CN108774638A - A method of improving polyporus rhinoceros cooke acid ganoderma lucidum polysaccharide - Google Patents
A method of improving polyporus rhinoceros cooke acid ganoderma lucidum polysaccharide Download PDFInfo
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- CN108774638A CN108774638A CN201810386559.0A CN201810386559A CN108774638A CN 108774638 A CN108774638 A CN 108774638A CN 201810386559 A CN201810386559 A CN 201810386559A CN 108774638 A CN108774638 A CN 108774638A
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- polyporus rhinoceros
- rhinoceros cooke
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- 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
- C12P33/00—Preparation of steroids
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Abstract
The invention discloses a kind of methods improving polyporus rhinoceros cooke acid ganoderma lucidum polysaccharide, belong to research and improve active material method field.The present invention is using polyporus rhinoceros cooke biomass and active material yield as standard, the yield that different orange peel water extracts are used for improving biomass and active material is added during its liquid fermentation, selects best orange peel and best orange peel concentration and best orange peel combined concentration.The present invention can make triterpene substance in the zymotic fluid of polyporus rhinoceros cooke improve 22.65%~34.55%, polyporus rhinoceros cooke acid content improves at least 1.13 times, and ganoderma polyoses content improves 55.2~85.9% by adding citrus fruits severe edema due to hypofunction of the spleen extract.
Description
Technical field
The present invention relates to a kind of methods improving polyporus rhinoceros cooke acid ganoderma lucidum polysaccharide, belong to research and improve active material method neck
Domain.
Background technology
Polyporus rhinoceros cooke Lignosus rhinoceros (Cooke) Ryvarden is a kind of large-scale food originating in torrid areas
Medicinal fungi, Polyporaceae have 400 years history away from the present, and legend is dripped for tigress milk gives birth in ground.It is distributed mainly on China
Some countries and regions of Hainan Province, Malaysia, Philippine, Thailand, Indonesia and East Africa, expensive, sclerotium is it
Medicinal effects, for treating liver cancer, chronic hepatitis, stomach trouble etc..It has been investigated that at least containing 2 kinds or more parents in polyporus rhinoceros cooke
Lipid steroidal or ter penoids；Water-soluble and very high, the high dietary fiber content of Water insoluble dietary fiber content in its sclerotium
Physical chemistry functional characteristic become reconciled etc., can be used as the dispensing of multiduty low calories food, and sclerotium has more as dietary fiber
A technical advantage, such as natural origin is less as the edible and medical fungi type of active component using sclerotium, can make full use of this
Characteristic reinforces product development, meets the health care market demand；Sclerotium water extract has antitumor and immunoregulation effect, but specific ingredient
There are no clear.In addition, mushroom scholar thinks that obliqua polysaccharide is the fungi polysaccharide for having development prospect, sclerotium polysaccharide can stimulate
The proliferation of the certain immunocytes of human body.In recent years, polyporus rhinoceros cooke wild resource excessively excavation is petered out, artificial cultivation and liquid
Fermentation is the effective way solved the problems, such as, since artificial cultivation takes time and effort consumptive material, so liquid fermentation will become most effective
Method.
Polyporus rhinoceros cooke exocellular polysaccharide and polyporus rhinoceros cooke acid are its 2 kinds of important metabolic activity products, but itself sclerotium
In content it is very low.
Invention content
The problem of in order to improve active material low output in polyporus rhinoceros cooke sclerotium, the present invention provides a kind of raising tiger breast spirits
The method of sesame acid ganoderma lucidum polysaccharide is to utilize the water of orange peel using the yield of biomass, exocellular polysaccharide and polyporus rhinoceros cooke acid as index
Extract is fermented, and optimal conditions of fermentation is sought, to improve the work of polyporus rhinoceros cooke fermentation synthesis exocellular polysaccharide and polyporus rhinoceros cooke acid
Industryization provides reference.
The object of the present invention is to provide a kind of methods improving polyporus rhinoceros cooke acid function ingredient yield, and the method is to tiger
The pericarp extract of citrus fruits is added in newborn spirit fermentation process.
In one embodiment of the invention, the citrus fruits include citrus, orange, shaddock, lemon.
In one embodiment of the invention, the pericarp extract is to crush 1~25g pericarps, addition 100~
150mL water, in the Fruit peel extract that fluidized state water-bath 30~60min times obtain.
In one embodiment of the invention, the method is based on polyporus rhinoceros cooke fluid nutrient medium, to culture medium
The middle Fruit peel extract that citrus fruits are added.
In one embodiment of the invention, the functional component includes polyporus rhinoceros cooke acid and/or exocellular polysaccharide.
In one embodiment of the invention, the pericarp extract of the citrus fruits is orange peel water extract, is pressed
It is prepared by following steps：The orange peel after 10g crushing, sieving is weighed, adds 50mL water in fluidized state water-bath 30min, is collected by filtration
Extracting solution.Extraction 1 time is repeated to be settled to 200mL after filtrate merges, obtain the orange peel water extract of a concentration of 50g/L.
In one embodiment of the invention, the various concentration pomelo peel is a concentration of of addition pomelo peel water extract
0-25g/L。
In one embodiment of the invention, the water, which carries, is carried out under 80 DEG C of water-baths.
In one embodiment of the invention, it is to extract 3 times repeatedly that the water, which carries, and extracting solution merges.
The present invention also provides the methods in terms of preparing the product containing polyporus rhinoceros cooke acid or polyporus rhinoceros cooke exocellular polysaccharide
Using.
Advantageous effect：The present invention can make triterpene in the zymotic fluid of polyporus rhinoceros cooke by adding citrus fruits severe edema due to hypofunction of the spleen extract
Substance improves 22.65%~34.55%, and polyporus rhinoceros cooke acid content improves at least 1.13 times, and ganoderma polyoses content improves 55.2
~85.9%.
Description of the drawings
Fig. 1 is influence of the different orange peels to polyporus rhinoceros cooke biomass and active material yield；
Specific implementation mode
Polyporus rhinoceros cooke biomass calculates：Zymotic fluid 50mL is taken to centrifuge, distilled water is washed 3 times, and drying to constant weight by precipitation；Institute
It is to centrifuge 10min at 10000rpm to state centrifugation；The washing is to remove the substance for being dissolved in water, leaves behind mycelium；It is described
Drying, is that mycelium is put into 60 DEG C of baking ovens；The constant weight is drying thalline mass conservation.
The heavy sugar of ethyl alcohol is centrifuging and taking supernatant, the heavy sugar of ethyl alcohol, filtering, obtains polysaccharide.Supernatant afterwards, i.e. extracting solution are added, and mix
Even, standing, obtains sediment；The ethyl alcohol sink sugar be 70-80% with volume fraction ethyl alcohol；The heavy sugar of the ethyl alcohol is to take centrifugation
The absolute ethyl alcohol of 3-4 times of volume is added in supernatant afterwards, i.e. extracting solution, and mixing, standing obtain sediment；It is described centrifugation be
10min is centrifuged under 8000rpm.
Polyporus rhinoceros cooke acid is to take the mycelium 1g that crushed, according to solid-liquid ratio 1:85% ethyl alcohol is added successively 80 in 40 amount
40min is extracted at DEG C 3 times, 3 extracting solutions are merged into concentration.Obtained extract 30mL ethyl acetate and 30mL water extraction
It takes 2 times, 2 extracting solutions is merged into concentration.It is redissolved with methanol, is settled to 10mL, surveyed at 254nm with ultraviolet specrophotometer
Its absorbance.Ganoderic acid A standard items are dissolved with methanol, standard curve is obtained under same measuring condition.The solid-liquid ratio is bacterium
The volume ratio of filament quality and Extraction solvent；85% ethyl alcohol is that the volume fraction of ethyl alcohol is 85%；The concentration is 50 DEG C
Temperature carries out in the case of vacuum distillation；The concentration is 45 DEG C of temperature, is carried out in the case of vacuum distillation；The extraction is by second
Acetoacetic ester and water dissolution condensed matter, remove water layer；The extraction is to extract 2 times repeatedly, and extract liquor merges；The redissolution is
Condensed matter is dissolved with methanol.
Exocellular polysaccharide measures：The making (phend-sulphuric acid) of standard curve：Standard curve is done with Glucose standards；With Portugal
The absorbance of a concentration of abscissa of grape sugar, each pipe is that ordinate draws standard curve, and linear regression equation：Y=
13.627x+0.0274 R2=0.9972.
Polyporus rhinoceros cooke acidity test：With ganoderic acid A standard items (purity>98%) standard curve is done, in the item that wavelength is 254nm
Ultraviolet absorption value is detected under part.With a concentration of abscissas of ganoderic acid A, light absorption value is ordinate, calibration curve equation：Y=
16.001x-0.0148 R2=0.9991.Polyporus rhinoceros cooke acid yield=polyporus rhinoceros cooke acid content × biomass.The polyporus rhinoceros cooke
Acid content is the mycelium polyporus rhinoceros cooke acid content of unit mass, and unit is mg/g.
Liquid Culture based formulas (based on g/100mL)：KH2PO40.15, sucrose 2, yeast powder 0.5.
Embodiment 1：It is prepared by orange peel water extract
10g orange peels (having crushed, be sieved) are weighed, adds 50mL water in fluidized state water-bath 30min, extraction is collected by filtration
Liquid.Extraction 1 time is repeated to be settled to 200mL after filtrate merges, obtain a concentration of 50g/L.
Embodiment 2：Add different citrus fruits severe edema due to hypofunction of the spleen extracts
Different citrus fruits severe edema due to hypofunction of the spleen extracts are prepared in the way of embodiment 1 respectively, by orange peel, pomelo peel, orange
Skin and lemon peel are added with the additive amount of final concentration 5g/L in liquid fermentation liquid culture medium respectively, study different orange peel water extracts
Influence to polyporus rhinoceros cooke liquid fermentation, as a result such as Fig. 1.
Biomass is carried in the case where same dose (5g/L) orange peel is added with any citrus severe edema due to hypofunction of the spleen is not added
The control experiment of object is compared, and lemon peel and pomelo peel have an inhibiting effect to the growth of polyporus rhinoceros cooke, the inhibition of lemon peel it is obvious that
Biomass decline it is very much, the growing state of this and Antrodia camphorata on the contrary, may lemon to the growth situations of different edible mushrooms not
Together；Pomelo peel, which has the growth of polyporus rhinoceros cooke, a little to be inhibited.Other two kinds of orange peels are to the growth of polyporus rhinoceros cooke almost without shadow
Ring, it is weaker to the mycelial stimulation of polyporus rhinoceros cooke, and orange peel to the growth-promoting effect of Antrodia camphorata it is obvious that fragrant citrus pericarp
Also very weak.The biomass of polyporus rhinoceros cooke control group is 24.45g/L, and lemon peel, pomelo peel, orange peel and the orange severe edema due to hypofunction of the spleen is added and carries
The biomass of object is respectively 19.69,21.40,24.54 and 24.49g/L (Fig. 1).
Embodiment 3：Influence of the orange peel water extract to polyporus rhinoceros cooke polysaccharide
For exocellular polysaccharide, control experiment group yield is 3.19g/L, in addition to fragrant citrus pericarp water extract is to the yield of exocellular polysaccharide
There are facilitation, other orange peel water extracts that can all inhibit the synthesis (table 1) of polysaccharide.
Influence of the different orange peels of table 1 to polyporus rhinoceros cooke polysaccharide
For polyporus rhinoceros cooke acid, 4 kinds of orange peel water extracts have certain promotion to the synthesis of polyporus rhinoceros cooke acid, wherein
The water extract of fragrant citrus pericarp and lemon peel is weaker to the stimulation of polyporus rhinoceros cooke acid, and orange peel and pomelo peel are compared with other citrus prickles
Sharp effect is stronger, and control experiment group polyporus rhinoceros cooke acid content is 5.21mg/g, adds the fermentation of orange peel and pomelo peel water extract
Content is respectively 6.39mg/g and 7.01mg/g in liquid, and 22.65% and 34.55% has been respectively increased, and shows both orange peels
Water extract is advantageous (Fig. 1) to the synthesis of triterpene substance.
Embodiment 4：Influence of the various concentration pomelo peel water extract to polyporus rhinoceros cooke acid
The pomelo peel water extract of different final concentrations is added into fermentation medium respectively in the way of embodiment 3, for
Biomass adds in the zymotic fluid of various concentration pomelo peel water extract and is below control experiment, illustrates within the scope of test dose,
Pomelo peel water extract can inhibit the mycelial growth of polyporus rhinoceros cooke, and concentration is higher, and polyporus rhinoceros cooke biomass is lower.For brave breast spirit
Sesame acid, content increase (table 2) with the raising of pomelo peel concentration.
Influence of the 2 various concentration pomelo peel of table to polyporus rhinoceros cooke
Embodiment 5：Influence of the various concentration fragrant citrus pericarp water extract to polyporus rhinoceros cooke polysaccharide
The fragrant citrus pericarp water extract of different final concentrations is added into fermentation medium respectively in the way of embodiment 3, for
Biomass, adding different fragrant citrus pericarp water extracts influences less the mycelial growth of polyporus rhinoceros cooke.For exocellular polysaccharide, fragrant citrus pericarp
Concentration between 5-15g/L when, polyoses content is almost unchanged, and when more than 15g/L, and polyporus rhinoceros cooke polysaccharide is increased with concentration
Content is lower, illustrates that the fragrant citrus pericarp of high concentration can inhibit the synthesis (table 3) of exocellular polysaccharide.
Influence of the 3 various concentration fragrant citrus pericarp of table to polyporus rhinoceros cooke
Comparative example 1：Influences of the Vc to polyporus rhinoceros cooke
Orange peel water extract is prepared in the way of embodiment 1, measures the Vc contents wherein contained, and measurement obtains orange peel
Vitamin C content is 28.5mg/L in water extract.
Into zymotic fluid, the vitamin C of addition and the same final concentration of orange peel water extract is not (to add orange peel water extract
Or ascorbic culture medium is as a control group), it cultivated, fermented under the same conditions.Side is carried out to the product in zymotic fluid
Key, the results show that it is little with check experiment difference, so eliminating the influence (table 4) that vitamin C ferments to polyporus rhinoceros cooke.
The influence that 4 Vc of table ferments to polyporus rhinoceros cooke
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not limited to the present invention, any to be familiar with this skill
The people of art can do various change and modification, therefore the protection model of the present invention without departing from the spirit and scope of the present invention
Enclosing be subject to what claims were defined.
Claims (9)
1. a kind of method improving polyporus rhinoceros cooke acid function ingredient yield, which is characterized in that added into the clever fermentation process of brave breast
The pericarp extract of citrus fruits.
2. according to the method described in claim 1, it is characterized in that, the functional component includes that polyporus rhinoceros cooke is sour and/or extracellular
Polysaccharide.
3. method according to claim 1 or 2, which is characterized in that the citrus fruits include citrus, orange, shaddock, lemon
Lemon.
4. according to any method of claims 1 to 3, which is characterized in that pericarp extract is to crush 1~25g pericarps,
100~150mL water is added, in the Fruit peel extract that fluidized state water-bath 30~60min times obtain.
5. method according to any one of claims 1 to 4, which is characterized in that based on polyporus rhinoceros cooke fluid nutrient medium, to
The Fruit peel extract of citrus fruits is added in culture medium.
6. according to the method described in claim 5, it is characterized in that, the Fruit peel extract of the citrus fruits is the citrus severe edema due to hypofunction of the spleen
Extract is prepared as follows：Will crush, sieving after orange peel in fluidized state 20~40min of water-bath, extraction is collected by filtration
Liquid, repeats extraction 1~3 time, and merging filtrate closes.
7. according to the method described in claim 5, it is characterized in that, the Fruit peel extract of the citrus fruits is the shaddock severe edema due to hypofunction of the spleen
Extract, a concentration of 0-25g/L.
8. a kind of method improving polyporus rhinoceros cooke acid biomass, which is characterized in that add Citrus into the clever fermentation process of brave breast
The pericarp extract of fruit；The citrus fruits include citrus, orange, shaddock, lemon.
9. any the method for claim 1~8 is in terms of preparing the product containing polyporus rhinoceros cooke acid or polyporus rhinoceros cooke exocellular polysaccharide
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109668851A (en) * | 2019-03-04 | 2019-04-23 | 安徽协和成制药有限公司 | The quality standard detecting method of monarch drug in a prescription glossy ganoderma extract in a kind of pearl ganoderma lucidum slice |
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| WO2008068602A2 (en) * | 2006-12-08 | 2008-06-12 | Atehortua Garces Lucia | A method to generate fungal biomass from a culture of differentiated mycelium |
| CN101492645A (en) * | 2008-01-21 | 2009-07-29 | 湖北工业大学 | Glossy ganoderma cell culture method for accelerating biosynthesis of ganoderic acid and ganoderma iucidum polysaccharide |
| CN105838620A (en) * | 2016-03-28 | 2016-08-10 | 北京七巧时代科技有限公司 | Ganoderma lucidum culture medium and ganoderma lucidum culture method |
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2018
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| WO2008068602A2 (en) * | 2006-12-08 | 2008-06-12 | Atehortua Garces Lucia | A method to generate fungal biomass from a culture of differentiated mycelium |
| CN101492645A (en) * | 2008-01-21 | 2009-07-29 | 湖北工业大学 | Glossy ganoderma cell culture method for accelerating biosynthesis of ganoderic acid and ganoderma iucidum polysaccharide |
| CN105838620A (en) * | 2016-03-28 | 2016-08-10 | 北京七巧时代科技有限公司 | Ganoderma lucidum culture medium and ganoderma lucidum culture method |
Non-Patent Citations (2)
| Title |
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| FAN-CHIANG YANG等: "Medium modification to enhance the formation of bioactive metabolites in shake flask cultures of Antrodia cinnamomea by adding citrus peel extract", 《BIOPROCESS BIOSYST ENG》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109668851A (en) * | 2019-03-04 | 2019-04-23 | 安徽协和成制药有限公司 | The quality standard detecting method of monarch drug in a prescription glossy ganoderma extract in a kind of pearl ganoderma lucidum slice |
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