CN103695478A - Method for converting polydatin to resveratrol - Google Patents
Method for converting polydatin to resveratrol Download PDFInfo
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- CN103695478A CN103695478A CN201310720687.1A CN201310720687A CN103695478A CN 103695478 A CN103695478 A CN 103695478A CN 201310720687 A CN201310720687 A CN 201310720687A CN 103695478 A CN103695478 A CN 103695478A
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- resveratrol
- endogenetic fungus
- substratum
- polydatin
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Abstract
The invention discloses a method for converting polydatin to resveratrol. The method comprises the following steps: (1) preparing an endophytic fungi L1 spore suspension; (3) preparing a culture medium; and (3) fermenting. The method is simple in fermentation condition, easy to control, strong in enzymatic conversion specificity, high conversation rate, and low in cost and environmental-friendly.
Description
Technical field
The present invention relates to a kind of method that polydatin is converted into trans-resveratrol, especially relate to a kind of microbial method that utilizes and polydatin is converted into the method for trans-resveratrol.
Background technology
Trans-resveratrol is a kind of natural radioactivity polyphenolic substance, has to suppress tumour, protect the multiple pharmacological effect such as cardiovascular, anti-oxidant, Green Tea Extract, is listed in one of cardiovascular disease resistant, anticancer the most promising medicine.
At present, the method for preparing trans-resveratrol mainly comprises: 1. plant extraction method, and low conversion rate, cost is high.2. chemical synthesis, synthesis step is various, and trans-resveratrol is very easily oxidized, and reaction yield is restricted; Many catalyzer and chemical reagent that building-up process is used all have certain hazardness, and environmental pollution is serious.3. culture plant cell method, the reason such as the inherited character of high yielding cell sarain is unstable, is difficult to realize rapidly large-scale industrial production.As can be seen here, aforesaid method cuts both ways.Find a kind of transformation efficiency high, the trans-resveratrol preparation method that cost is low, has become the task of top priority of current trans-resveratrol production industry.
Summary of the invention
The technical problem to be solved in the present invention is, overcomes the deficiencies in the prior art, provides a kind of transformation efficiency high, and the polydatin that cost is low is converted into the method for trans-resveratrol.
The technical scheme that the present invention solves its technical problem employing is: a kind of polydatin is converted into the method for trans-resveratrol, comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is the patent application document that CGMCC NO.4558(is 201110130127.1 referring to application number);
(2) substratum is prepared: fresh giant knotweed is crushed to 200 orders, in mass ratio giant knotweed 1.5wt%, dehydrated alcohol 2 wt%, CMC-Na 2.5 wt%, NH
4nO
30.2 wt%, Na
2hP0
40.06 wt%, CaCl
20.1 wt% preparation fermention medium, tune Initial pH is 4.5-5.5;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; Again by endogenetic fungus L1 spore suspension, access in the substratum of initial pH 4.5-5.5, inoculum size is the 5-10%(preferably 9% that is equivalent to culture volume), in 250ml triangular flask, the liquid amount of substratum is 30mL, under culture condition is temperature 26-30 ℃, rotating speed 200-220r/min, fermentation 36-60h.
HPLC detects polydatin and Resveratrol content in fermented liquid, and transformation efficiency >=60% transforms Resveratrol content >=30 u g/mL in secondary fermentation liquid.
It is trans-resveratrol that the present invention's microbial method transforms polydatin, first separated from fresh giant knotweed, be purified into endogenetic fungus L1, then according to the growth properties feature of L1, prepare specific substratum, under certain culture condition, by L1 converting giant knotweed polydatin, be trans-resveratrol, guarantee the high-content of trans-resveratrol, and then improve the output of trans-resveratrol.Polydatin is exactly the moiety of giant knotweed, and the trans-resveratrol that traditional extraction can only be extracted in giant knotweed can not extract polydatin, and the present invention is exactly to be trans-resveratrol by the polydatin in converting giant knotweed, and then improves output.
Technique fermentation condition of the present invention is simple, easy to control, and enzymatic conversion specificity is strong, and transformation efficiency is high, and cost is low, environmentally friendly, is one of preferred approach of the natural trans-resveratrol of manufacture; And can realize suitability for industrialized production, environmental pollution is little, resource environment is friendly, is to produce the important channel that the high value of natural white lamb's-quarters sound alcohol develops.
Embodiment
Below in conjunction with embodiment, the invention will be further described.
Embodiment 1
The present embodiment comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is the patent application document that CGMCC NO.4558(is 201110130127.1 referring to application number);
(2) substratum preparation: fresh giant knotweed is crushed to 200 orders, gets the fresh Giant Knotweed Rhizome 0.45g(of 200 order giant knotweed 1.5wt%), dehydrated alcohol 0.6g(2%), CMC-Na0.75g(2.5%), NH
4nO
30.06g(0.2%), Na
2hP0
40.018g(0.06%), CaCl
20.03g(0.1%) be mixed with fermention medium; Adjusting initial pH is 5.5;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; By in the endogenetic fungus L1 spore suspension access 250ml triangular flask of 2.7ml, the substratum of the initial pH 5.5 of 30mL is housed in described triangular flask, then is under 28 ℃ of temperature, rotating speed 220r/min at culture condition again, fermentation 60h.
HPLC detects polydatin and Resveratrol content in fermented liquid, and obtaining the transformation efficiency that polydatin is converted into trans-resveratrol is 95.6%, transforms Resveratrol content in secondary fermentation liquid and reaches 52.533 u g/mL, is unfermentable 9.90 times.
Embodiment 2
The present embodiment comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is the patent application document that CGMCC NO.4558(is 201110130127.1 referring to application number);
(2) substratum preparation: get the fresh Giant Knotweed Rhizome 0.45g(of 200 order giant knotweed 1.5%), dehydrated alcohol 0.6g(2%), CMC-Na0.75g(2.5%), NH
4nO
30.06g(0.2%), Na
2hP0
40.018g(0.06%), CaCl
20.03g(0.1%) be mixed with fermention medium; Adjusting initial pH is 4.5;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; Again by the endogenetic fungus L1 spore suspension access 250ml triangular flask of 1.4ml, in described 250ml triangular flask, be equipped with in the 250ml triangular flask of initial pH 4.5 substratum of 20mL, at culture condition, be under 26 ℃ of temperature, rotating speed 200r/min again, fermentation 36h.
HPLC detects polydatin and Resveratrol content in fermented liquid, and obtaining polydatin, to be converted into trans-resveratrol transformation efficiency be 60.2%, transforms Resveratrol content in secondary fermentation liquid and reach 33.080 u g/mL, is unfermentable 6.23 times.
Embodiment 3
The present embodiment comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is the patent application document that CGMCC NO.4558(is 201110130127.1 referring to application number);
(2) substratum preparation: get the fresh Giant Knotweed Rhizome 0.45g(of 200 order giant knotweed 1.5%), dehydrated alcohol 0.6g(2%), CMC-Na0.75g(2.5%), NH
4nO
30.06g(0.2%), Na
2hP0
40.018g(0.06%), CaCl
20.03g(0.1%) be mixed with fermention medium; Adjusting initial pH is 5.0;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; Again the endogenetic fungus L1 spore suspension access of 3.6ml is equipped with in the 250ml triangular flask of initial pH 5.0 substratum of 40mL, then is under 28 ℃ of temperature, rotating speed 200r/min at culture condition, fermentation 48h.
HPLC detects polydatin and Resveratrol content in fermented liquid, and obtaining polydatin, to be converted into trans-resveratrol transformation efficiency be 77.8%, transforms Resveratrol content in secondary fermentation liquid and reach 42.752 u g/mL, is unfermentable 8.06 times.
Embodiment 4
The present embodiment comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is the patent application document that CGMCC NO.4558(is 201110130127.1 referring to application number);
(2) substratum preparation: get the fresh Giant Knotweed Rhizome 0.45g(of 200 order giant knotweed 1.5%), dehydrated alcohol 0.6g(2%), CMC-Na0.75g(2.5%), NH
4nO
30.06g(0.2%), Na
2hP0
40.018g(0.06%), CaCl
20.03g(0.1%) be mixed with fermention medium; Adjusting initial pH is 4.5;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; Again the endogenetic fungus L1 spore suspension access of 2.7ml is equipped with in the 250ml triangular flask of initial pH 4.5 substratum of 30mL, then is under 30 ℃ of temperature, rotating speed 200r/min at culture condition, fermentation 60h.
HPLC detects polydatin and Resveratrol content in fermented liquid, and obtaining polydatin, to be converted into trans-resveratrol transformation efficiency be 76.2%, transforms Resveratrol content in secondary fermentation liquid and reach 41.873 u g/mL, is unfermentable 7.89 times.
Claims (2)
1. polydatin is converted into a method for trans-resveratrol, it is characterized in that, comprises the following steps:
(1) endogenetic fungus L1 spore suspension preparation: in 10ml physiological saline, fully shake from Shang Tiao five rings, endogenetic fungus L1 PDA inclined-plane thalline;
Described endogenetic fungus L1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC NO.4558;
(2) substratum is prepared: fresh giant knotweed is crushed to 200 orders, in mass ratio giant knotweed 1.5wt%, dehydrated alcohol 2 wt%, CMC-Na 2.5 wt%, NH
4nO
30.2 wt%, Na
2hP0
40.06 wt%, CaCl
20.1 wt% preparation fermention medium, adjusting initial pH is 4.5-5.5;
(3) fermentation: first by the substratum in step (2) through 121 ℃, 30min moist heat sterilization; Again by endogenetic fungus L1 spore suspension, access in the substratum of initial pH 4.5-5.5, inoculum size is the 5-10% that is equivalent to culture volume, in 250ml triangular flask, the liquid amount of substratum is 30mL, under culture condition is temperature 26-30 ℃, rotating speed 200-220r/min, fermentation 36-60h.
2. polydatin according to claim 1 is converted into the method for trans-resveratrol, it is characterized in that, in step (3), the inoculum size of endogenetic fungus L1 spore suspension is to be equivalent to 9% of culture volume.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673681A (en) * | 2015-02-11 | 2015-06-03 | 青岛农业大学 | Inonotus obliquus QD04 and method for converting polygonum cuspidatum by same to produce resveratrol, triterpenoid saponin and polysaccharide |
| CN104774773A (en) * | 2015-04-30 | 2015-07-15 | 湖南农业大学 | Polygonum cuspidatum endogenous bacterial strain aspergillus fumigates J3 bacterial strain for converting polydatin |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101338327A (en) * | 2008-08-13 | 2009-01-07 | 长沙华诚生物科技有限公司 | Process for extracting resveratrol with purity higher than 98 0.000000rom giant knotweed |
| CN102199548A (en) * | 2011-05-18 | 2011-09-28 | 湖南农业大学 | Polygonum cuspidatum root endophytic strain G3 for efficiently transforming polydatin into resveratrol |
-
2013
- 2013-12-24 CN CN201310720687.1A patent/CN103695478A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101338327A (en) * | 2008-08-13 | 2009-01-07 | 长沙华诚生物科技有限公司 | Process for extracting resveratrol with purity higher than 98 0.000000rom giant knotweed |
| CN102199548A (en) * | 2011-05-18 | 2011-09-28 | 湖南农业大学 | Polygonum cuspidatum root endophytic strain G3 for efficiently transforming polydatin into resveratrol |
Non-Patent Citations (1)
| Title |
|---|
| 刘华金: "虎杖内生菌高效转化白藜芦醇苷的研究", 《中国优秀硕士学位论文全文数据库》, no. 12, 15 December 2012 (2012-12-15), pages 055 - 16 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673681A (en) * | 2015-02-11 | 2015-06-03 | 青岛农业大学 | Inonotus obliquus QD04 and method for converting polygonum cuspidatum by same to produce resveratrol, triterpenoid saponin and polysaccharide |
| CN104673681B (en) * | 2015-02-11 | 2017-12-22 | 青岛农业大学 | The method of one plant of Inonotus obliquus QD04 and its converting giant knotweed generation resveratrol, triterpenoid saponin and polysaccharide |
| CN104774773A (en) * | 2015-04-30 | 2015-07-15 | 湖南农业大学 | Polygonum cuspidatum endogenous bacterial strain aspergillus fumigates J3 bacterial strain for converting polydatin |
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Application publication date: 20140402 |