CN1580271A - Method for producing flavone aglycone of ginkgo - Google Patents
Method for producing flavone aglycone of ginkgo Download PDFInfo
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- CN1580271A CN1580271A CN 03153489 CN03153489A CN1580271A CN 1580271 A CN1580271 A CN 1580271A CN 03153489 CN03153489 CN 03153489 CN 03153489 A CN03153489 A CN 03153489A CN 1580271 A CN1580271 A CN 1580271A
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Abstract
The invention publishes a kind of method to produce ginkgo flavones aglycon. First, biologically transfer ginkgo leaves by ferment and get ferment liquid. Second, primarily clean the liquid and brine down it to 1/2-1/5 of original volume. third, adsorb resin through macropore and wash it in distilled water to non color. Forth, strip in strippant and collect the flavones liquid. Fifth, condensing and get crude product and get high purity flavones aglycon by recrystallization. The raw material is dry or flesh leaves of ginkgo, which can be gotten easily and is environmental, and the manufactory is free from season. The content of flavones aglycon can measure up to 85% under the microbiology strain or biology enzyme action. This content is 5-20 times than the content in raw material. If adopt polymeric adsorbent in macropore and the technology of recrystallization, the content can increase 90% upwards.
Description
Technical field
The present invention relates to a kind of production method of ginkgo flavonoid glycoside, relate in particular to a kind of applying biological transformation technology and produce gingkgo genin type flavones, use the method for absorption and crystallization phases bonded technology purifying gingkgo genin type flavones, belong to biological technical field.
Background technology
Flavones has wide biological activity, comprises characteristics such as anti-oxidant, anti-ageing, antitumor, and the major physiological active substance is flavonoid compound in the ginkgo.Usually, contain abundant Flavonoid substances in the puerarin, its content is far above the content of flavones in the fruit; And the low grease content of blade also makes it become the high-quality source of flavones; On the other hand, in fruit, flavones mainly is to exist with the glucoside n-ary form n, and the flavones in blade mainly is to exist with the glycoside form.At present, domestic have utilize Ginkgo Leaf to extract flavones, basic nobody pays close attention to the configuration and the physiologically active height of its product.
Usually, the physiologically active of flavone glycoside is lower than glycosides million.Animal body shows also that to the assimilation effect of flavones flavones pathways metabolism in animal body is inequality, and glycosides million can directly be absorbed and enter in the animal blood; And most of flavone glycoside compound can not enter into blood by small bowel in human body, but lost by degraded of heterocycle fragmentation pattern and metabolism by microorganism in the enteric cavity, the small portion flavones is only arranged in the colon microorganism, as be distributed in the milk-acid bacteria in small intestine lower end and the large intestine and bifidus bacillus is secernent takes off under the effect of binding substances enzyme, hydrolysis generation glucoside unit reuptakes and enters blood, and promptly glycosides million has higher biological value than flavone glycoside.Therefore, produce glycosides million and have more obvious significance.
Bio-transformation, be to utilize animals and plants in vitro tissue, microorganism and enzyme etc. that xenobiontics is carried out structural modification and the biochemical reactions that obtains valuable product, its essence is that the enzyme that utilizes living things system itself to be produced carries out enzymic catalytic reaction to xenobiontics, advantages such as reaction preference is strong, reaction conditions is gentle, by product is few, non-environmental-pollution that it has.Since bio-transformation can carry out traditional organic synthesis can not or be difficult to the chemical reaction that carries out, bigger theory and actual application value are arranged.At present, adopt biological process very active to the research that functional ingredient transforms.Satoshi Tahara utilizes the Asperlliusflavus bacterium to transform flavones and obtains the poly-hydroxy flavones, has the more highly active product of more original glycoside, Lun Yuan (2002) utilizes that genistein is genistein in the fungal organism soybean transformation isoflavones, improves its cancer resistance.Aspect Potenlini, Takashi Kuramoto (1993) etc. transform Potenlini with mould (Cryptococus magnus MG-27), the single glucal acidic group of single acquisition glycyrrhetinic acid; Domestic, the red sudden strain of a muscle of fish (1999) etc. improve pol with mould and bacterium enzymatic conversion method Potenlini.Aspect the bio-transformation of other natural compounds, study also very active.But, have not yet to see research report about the ginkgolic flavone glycoside bio-transformation.
Summary of the invention
The objective of the invention is to propose a kind of production method of ginkgo flavonoid glycoside, utilize the content of conversion technology and absorption and crystallization phases bonded purification technique raising glucoside type ginkgolic flavone glycoside, thus the content of flavones in the physiologically active of raising ginkgolic flavone glycoside and the finished product.
A kind of production method of ginkgo flavonoid glycoside comprises:
At first, Ginkgo Leaf is carried out bio-transformation by fermentation, obtain fermented liquid;
Then, after the fermented liquid rough purification, be evaporated to the 1/2-1/5 of original volume, colourless by being washed with distilled water to washings after the polymeric adsorbent absorption, use the strippant desorb again, collect the stripping liquid that merging contains flavones, separate out crude product after concentrating, last recrystallization obtains highly purified glycosides million product.
Described Ginkgo Leaf is carried out bio-transformation by fermentation, can handle aquatic foods or dry Folium Ginkgo with the production by biological enzyme, also can be with prozyme effect Folium Ginkgo extract.
Describedly handle aquatic foods or dry Folium Ginkgo with the production by biological enzyme, its yeasting temperature is 25-40 ℃, and fermented liquid pH value is 4.5-6.5, the time be 5 hours-to 240 hours.
Described with prozyme effect Folium Ginkgo extract, 45-50 ℃ of its invert point, fermented liquid pH value is 4.5-6.5; Enzyme concentration 2-3 restrains/1 kilogram of ginkgo substrate; Transformation time is 60-90 minute;
Described polymeric adsorbent is a macroporous adsorbent resin.
The bio-transformation of described ginkgo flavonoid glycoside and purification purification process, carry out resulting fermentating liquid acidification is carried out acidification after also being included in the Ginkgo Leaf bio-transformation, with fermentating liquid acidification to pH value is 0.5-4.0, water bath reflux temperature 75-85 ℃, and acidificatoin time 1-2 hour.
The present invention does with ginkgo or bright leaf is that major ingredient is produced the glycosides million product, and raw material is gathered conveniently, and resource is not had destruction, realizes Sustainable utilization of resources, is not subject to seasonal restrictions; The present invention adopts the conversion technology of flavone glycoside unit in microorganism strains or the biological enzyme effect raw material to improve glycosides million content in the product, reach more than 85%, for the 5-20 of glycosides million content in the raw material doubly; The present invention can reach more than 90% with the content of macroporous adsorbent resin in conjunction with flavones in the recrystallization technology raising product.
Embodiment
Flavones mainly is to exist with the glucoside form in the Ginkgo Leaf, after flavone glycoside enters in the animal body at first metabolism be glycosides million, enter blood and bring into play its physiological function thereby could further be absorbed.But, in this process most of flavones all by enteric cavity in microorganism lose by degraded of heterocycle fragmentation pattern and metabolism.The present invention adopts the biotechnology means directly ginko leaves flavone to be converted into glycosides million; Adopt macroporous adsorbent resin directly from transformation system, to adsorb ginkgo flavonoid glycoside, can improve the content of ginkgo flavonoid glycoside product, through after the strippant desorb, it is ginkgolic flavone glycoside more than 90% that recrystallization can obtain purity, and wherein the content of glycosides million can reach more than 85%.
Ginkgo Leaf is carried out bio-transformation, can handle aquatic foods or dry Folium Ginkgo with the production by biological enzyme, embodiment 2 adopts this method that Ginkgo Leaf is carried out bio-transformation, and the yeasting temperature is 25-40 ℃, and fermented liquid pH value is 4.5-6.0, the time be 5 hours-to 240 hours;
Also can be with prozyme effect Folium Ginkgo extract, embodiment 1 adopts this method, and 45-50 ℃ of its invert point, pH value are 4.5-6.5, and enzyme concentration 2-3 restrains/1 kilogram of ginkgo substrate, and transformation time is 60-90 minute.
Also comprise fermented liquid is carried out acidification: fermentating liquid acidification to pH value is 0.5-4.0, water bath reflux temperature 75-85 ℃, acidificatoin time 1-2 hour.
The step of glycosides million purifying comprises, with the fermented liquid that before obtained after rough purification, be evaporated to the 1/2-1/5 of original volume, by the inertia macroporous adsorbent resin, it is colourless to be washed with distilled water to washings after the absorption, uses the strippant desorb then, collects to merge the stripping liquid that contains flavones, separate out crude product after concentrating, last recrystallization obtains highly purified glycosides million.
Method below in conjunction with embodiment 1 and embodiment 2 detailed description bio-transformations of the present invention and purification purifying.
Embodiment 1:
Get 100 gram ginko leaves flavone extracts, be scattered in 1000 ml waters, and be preheated to 40 ℃; Take by weighing enzyme 0.6 gram, use the 5ml dissolved in distilled water, join in the flavones dispersion liquid, 40 ℃ of constant temperature mixed 120 minutes; After the filtration, fermented liquid is evaporated to 1/2 of original volume; Concentrated solution carries out acidification after adding methyl alcohol (or ethanol) extraction, neutralization then, rotary evaporation neutralizer.
Then the concentrated solution that obtains is adsorbed by the inertia macroporous adsorptive resins, it is colourless substantially to be washed with distilled water to washings again, use the acetone stripping liquid then, collect and the merging stripping liquid, separate out the glycosides million crude product after concentrating, after acetone recrystallization obtains highly purified ginkgo flavonoid glycoside product.
Embodiment 2:
Get the bright leaf of 10 gram ginkgoes, after grinding, join in the microorganism strains nutrient solution that produces enzyme, temperature is 30 ℃, and the pH value is 5.0, and the time is fermentation altogether in 120 hours.Behind the filtering fermentation liquor, be evaporated to 1/5 of original volume; Concentrated solution adds methyl alcohol (or ethanol) and extracted 12-15 hour, carry out acidification, then and acidizing fluid, after being rotated the evaporation neutralizer, concentrated solution directly adsorbs by the inertia macroporous adsorptive resins, and it is colourless substantially to be washed with distilled water to washings again, use the acetone desorb then, collect and the merging stripping liquid, separate out the glycosides million crude product after concentrating, after acetone recrystallization obtains the ginkgo flavonoid glycoside product.
It should be noted last that: above embodiment is the unrestricted technical scheme of the present invention in order to explanation only, although the present invention is had been described in detail with reference to the foregoing description, those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not breaking away from any modification or partial replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (6)
1, a kind of production method of ginkgo flavonoid glycoside is characterized in that, comprising:
At first, Ginkgo Leaf is carried out bio-transformation by fermentation, obtain fermented liquid;
Then, after the fermented liquid rough purification, be evaporated to the 1/2-1/5 of original volume, colourless by polymeric adsorbent absorption after scouring to washings, use the strippant desorb again, collect the stripping liquid that merging contains flavones, separate out crude product after concentrating, last recrystallization obtains highly purified glycosides million product.
2, according to the production method of the described ginkgo flavonoid glycoside of claim 1, it is characterized in that, described Ginkgo Leaf is carried out bio-transformation by fermentation, can handle aquatic foods or dry Folium Ginkgo with the production by biological enzyme, also can be with prozyme effect Folium Ginkgo extract.
According to the production method of the described ginkgo flavonoid glycoside of claim 2, it is characterized in that 3, describedly handle aquatic foods or dry Folium Ginkgo with the production by biological enzyme, its yeasting temperature is 25-40 ℃, fermented liquid pH value is 4.5-6.0, the time be 5 hours-to 240 hours.
4, according to the production method of the described ginkgo flavonoid glycoside of claim 2, it is characterized in that, described with prozyme effect Folium Ginkgo extract, 45-50 ℃ of its invert point, fermented liquid pH value is 4.5-6.5; Enzyme concentration 2-3 restrains/1 kilogram of ginkgo substrate; Transformation time is 60-90 minute.
According to the production method of the described ginkgo flavonoid glycoside of claim 1, it is characterized in that 5, described polymeric adsorbent is a macroporous adsorbent resin.
6, according to the production method of the described ginkgo flavonoid glycoside of claim 1, it is characterized in that, carry out resulting fermentating liquid acidification is carried out acidification after also being included in the Ginkgo Leaf bio-transformation, with fermentating liquid acidification to pH value is 0.5-4.0, water bath reflux temperature 75-85 ℃, acidificatoin time 1-2 hour.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 03153489 CN1580271A (en) | 2003-08-14 | 2003-08-14 | Method for producing flavone aglycone of ginkgo |
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| CN 03153489 CN1580271A (en) | 2003-08-14 | 2003-08-14 | Method for producing flavone aglycone of ginkgo |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101100683B (en) * | 2006-09-08 | 2010-05-12 | 颜廷和 | Glucoside type flavone biological transformation and purification technique |
| CN101768613A (en) * | 2010-03-09 | 2010-07-07 | 中国农业大学 | Preparation method of flavone genin of seabuckthorn leaves |
-
2003
- 2003-08-14 CN CN 03153489 patent/CN1580271A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101100683B (en) * | 2006-09-08 | 2010-05-12 | 颜廷和 | Glucoside type flavone biological transformation and purification technique |
| CN101768613A (en) * | 2010-03-09 | 2010-07-07 | 中国农业大学 | Preparation method of flavone genin of seabuckthorn leaves |
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