CN106167530A - A kind of combined-enzyme method extracts the method for ganoderan - Google Patents
A kind of combined-enzyme method extracts the method for ganoderan Download PDFInfo
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- CN106167530A CN106167530A CN201610532369.6A CN201610532369A CN106167530A CN 106167530 A CN106167530 A CN 106167530A CN 201610532369 A CN201610532369 A CN 201610532369A CN 106167530 A CN106167530 A CN 106167530A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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Abstract
The invention belongs to edible fungi deep process technology field, be specifically related to a kind of method that combined-enzyme method extracts ganoderan.The present invention uses combined-enzyme method to extract ganoderan, and compared with traditional hot water extraction method, crude polysaccharides yield and polyoses content can obtain bigger raising.Using the Ganoderma sporophore raw material of 40 mesh, Enzymatic Extraction ganoderma lucidum crude polysaccharide yield the most equally is 4.41%, and polyoses content is 64.10%；Traditional hot water extraction method's crude polysaccharides yield is 3.12%, and polyoses content is 55.81%.Crude polysaccharides yield improves 41.3%, and polyoses content improves 14.85%.
Description
Technical field
The invention belongs to edible fungi deep process technology field, be specifically related to a kind of combined-enzyme method and extract the side of ganoderan
Method.
Background technology
The health benefit of the mankind is increasingly paid attention to by edible fungi by the extensive of international community.Edible fungi polysaccharide is as edible
One of main active of bacterium, has become the study hotspot of World Health industry.Numerous studies show, edible fungi polysaccharide has
There are the multiple biological activitys such as immunomodulating, antitumor, antiviral, defying age, antioxidation, blood sugar lowering, blood fat and blood pressure.At present,
The state such as Asian countries headed by China and America and Europe is to multiple foods such as Ganoderma, Lentinus Edodes, Agaricus blazei Murrill, Grifola frondosa, Coprinus comatus
Having carried out more systematic research with granulose, wherein Ganoderma (Ganoderma lucidum) and ganoderan are unique with it
Health care become the focus of research.
Ganoderan is made up of, has the glucosan of helical form spatial configuration (tertiary structure) three strands of monosaccharide chains, its
Spatial configuration and DNA (deoxyribonucleic acid) (DNA), ribonucleic acid (RNA) are similar, are a kind of macromolecular compounds, and molecular weight is from thousand of
To hundreds of thousands.Ganoderan is present on the cell wall inwall of Ganoderma, can extract from Ganoderma spore powder or Ganoderma sporophore
Obtain.The monosaccharide composition predominantly glucose of ganoderan, secondly possibly together with arabinose, xylose, galactose, fucose, sweet
The monosaccharide such as dew sugar, rhamnose, but content is less.
Ganoderan can be dissolved in hot water, the ethanol insoluble in high concentration, can hence with this character employing decoction and alcohol sedimentation technique
To extract ganoderan.Traditional extracting method mainly has hot water extraction method, acid extraction method, alkali extraction method etc..Due to Ganoderma
Entity is made up of main components such as cellulose, hemicellulose, lignins, close structure, and active polysaccharide effective ingredient is attached to
Inside cell wall, the most traditional water extraction extraction time length, active component extraction ratio are low.Although acid extraction method and alkali extraction method
Polysaccharide extract rate can be improved, but to equipment requirements height, and the structure of active polysaccharide is easily destroyed, the ganoderan obtained produces
Product color is deep, impurity is many, is unfavorable for carrying out further research and development as functional food or medicine, there is also environmental pollution
Problem.The ultrasonic extraction and the microwave loss mechanisms that grow up subsequently are notable for the extraction effect of ganoderan.Carry with water
Method is compared, and has the clear superiorities such as extraction ratio is high, extraction time is short, but equipment needed thereby cost of investment is too big, the most still
Unrealized industrialized production.
Summary of the invention
In order to overcome the problem in the presence of prior art, it is an object of the invention to provide a kind of combined-enzyme method and extract spirit
The method of sesame polysaccharide.
To achieve these goals and other relevant purposes, the present invention adopts the following technical scheme that
A first aspect of the present invention, it is provided that a kind of combined-enzyme method extract ganoderan method, for use cellulase,
Pectase, wall breaking enzyme, protease are used for extracting ganoderan as compound enzyme.
Preferably, any one or more in papain, neutral protease, acid protease of described protease
Combination.
Described cellulase, pectase, wall breaking enzyme, papain, neutral protease, acid protease all can pass through city
Approach of selling obtains.
Preferably, described method comprises the steps:
(1) pulverize: carry out dry Ganoderma pulverizing acquisition Ganoderma powder；
(2) enzymolysis: gained Ganoderma powder in step (1) is added water according to certain solid-to-liquid ratio, stirs, regulation system pH
Value, adds described compound enzyme, and heated and stirred, carries out enzymolysis；After enzymolysis terminates, hydrolyzed solution boils enzyme denaturing；
(3) water carries for the first time: adjust temperature of reaction system, and water carries certain time, extracts after terminating centrifugal, it is thus achieved that for the first time
Supernatant and for the first time residue；
(4) water carries for the second time: by the residue for the first time of gained in step (3), adds water according to certain solid-to-liquid ratio, carries out second
Secondary water carries, and extracts after terminating centrifugal, obtains second time supernatant and second time residue；
(5) concentrate: merge supernatant and second time supernatant for the first time, concentrate and remove moisture removal, it is thus achieved that concentrated solution；
(6) precipitate with ethanol, filter, be dried: will in step (5) gained concentrated solution add ethanol, precipitation, filter, be dried obtain spirit
Sesame polysaccharide.
Preferably, in step (1), crusher for Chinese herbal medicine or super micron mill during pulverizing, is used to pulverize.
Preferably, in step (1), the mesh number of described Ganoderma powder is 10～2000 mesh.
Preferably, in step (1), in described compound enzyme, the addition of every kind of enzyme is the 0.5～5% of Ganoderma powder quality.
It is further preferred that the addition of every kind of enzyme is the 1～3% of Ganoderma powder quality in described compound enzyme.
It is further preferred that interpolation quality is Ganoderma powder quality the 1.0～5.0% of described protease.Described wall breaking enzyme
Interpolation quality is Ganoderma powder quality 2.0～5.0%.Described cellulase interpolation quality is Ganoderma powder quality 1.5～
5.0%.The addition of described pectase is the 0.5～2.0% of Ganoderma powder quality.
Preferably, in step (2), the scope of described solid-to-liquid ratio is: 1:10～1:30.
Preferably, in step (2), the scope of described pH value is: 3.0～7.0.
Preferably, in step (2), temperature range during described enzymolysis is 35～60 DEG C.The time range of described enzymolysis is:
1～4h.
Preferably, in step (2), after enzymolysis terminates, hydrolyzed solution boils enzyme denaturing 5～10min enzyme denaturing.
Preferably, in step (3), temperature range when described water carries is: 80～100 DEG C.Time model when described water carries
Enclose and be: 1～2h.
Preferably, in step (4), temperature range when described water carries is: 90～100 DEG C.Time model when described water carries
Enclose and be: 1～2h.
Preferably, in step (4), the scope of described solid-to-liquid ratio is: 1:10～1:30.
Preferably, in step (5), use during described concentration and be concentrated in vacuo.
Preferably, in step (6), add ethanol to final concentration of the 50～90% of ethanol.
Preferably, in step (6), when being dried, use vacuum drying or lyophilization.
Compared with prior art, there is advantages that
The present invention uses combined-enzyme method to extract ganoderan, and compared with traditional hot water extraction method, crude polysaccharides yield is with many
Sugar content can obtain bigger raising.Using the Ganoderma sporophore raw material of 40 mesh the most equally, Enzymatic Extraction ganoderma lucidum crude polysaccharide obtains
Rate is 4.41%, and polyoses content is 64.10%；Traditional hot water extraction method's crude polysaccharides yield is 3.12%, and polyoses content is
55.81%.Crude polysaccharides yield improves 41.3%, and polyoses content improves 14.85%.
Accompanying drawing explanation
Fig. 1: combined-enzyme method of the present invention extracts the process chart of ganoderan.
Detailed description of the invention
Before further describing the specific embodiment of the invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiments；It is also understood that the term used in the embodiment of the present invention is specific concrete in order to describe
Embodiment rather than in order to limit the scope of the invention.The test method of unreceipted actual conditions in the following example,
Generally according to normal condition, or according to the condition proposed by each manufacturer.
When embodiment provides numerical range, it should be appreciated that unless the present invention is otherwise noted, two ends of each numerical range
Between point and two end points, any one numerical value all can be selected for.Unless otherwise defined, in the present invention use all technology and
The same meaning that scientific terminology and those skilled in the art of the present technique are generally understood that.Except in embodiment use concrete grammar, equipment,
Outside material, according to those skilled in the art's grasp to prior art and the record of the present invention, it is also possible to use and this
Any method, equipment and the material of the prior art that the method described in inventive embodiments, equipment, material are similar or equivalent comes real
The existing present invention.
The present invention uses combined-enzyme method as shown in Figure 1 to extract ganoderan.Specific embodiment is as follows:
Embodiment 1
Weigh the Ganoderma sporophore powder being crushed to 10 mesh, add water according to solid-to-liquid ratio 1:30, stir, regulation system pH
Value is 3.0, on the basis of Ganoderma powder quality, adds acid protease 1.0%, wall breaking enzyme 2.5%, cellulase 5.0%, pectin
Enzyme 1.0%, heated and stirred at a temperature of 60 DEG C, enzymolysis and extraction 4h.After enzymolysis terminates, hydrolyzed solution boils 5min enzyme denaturing.Afterwards will
Reaction system is warming up to 100 DEG C, and water carries 2h for the first time, and extraction terminates rear 12000r/min and is centrifuged 15min, and supernatant is standby, residual
Slag adds water by initial feed solid-to-liquid ratio 1:30, and 100 DEG C carry out second time water and carry, and extraction terminates rear 12000r/min and is centrifuged 15min,
Merge twice supernatant.Utilize Rotary Evaporators by the 1/10 of supernatant concentration to original volume, in concentrated solution, add anhydrous second
Alcohol is to ethanol final concentration of 90%, and-4 DEG C stand overnight, and to precipitate Ganoderma macromolecular polysaccharide, filters, and solid lyophilization obtains
Ganoderan.Through analyzing, ganoderma lucidum crude polysaccharide yield is 3.34%, and polyoses content is 60.58%.
Embodiment 2
Weigh the Ganoderma sporophore powder being crushed to 2000 mesh, add water according to solid-to-liquid ratio 1:10, stir, regulation system
PH value is 7.0, on the basis of Ganoderma powder quality, adds neutral protease 3.0%, wall breaking enzyme 5.0%, cellulase 2.0%, really
Glue enzyme 0.5%, heated and stirred at a temperature of 55 DEG C, enzymolysis and extraction 1h.After enzymolysis terminates, hydrolyzed solution boils 5min enzyme denaturing.Afterwards
Reaction system is warming up to 80 DEG C, and water carries 2h for the first time, and extraction terminates rear 12000r/min and is centrifuged 15min, and supernatant is standby, residual
Slag adds water by initial feed solid-to-liquid ratio 1:10, and 90 DEG C carry out second time water and carry, and extraction terminates rear 12000r/min and is centrifuged 15min,
Merge twice supernatant.Utilize Rotary Evaporators by the 1/10 of supernatant concentration to original volume, in concentrated solution, add anhydrous second
Alcohol is to ethanol final concentration of 70%, and-4 DEG C stand overnight, and to precipitate Ganoderma macromolecular polysaccharide, filters, and solid lyophilization obtains
Ganoderan.Through analyzing, ganoderma lucidum crude polysaccharide yield is 8.34%, and polyoses content is 80.55%.
Embodiment 3
Weigh the Ganoderma sporophore powder being crushed to 40 mesh, add water according to solid-to-liquid ratio 1:20, stir, regulation system pH
Value is 5.0, on the basis of Ganoderma powder quality, adds papain 1.5%, wall breaking enzyme 2.0%, cellulase 1.5%, pectin
Enzyme 2.0%, heated and stirred at a temperature of 50 DEG C, enzymolysis and extraction 2h.After enzymolysis terminates, hydrolyzed solution boils 5min enzyme denaturing.Afterwards will
Reaction system is warming up to 100 DEG C, and water carries 1.5h for the first time, and extraction terminates rear 12000r/min and is centrifuged 15min, and supernatant is standby,
Residue adds water by initial feed solid-to-liquid ratio 1:15, and 100 DEG C carry out second time water and carry, and extraction terminates rear 12000r/min and is centrifuged
15min, merges twice supernatant.Utilize Rotary Evaporators by the 1/10 of supernatant concentration to original volume, add in concentrated solution
Dehydrated alcohol is to ethanol final concentration of 90%, and-4 DEG C stand overnight, and to precipitate Ganoderma macromolecular polysaccharide, filters, and solid freezing is done
Dry obtain ganoderan.Through analyzing, ganoderma lucidum crude polysaccharide yield is 4.41%, and polyoses content is 64.10%.
Embodiment 4
Weigh the Ganoderma sporophore powder being crushed to 500 mesh, add water according to solid-to-liquid ratio 1:16, stir, regulation system
PH value is 4.6, on the basis of Ganoderma powder quality, adds papain 5.0%, wall breaking enzyme 2.1%, cellulase 4.5%, really
Glue enzyme 1.0%, heated and stirred at a temperature of 35 DEG C, enzymolysis and extraction 4h.After enzymolysis terminates, hydrolyzed solution boils 5min enzyme denaturing.Afterwards
Reaction system is warming up to 90 DEG C, and water carries 1h for the first time, and extraction terminates rear 12000r/min and is centrifuged 15min, and supernatant is standby, residual
Slag adds water by initial feed solid-to-liquid ratio 1:20, and 100 DEG C carry out second time water and carry, and extraction terminates rear 12000r/min and is centrifuged 15min,
Merge twice supernatant.Utilize Rotary Evaporators by the 1/10 of supernatant concentration to original volume, in concentrated solution, add anhydrous second
Alcohol is to ethanol final concentration of 50%, and-4 DEG C stand overnight, and to precipitate Ganoderma macromolecular polysaccharide, filters, and solid lyophilization obtains
Ganoderan.Through analyzing, ganoderma lucidum crude polysaccharide yield is 6.16%, and polyoses content is 75.59%.
From above-described embodiment acquired results, the combined-enzyme method of the present invention is used to extract ganoderan, with traditional heat
Water extraction method is compared, and crude polysaccharides yield and polyoses content can obtain bigger raising.Such as, Ganoderma of same use 40 mesh is real
Body raw material, using the combined-enzyme method of the present invention to extract ganoderma lucidum crude polysaccharide yield is 4.41%, and polyoses content is 64.10%；And adopt
Being 3.12% by traditional hot water extraction method's crude polysaccharides yield, polyoses content is 55.81%.Therefore, compared to hot water extraction
Method, uses the combined-enzyme method of the present invention to extract ganoderan, and crude polysaccharides yield at least improves 41.3%, and polyoses content improves
14.85%.
The above, only presently preferred embodiments of the present invention, not any formal and substantial to present invention restriction,
It should be pointed out that, for those skilled in the art, on the premise of without departing from the inventive method, also can make
Some improvement and supplement, these improve and supplement also should be regarded as protection scope of the present invention.All those skilled in the art,
Without departing from the spirit and scope of the present invention, make when available disclosed above technology contents a little more
The equivalent variations moved, modify and develop, is the Equivalent embodiments of the present invention；Meanwhile, all substantial technological pair according to the present invention
The change of any equivalent variations that above-described embodiment is made, modify and develop, all still fall within the scope of technical scheme
In.
Claims (10)
1. the method that combined-enzyme method extracts ganoderan, for using cellulase, pectase, wall breaking enzyme, protease conduct
Compound enzyme is used for extracting ganoderan.
Method the most according to claim 1, it is characterised in that described protease selected from papain, neutral protease,
The combination of any one or more in acid protease.
Method the most according to claim 1, it is characterised in that described method comprises the steps:
(1) pulverize: carry out dry Ganoderma pulverizing acquisition Ganoderma powder；
(2) enzymolysis: gained Ganoderma powder in step (1) is added water according to certain solid-to-liquid ratio, stirs, regulation system pH value,
Add described compound enzyme, and heated and stirred, carry out enzymolysis；After enzymolysis terminates, hydrolyzed solution boils enzyme denaturing；
(3) water carries for the first time: adjust temperature of reaction system, and water carries certain time, extracts after terminating centrifugal, it is thus achieved that supernatant for the first time
Liquid and for the first time residue；
(4) water carries for the second time: by the residue for the first time of gained in step (3), adds water according to certain solid-to-liquid ratio, carries out second time water
Carry, extract after terminating centrifugal, obtain second time supernatant and second time residue；
(5) concentrate: merge supernatant and second time supernatant for the first time, concentrate and remove moisture removal, it is thus achieved that concentrated solution；
(6) precipitate with ethanol, filter, be dried: will in step (5) gained concentrated solution add ethanol, precipitation, filter, be dried obtain Ganoderma many
Sugar.
Method the most according to claim 3, it is characterised in that in step (1), uses crusher for Chinese herbal medicine or super during pulverizing
Atomizer is pulverized.
Method the most according to claim 3, it is characterised in that in step (1), the mesh number of described Ganoderma powder is 10～2000
Mesh.
Method the most according to claim 3, it is characterised in that in step (1), the addition of every kind of enzyme in described compound enzyme
For Ganoderma powder quality 0.5～5%.
Method the most according to claim 3, it is characterised in that in step (2), the scope of described solid-to-liquid ratio is: 1:10～
1:30。
Method the most according to claim 3, it is characterised in that in step (2), the scope of described pH value is: 3.0～7.0.
Method the most according to claim 3, it is characterised in that in step (2), temperature range during described enzymolysis be 35～
60℃；The time range of described enzymolysis is: 1～4h.
Method the most according to claim 3, it is characterised in that described method also include any one in following characteristics or
Multinomial:
A): in step (3) and step (4), temperature range when described water carries is: 80～100 DEG C；Time model when described water carries
Enclose and be: 1～2h；
B): in step (4), the scope of described solid-to-liquid ratio is: 1:10～1:30；
C): in step (5), during described concentration, employing is concentrated in vacuo；
D): in step (6), ethanol is added to final concentration of the 50～90% of ethanol；
E): in step (6), when being dried, vacuum drying or lyophilization are used.
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Cited By (9)
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| CN107267571A (en) * | 2017-07-15 | 2017-10-20 | 合肥市晶谷米业有限公司 | A kind of extracting method of rice bran polysaccharide |
| CN108719773A (en) * | 2018-05-31 | 2018-11-02 | 张顺兴 | A kind of glossy ganoderma powder and preparation method thereof |
| CN109694416A (en) * | 2019-01-03 | 2019-04-30 | 湖北省农业科学院农产品加工与核农技术研究所 | A kind of high-purity, highly dissoluble, high activity edible fungi polysaccharide preparation method |
| CN110922447A (en) * | 2019-12-31 | 2020-03-27 | 武夷山元生泰生物科技有限公司 | Method for extracting triterpenic acid and polysaccharide from ganoderma lucidum |
| CN111705093A (en) * | 2020-06-29 | 2020-09-25 | 天津科技大学 | Method for preparing polysaccharide by fermenting ganoderma lucidum fruiting bodies with green trichoderma |
| CN112675204A (en) * | 2020-12-07 | 2021-04-20 | 上饶农业技术创新研究院 | Extraction method and extraction reaction tank for ganoderma lucidum active substances |
| CN115152845A (en) * | 2021-07-29 | 2022-10-11 | 王天明 | Natural dairy product endogenous stabilizer and application thereof |
| CN116731220A (en) * | 2023-06-19 | 2023-09-12 | 临沂大学 | Preparation method of black currant phyllosphere polysaccharide |
| CN117801131A (en) * | 2023-12-29 | 2024-04-02 | 临沂信邦生物科技有限公司 | Extraction method of fungus polysaccharide |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107267571A (en) * | 2017-07-15 | 2017-10-20 | 合肥市晶谷米业有限公司 | A kind of extracting method of rice bran polysaccharide |
| CN108719773A (en) * | 2018-05-31 | 2018-11-02 | 张顺兴 | A kind of glossy ganoderma powder and preparation method thereof |
| CN109694416A (en) * | 2019-01-03 | 2019-04-30 | 湖北省农业科学院农产品加工与核农技术研究所 | A kind of high-purity, highly dissoluble, high activity edible fungi polysaccharide preparation method |
| CN110922447A (en) * | 2019-12-31 | 2020-03-27 | 武夷山元生泰生物科技有限公司 | Method for extracting triterpenic acid and polysaccharide from ganoderma lucidum |
| CN111705093A (en) * | 2020-06-29 | 2020-09-25 | 天津科技大学 | Method for preparing polysaccharide by fermenting ganoderma lucidum fruiting bodies with green trichoderma |
| CN112675204A (en) * | 2020-12-07 | 2021-04-20 | 上饶农业技术创新研究院 | Extraction method and extraction reaction tank for ganoderma lucidum active substances |
| CN115152845A (en) * | 2021-07-29 | 2022-10-11 | 王天明 | Natural dairy product endogenous stabilizer and application thereof |
| CN115152845B (en) * | 2021-07-29 | 2023-09-26 | 王天明 | Natural dairy product endogenous stabilizer and application thereof |
| CN116731220A (en) * | 2023-06-19 | 2023-09-12 | 临沂大学 | Preparation method of black currant phyllosphere polysaccharide |
| CN116731220B (en) * | 2023-06-19 | 2024-01-23 | 临沂大学 | Preparation method of black currant phyllosphere polysaccharide |
| CN117801131A (en) * | 2023-12-29 | 2024-04-02 | 临沂信邦生物科技有限公司 | Extraction method of fungus polysaccharide |
| CN117801131B (en) * | 2023-12-29 | 2024-08-13 | 临沂信邦生物科技有限公司 | Extraction method of fungus polysaccharide |
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