | Process for extracting resveratrol from Chinese medicine giant knotweed root |
| CN1116264C (en) * | 2000-07-20 | 2003-07-30 | 北京孚曼生物技术有限公司 | Method for separating reseveratrol from resveratrol glucoside and application thereof |
| CN1341587A (en) * | 2001-09-17 | 2002-03-27 | 上海汇谷生物技术有限公司 | Method for preparing resveratrol from giant knotweed rhizome |
-
2006
- 2006-06-29 CN CN2006102006332A patent/CN1896256B/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104894172A (en) * | 2014-03-05 | 2015-09-09 | 财团法人食品工业发展研究所 | Method for producing resveratrol by microbial transformation and Brussels Dekkera mutant strain |
| CN108033874A (en) * | 2017-12-28 | 2018-05-15 | 长沙善道新材料科技有限公司 | Biological extraction method of resveratrol |
| CN108033874B (en) * | 2017-12-28 | 2020-11-20 | 亓云吉 | Biological extraction method of resveratrol |
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