CN106832032A - A kind of method for extracting tremella polysaccharides and white fungus albumen - Google Patents
A kind of method for extracting tremella polysaccharides and white fungus albumen Download PDFInfo
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- CN106832032A CN106832032A CN201710048941.6A CN201710048941A CN106832032A CN 106832032 A CN106832032 A CN 106832032A CN 201710048941 A CN201710048941 A CN 201710048941A CN 106832032 A CN106832032 A CN 106832032A
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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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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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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Abstract
The present invention provides a kind of method for extracting tremella polysaccharides and white fungus albumen, including following preparation process：1）Choose, crush；2）Degreasing；3）Alkali carries；4）Separate；5）Extract tremella polysaccharides；6）The method for extracting white fungus albumen, this extraction tremella polysaccharides for providing of the invention and white fungus albumen, extracts easy, and extraction efficiency is high, can effectively improve the purity of extraction, the feature of material after enhancing extraction.
Description
Technical field
The present invention relates to technical field of bioengineering, especially a kind of method for extracting tremella polysaccharides and white fungus albumen.
Background technology
White fungus (Tremella fuciformls) is that a kind of economic worth is high, and nutritious medicine-food two-purpose bacterium has
Nourishing promote the production of body fluid, moistening lung nourishing the stomach the effect of.Tremella polysaccharides is the main active of white fungus, with raising body immunity, drop
Blood sugar, reducing blood lipid, anti-aging, antiulcer, antithrombus formation, anti-mutation etc. are acted on.At present, the conventional extraction of tremella polysaccharides
It is Hot water extraction, but this traditional extracting method has that recovery rate is low, takes, causes.Silver
Fungus polysaccharides are divided into exocellular polysaccharide and cell wall polysaccharide, and cell wall polysaccharide water is difficult to extract complete, even diluted acid or diluted alkaline extraction method,
Again because par-tial polysaccharide is easily hydrolyzed, the active structure of polysaccharide is destroyed, cause polysaccharide loss and recovery rate to reduce.
The existing technology extracted in white fungus, such as a kind of method for extracting tremella polysaccharides and white fungus albumen, invention is related to
And the extracting method of a kind of tremella polysaccharides and white fungus albumen, including：（1）The activation culture and fermented and cultured of white fungus bacterium；（2）Ultrafiltration
Film concentrates and crushes tremella spore；（3）The preparation of polyethylene glycol/inorganic salts double-aqueous phase system；（4）Polyethylene glycol/inorganic salts are double
Aqueous phase system is extracted；（5）Freeze-drying prepares tremella polysaccharides；（6）Polyethylene glycol/inorganic salts double-aqueous phase system second extraction, surpasses
Membrane filtration, freeze-drying prepares white fungus albumen.The invention can realize tremella spore production and the serialization of Polyose extraction, rule
Modelling, is easy to the industrialized production of tremella polysaccharides；The intracellular polyse of existing tremella spore, there is tremella spore again in the product for obtaining
The exocellular polysaccharide that liquid fermentation is produced, greatly improves the yield of tremella polysaccharides；And tremella polysaccharides and protein is obtained together
When separate, it is to avoid the drawbacks of existing sevag methods go complex operations and the machine solvent contamination of isolating protein, improve polysaccharide
Purity.
The method of this extraction tremella polysaccharides and white fungus albumen of the invention, extracts complex, and extraction time long, and carries
The tremella polysaccharides and white fungus albumen relative purity for taking are poor.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of raising Polyose extraction efficiency, and yield is high, improves and extracts
Purity, extracts the easy method for extracting tremella polysaccharides and white fungus albumen.
To solve above-mentioned existing technical problem, the present invention uses following scheme：One kind extracts tremella polysaccharides and white fungus egg
White method, including following preparation process：
1）Choose white fungus to be cleaned with clear water, microwave drying 15-23min is crushed to 100-150 mesh, obtains white fungus powder；
2）1.2-1.7 times of ethanol ether mixed liquor is added in white fungus powder, filter residue is filtered to take after backflow 1-3h；
3）2-3 times of clear water is added in filter residue, stirring, ultrasonic wave hydrotropy immersion adds the hydrogen of naoh concentration 1-2%
Aqueous solution of sodium oxide adjusts pH value to immersion after 10-11, and backflow extraction 2-3 times merges to obtain leaching liquor；
4）It is the ethanol of 95-98% that saturation degree is added in leaching liquor, and stirring, immersion obtains sediment；
5）Milipore filter concentration after the filter aid of deionized water and 0.01-0.02% is added in sediment, is dried, obtain tremella polysaccharides；
6）2-3 times of distilled water is added in tremella polysaccharides, chloroform and n-butanol is sequentially added, 20-35min is shaken, centrifugation takes
The liquid of water layer and solution layer intersection, milipore filter concentration, dries, and obtains white fungus albumen.
Preferably, in step 2 in ethanol ether mixed liquor ethanol and ether ratio:1-1.2：1, add ethanol ether
Temperature is heated to 52-61 DEG C during mixed liquor, makes white fungus effective degreasing in ethanol ether, is carried to improve extraction polysaccharide and albumen
For convenient.
Preferably, also needing to add complex enzyme when adding sodium hydrate aqueous solution to soak in step 3,1-2.3h is digested, after
Gone out enzyme 10-17min, and immersion is soaked into a point enzymolysis using complex enzyme, and cell membrane is destroyed by complex enzyme, is improved and is extracted polysaccharide and egg
White purity.
Preferably, complex enzyme is cellulase and pectase presses 1：The ratio of 3-4.6 is mixed, according to a certain percentage
Regulation uses enzyme, improves enzymolysis quality and enzymolysis efficiency.
Preferably, the composition and its weight portion of filter aid are in step 5：Montmorillonite 4-10 parts, calgon 0.1~
0.5 part, 4.2~6.3 parts of sodium carbonate, 5~8.4 parts of Ludox, 1.8~2.2 parts of aluminium hydroxide, 0.01-0.02 parts of Mayan indigo plant, sulphur
0.02-0.04 parts of cyanic acid guanidine, is fully adsorbed unwanted impurity by filter aid, improves DNA purity.
Preferably, using molecular cut off dense for the milipore filter of 3000~6000Dal when milipore filter is concentrated in step 5
Contracting, drying mode is freeze-drying, and the tissue of polysaccharide is effectively protected when drying, and improves the effect for extracting polysaccharide.
Preferably, tremella polysaccharides also needs to add the protease inhibitors of 0.02-0.04% after being dissolved in distilled water in step 6,
Prevent the PD when albumen is extracted, it is ensured that extract the purity of albumen.
Preferably, the addition of chloroform is the 20-30% of tremella polysaccharides solution in step 6, the addition of n-butanol is chlorine
The 25-38% of imitative solution.
Preferably, using molecular cut off dense for the milipore filter of 6000~10000Dal when milipore filter is concentrated in step 6
Contracting, drying mode is freeze-drying.
Beneficial effect：This extraction tremella polysaccharides and the method for white fungus albumen that the present invention is provided, extract easy, extract effect
Rate is high, can effectively improve the purity of extraction, the feature of material after enhancing extraction.Be probably superposition generate it is unexpected
Beneficiating ingredient or Synergistic interaction, Mayan is blue and guanidine thiocyanate is by aluminium hydroxide, the dissolving of calgon promotion polysaccharide,
So as to improve its yield, a kind of method for extracting tremella polysaccharides and white fungus albumen is provided present invention employs above-mentioned technical proposal,
The deficiencies in the prior art are compensate for, it is reasonable in design, it is easy to operate.
Specific embodiment
It is described in further detail with the following Examples：
Embodiment 1：
A kind of method for extracting tremella polysaccharides and white fungus albumen, including following preparation process：
1）Choose white fungus to be cleaned with clear water, microwave drying 15min is crushed to 100 mesh, obtains white fungus powder；
2）1.2 times of ethanol ether mixed liquor is added in white fungus powder, filter residue is filtered to take after backflow 1h；
3）2 times of clear water is added in filter residue, stirring, ultrasonic wave hydrotropy immersion adds the hydroxide of naoh concentration 1%
Sodium water solution adjusts pH value to immersion after 10, and backflow extraction 2 times merges to obtain leaching liquor；
4）The ethanol that saturation degree is 95% is added in leaching liquor, stirring, immersion obtains sediment；
5）Milipore filter concentration, dries after adding deionized water and 0.01% filter aid in sediment, obtains tremella polysaccharides；
6）2-3 times of distilled water is added in tremella polysaccharides, chloroform and n-butanol is sequentially added, 32min, centrifugation, water intaking is shaken
The liquid of layer and solution layer intersection, milipore filter concentration, dries, and obtains white fungus albumen.
In step 2 in ethanol ether mixed liquor ethanol and ether ratio 1.2：1, temperature when adding ethanol ether mixed liquor
52 DEG C are heated to, make white fungus effective degreasing in ethanol ether, provide convenient to improve extraction polysaccharide and albumen.
Also need to add complex enzyme when adding sodium hydrate aqueous solution to soak in step 3, digest 2.3h, after go out enzyme 17min, adopt
Immersion is soaked into a point enzymolysis with complex enzyme, cell membrane is destroyed by complex enzyme, improve the purity for extracting polysaccharide and albumen.
Complex enzyme is that cellulase and pectase press 1：4.6 ratio is mixed, and regulation according to a certain percentage uses enzyme,
Improve enzymolysis quality and enzymolysis efficiency.
The composition of filter aid and its preferred weight part are in step 5：10 parts of montmorillonite, 0.5 part of calgon, sodium carbonate
6.3 parts, 8.4 parts of Ludox, 2.2 parts of aluminium hydroxide, Mayan indigo plant 0.02 part, 0.04 part of guanidine thiocyanate will be not required to by filter aid
The impurity wanted fully is adsorbed, and improves DNA purity.
Molecular cut off is used to be concentrated for the milipore filter of 4000Dal when milipore filter is concentrated in step 5, drying mode is freezing
Dry, the tissue of polysaccharide is effectively protected when drying, improve the effect for extracting polysaccharide.
Tremella polysaccharides also needs to add 0.04% protease inhibitors after being dissolved in distilled water in step 6, prevents from extracting albumen
When PD, it is ensured that extract albumen purity.
The addition of chloroform is the 30% of tremella polysaccharides solution in step 6, and the addition of n-butanol is the 25- of chloroformic solution
38%。
Molecular cut off is used to be concentrated for the milipore filter of 8000Dal when milipore filter is concentrated in step 6, drying mode is freezing
Dry.
Embodiment 2：
A kind of method for extracting tremella polysaccharides and white fungus albumen, including following preparation process：
1）Choose white fungus to be cleaned with clear water, microwave drying 15min, microwave frequency is 3000MHz, is crushed to 100 mesh, obtains white fungus powder
End；
2）The ratio of the ethanol ether mixed liquor of 1.2 times of addition in white fungus powder, ethanol and ether is 1.2：1, add ethanol
Temperature is heated to 52 DEG C during ether mixed liquor, makes white fungus effective degreasing in ethanol ether, and polysaccharide and albumen are extracted to improve
It is convenient to provide, and filter residue is filtered to take after backflow 1h；
3）2 times of clear water is added in filter residue, stirring, ultrasonic wave hydrotropy immersion adds the hydroxide of naoh concentration 1%
Sodium water solution adjusts pH value to immersion after 11, also needs to add complex enzyme when adding sodium hydrate aqueous solution to soak, and digests 2.3h, after
Go out enzyme 17min, and wherein complex enzyme is that cellulase and pectase press 1：4.6 ratio is mixed, and is adjusted according to a certain percentage
Using enzyme, enzymolysis quality and enzymolysis efficiency are improved, immersion is soaked by a point enzymolysis using complex enzyme, cell is destroyed by complex enzyme
Wall, improves the purity for extracting polysaccharide and albumen, and micro sodium borohydride can be also added in order to prevent polysaccharide from degrading, and addition is
0.002%, backflow extraction 2 times merges to obtain leaching liquor；
4）The ethanol that saturation degree is 95% is added in leaching liquor, stirring, immersion obtains sediment；
5）Milipore filter concentration, dries after adding deionized water and 0.02% filter aid in sediment, obtains tremella polysaccharides, ultrafiltration
Film uses molecular cut off to be concentrated for the milipore filter of 5000Dal when concentrating, drying mode is freeze-drying, is effectively protected when drying
The tissue of polysaccharide is protected, the effect for extracting polysaccharide is improved；
6）2-3 times of distilled water is added in tremella polysaccharides, chloroform and n-butanol is sequentially added, the addition of chloroform is that white fungus is more
The 30% of sugar juice, the addition of n-butanol is the 25-38% of chloroformic solution, shakes 24min, and centrifugation, water intaking layer and solution layer are handed over
Liquid at boundary, milipore filter concentration, dries, and obtains white fungus albumen, and milipore filter uses molecular cut off surpassing for 8000Dal when concentrating
Filter membrane is concentrated, and drying mode is freeze-drying.
This extracted many-sided influence factor that center considers albumen and polysaccharide, using optimum environment to tremella polysaccharides
With the extraction of albumen, the concentration of extraction is effectively raised, and extraction rate is improve by techniques such as hydrotropy, degreasings, can had
Effect apply to enterprise production in.
Specific embodiment described herein is only to explanation for example of the invention.The technical field of the invention
Technical staff can be made various modifications or supplement to described specific embodiment or be substituted using similar mode, but
Without departing from of the invention or surmount scope defined in appended claims.
Claims (9)
1. a kind of method for extracting tremella polysaccharides and white fungus albumen, it is characterised in that including following preparation process：
1）Choose white fungus to be cleaned with clear water, microwave drying 15-23min is crushed to 100-150 mesh, obtains white fungus powder；
2）1.2-1.7 times of ethanol ether mixed liquor is added in white fungus powder, filter residue is filtered to take after backflow 1-3h；
3）2-3 times of clear water is added in filter residue, stirring, ultrasonic wave hydrotropy immersion adds the hydrogen of naoh concentration 1-2%
Aqueous solution of sodium oxide adjusts pH value to immersion after 10-11, and backflow extraction 2-3 times merges to obtain leaching liquor；
4）It is the ethanol of 95-98% that saturation degree is added in leaching liquor, and stirring, immersion obtains sediment；
5）Milipore filter concentration after the filter aid of deionized water and 0.01-0.02% is added in sediment, is dried, obtain tremella polysaccharides；
6）2-3 times of distilled water is added in tremella polysaccharides, chloroform and n-butanol is sequentially added, 20-35min is shaken, centrifugation takes
The liquid of water layer and solution layer intersection, milipore filter concentration, dries, and obtains white fungus albumen.
2. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
In 2 in ethanol ether mixed liquor ethanol and ether ratio:1-1.2：1, temperature is heated to 52- when adding ethanol ether mixed liquor
61℃。
3. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
Also need to add complex enzyme when adding sodium hydrate aqueous solution to soak in 3, digest 1-2.3h, after go out enzyme 10-17min.
4. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 4, it is characterised in that：It is described compound
Enzyme is that cellulase and pectase press 1：The ratio of 3-4.6 is mixed.
5. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
The composition and its weight portion of filter aid are in 5：Montmorillonite 4-10 parts, 0.1~0.5 part of calgon, sodium carbonate 4.2~6.3
Part, 5~8.4 parts of Ludox, 1.8~2.2 parts of aluminium hydroxide, 0.01-0.02 parts, guanidine thiocyanate 0.02-0.04 parts of Mayan indigo plant.
6. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
Molecular cut off is used to be concentrated for the milipore filter of 3000~6000Dal when milipore filter is concentrated in 5, drying mode is freeze-drying.
7. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
Tremella polysaccharides also needs to add the protease inhibitors of 0.02-0.04% after being dissolved in distilled water in 6.
8. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
The addition of chloroform is the 20-30% of tremella polysaccharides solution in 6, and the addition of n-butanol is the 25-38% of chloroformic solution.
9. a kind of method for extracting tremella polysaccharides and white fungus albumen according to claim 1, it is characterised in that：The step
Molecular cut off is used to be concentrated for the milipore filter of 6000~10000Dal when milipore filter is concentrated in 6, drying mode is freeze-drying.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383223A (en) * | 2017-06-22 | 2017-11-24 | 兰溪市哥特生物技术有限公司 | The extracting method of Auricularia polysaccharide |
| CN108359025A (en) * | 2018-04-13 | 2018-08-03 | 苏州尚格生物科技有限公司 | Tremella polysaccharides and the preparation method and application thereof |
| CN112251480A (en) * | 2020-10-16 | 2021-01-22 | 福建环海生物科技股份有限公司 | Production process of tremella oligosaccharide |
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| CN1580079A (en) * | 2004-05-21 | 2005-02-16 | 大连轻工业学院 | Method for extracting edible tree fungi polysaccharide |
| CN101157016A (en) * | 2007-07-25 | 2008-04-09 | 长兴仁恒精制膨润土有限公司 | A fluid-aid filter aid |
| CN105481993A (en) * | 2016-02-03 | 2016-04-13 | 赵志鑫 | Tremella polysaccharide extracting method |
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2017
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1580079A (en) * | 2004-05-21 | 2005-02-16 | 大连轻工业学院 | Method for extracting edible tree fungi polysaccharide |
| CN101157016A (en) * | 2007-07-25 | 2008-04-09 | 长兴仁恒精制膨润土有限公司 | A fluid-aid filter aid |
| CN105481993A (en) * | 2016-02-03 | 2016-04-13 | 赵志鑫 | Tremella polysaccharide extracting method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107383223A (en) * | 2017-06-22 | 2017-11-24 | 兰溪市哥特生物技术有限公司 | The extracting method of Auricularia polysaccharide |
| CN108359025A (en) * | 2018-04-13 | 2018-08-03 | 苏州尚格生物科技有限公司 | Tremella polysaccharides and the preparation method and application thereof |
| CN112251480A (en) * | 2020-10-16 | 2021-01-22 | 福建环海生物科技股份有限公司 | Production process of tremella oligosaccharide |
| CN112251480B (en) * | 2020-10-16 | 2022-03-08 | 福建环海生物科技股份有限公司 | Production process of tremella oligosaccharide |
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Application publication date: 20170613 |