CN113583145A - Method for extracting polysaccharide based on tremella spore fermentation liquor - Google Patents
Method for extracting polysaccharide based on tremella spore fermentation liquor Download PDFInfo
- Publication number
- CN113583145A CN113583145A CN202110721695.2A CN202110721695A CN113583145A CN 113583145 A CN113583145 A CN 113583145A CN 202110721695 A CN202110721695 A CN 202110721695A CN 113583145 A CN113583145 A CN 113583145A
- Authority
- CN
- China
- Prior art keywords
- steps
- tremella
- following
- extracting
- polysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a method for extracting polysaccharide based on tremella spore fermentation liquor, belonging to the technical field of preparation and application of polysaccharide, and the method for extracting polysaccharide based on tremella spore fermentation liquor comprises the following steps: step one, raw material preparation, filtration of tremella spore fermentation liquor, preliminary grinding and crushing by using grinding equipment, mixing of buffer solution, heating to 35 ℃, mixing of cellulase solution, enzymolysis and cooling for later use. This method based on polysaccharide is extracted to white fungus spore zymotic fluid, through smashing, extracting to the white fungus spore, can utilize the effective abundant of white fungus by the at utmost, guarantee extraction efficiency, the cellulase can realize hydrolysising the polysaccharide of macromolecule, guarantees to promote the extraction content of polysaccharide, guarantees the abundant raw and other materials that utilize, through extracting the white fungus spore solid production process in advance for inorganic salt is appeared in advance, adopts the water to be the solvent afterwards, dissolves inorganic salt, has reduced the preparation cost.
Description
Technical Field
The invention relates to the field of polysaccharide preparation and application, in particular to a method for extracting polysaccharide based on tremella spore fermentation liquor.
Background
The tremella polysaccharide is the main active component of tremella, comprises acidic polysaccharide, neutral heteropolysaccharide, acidic oligosaccharide, mural polysaccharide and exopolysaccharide, has effects of tonifying heart, delaying aging, resisting thrombi, resisting tumor, improving immunity and removing chloasma, has wide application in the fields of food, health products and cosmetics, and the tremella polysaccharide with high molecular weight has good moisture-keeping capacity and excellent film-forming property, and has the functions of oxidation resistance, allergy resistance and inflammation diminishing, but has the disadvantages of low solubility, difficult absorption by the skin and sticky feeling brought by application in the formula, the tremella polysaccharide with low molecular weight has certain defects in the application of cosmetics, and the tremella polysaccharide with low molecular weight has high solubility, is easy to be absorbed by skin, has good moisturizing capability and provides deep moisturizing effect for the skin, so the tremella polysaccharide with low molecular weight is more suitable for being applied to the field of cosmetics.
Spores (spore) are germ cells that, after leaving the parent, develop directly or indirectly into new individuals, which are the products of mitosis or meiosis; most of the spores are haploid and few of the spores are diploid, the spores are generally unicellular, and also can be multicellular propagules, and due to different characters, differences of the occurrence process and the structure, the diversity of the spores is formed.
At present, methods for extracting tremella polysaccharide from tremella sporophore or spore mainly comprise a hot water leaching method, a solvent extraction method, an enzyme dissolving extraction method and the like, the intracellular polysaccharide is dissolved out by destroying tremella cells, however, tremella spores not only contain polysaccharide, but also contain protein and inorganic salt, in the actual preparation process, a large amount of time and reagents are needed for removing the protein, a large amount of reagent residues are still generated, and secondly, in the subsequent preparation process, the inorganic salt is difficult to remove in advance, so that the problems of excessive impurity content of the polysaccharide and high actual preparation cost are caused.
Disclosure of Invention
The invention aims to provide a method for extracting polysaccharide based on tremella spore fermentation liquor. According to the method for extracting the polysaccharide based on the tremella spore fermentation liquid, disclosed by the invention, for the preparation efficiency of the tremella polysaccharide, the extracts are dissolved and precipitated by repeatedly adopting polyethylene glycol and ethanol, and the polyethylene glycol and the ethanol used in the production process are recycled, so that the waste of raw materials is avoided.
In order to achieve the above effects, the present invention provides the following technical solutions: a method for extracting polysaccharide based on tremella spore fermentation liquor comprises the following steps:
step one, raw material preparation, filtration of tremella spore fermentation liquor, preliminary grinding and crushing by using grinding equipment, mixing of buffer solution, heating to 35 ℃, mixing of cellulase solution, enzymolysis and cooling for later use.
And step two, performing coarse separation, taking out the enzymatic hydrolysate, heating, centrifuging, and performing freeze-drying treatment to obtain a solid product.
And step three, purification, mixing and soaking the solid product with absolute ethyl alcohol and polyethylene glycol, centrifuging, taking supernatant, and evaporating to obtain concentrated solution.
And step four, preparing polysaccharide, adding absolute ethyl alcohol again, precipitating with alcohol for 24 hours, and preparing freeze-dried powder through a hollow evaporator.
Further, the method comprises the following steps: according to the operation steps in the first step, the fermentation liquor is filtered, and the filtering diameter is the diameter of tremella spores.
Further, the method comprises the following steps: according to the operation steps in the first step, the buffer solution is prepared by dilute hydrochloric acid solution, the concentration of the dilute hydrochloric acid solution is 0.4-0.8mol/L, and the pH value of the buffer solution is 4-5.
Further, the method comprises the following steps: according to the operation steps in the second step, the heating temperature of the enzymolysis liquid is 40-60 ℃.
Further, the method comprises the following steps: according to the operating step in step two, the solid product is a mixture of protein and polysaccharide.
Further, the method comprises the following steps: according to the operation steps in the third step, the mixing and soaking time of the absolute ethyl alcohol and the polyethylene glycol needs to be more than 24 hours, and the concentrated solution is one tenth of the original volume.
Further, the method comprises the following steps: according to the operation steps in the third step, the centrifugation needs to be kept for more than 10 minutes, and the centrifugation time is increased by 5 minutes for every 1L of supernatant.
Further, the method comprises the following steps: according to the operation steps in the fourth step, the mixing ratio of the added absolute ethyl alcohol to the concentrated solution is 10: 1.
further, the method comprises the following steps: according to the operation steps in the fourth step, the alcohol precipitation process needs to be cooled for 48 hours, and then supernatant is taken to prepare freeze-dried powder.
Further, the method comprises the following steps: according to the operation steps in the fourth step, the freeze-dried powder is prepared by adopting the vacuum degree of 0.1-0.2MPa and the temperature of 28-38 ℃.
The invention provides a method for extracting polysaccharide based on tremella spore fermentation liquor, which has the following beneficial effects:
(1) according to the method for extracting the polysaccharide based on the tremella spore fermentation liquor, the tremella spores are crushed and extracted, so that the tremella is effectively and fully utilized to the greatest extent, and the extraction efficiency is ensured.
(2) The method for extracting the polysaccharide based on the tremella spore fermentation liquor can hydrolyze macromolecular polysaccharide by using the cellulase, so that the extraction content of the polysaccharide is improved, and the raw materials are fully utilized.
(3) According to the method for extracting the polysaccharide based on the tremella spore fermentation liquor, the tremella spore is extracted, the inorganic salt is precipitated in advance through the solid production treatment in advance, and then the water is used as a solvent to dissolve the inorganic salt, so that the preparation cost is reduced.
Drawings
FIG. 1 is a schematic view of the preparation method of the present invention.
Detailed Description
The invention provides a technical scheme that: referring to fig. 1, a method for extracting polysaccharide based on tremella spore fermentation broth includes the following steps:
step one, raw material preparation, filtration of tremella spore fermentation liquor, preliminary grinding and crushing by using grinding equipment, mixing of buffer solution, heating to 35 ℃, mixing of cellulase solution, enzymolysis and cooling for later use.
And step two, performing coarse separation, taking out the enzymatic hydrolysate, heating, centrifuging, and performing freeze-drying treatment to obtain a solid product.
And step three, purification, mixing and soaking the solid product with absolute ethyl alcohol and polyethylene glycol, centrifuging, taking supernatant, and evaporating to obtain concentrated solution.
And step four, preparing polysaccharide, adding absolute ethyl alcohol again, precipitating with alcohol for 24 hours, and preparing freeze-dried powder through a hollow evaporator.
Specifically, the method comprises the following steps: and (4) filtering the fermentation liquor according to the operation steps in the step one, wherein the filtering diameter is the diameter of tremella spores.
Specifically, the method comprises the following steps: according to the operation steps in the first step, the buffer solution is prepared by dilute hydrochloric acid solution, the concentration of the dilute hydrochloric acid solution is 0.4-0.8mol/L, and the pH value of the buffer solution is 4-5.
Specifically, the method comprises the following steps: according to the operation steps in the second step, the heating temperature of the enzymolysis liquid is 40-60 ℃.
Specifically, the method comprises the following steps: according to the operation in step two, the solid product is a mixture of protein and polysaccharide.
Specifically, the method comprises the following steps: according to the operation steps in the third step, the mixing and soaking time of the absolute ethyl alcohol and the polyethylene glycol needs to be more than 24 hours, and the concentrated solution is one tenth of the original volume.
Specifically, the method comprises the following steps: according to the operation steps in the third step, the centrifugation needs to be kept for more than 10 minutes, and the centrifugation time is increased by 5 minutes for every 1L of supernatant.
Specifically, the method comprises the following steps: according to the operation steps in the fourth step, the anhydrous ethanol and the concentrated solution are added again, wherein the mixing ratio of the anhydrous ethanol to the concentrated solution is 10: 1.
specifically, the method comprises the following steps: according to the operation steps in the fourth step and the alcohol precipitation process, after cooling for 48 hours, taking the supernatant to prepare the freeze-dried powder.
Specifically, the method comprises the following steps: according to the operation steps in the fourth step, the freeze-dried powder is prepared by adopting the vacuum degree of 0.1-0.2MPa and the temperature of 28-38 ℃.
The method of the examples was performed for detection analysis and compared to the prior art to yield the following data:
| efficiency of preparation | Cleanliness | Cost of preparation | |
| Examples | Is higher than | It is quite high | Is lower than |
| Prior Art | Is lower than | Is lower than | Is higher than |
According to the table data, when the embodiment is implemented, the method for extracting the polysaccharide based on the tremella spore fermentation liquid has the advantages that for the preparation efficiency of the tremella polysaccharide, polyethylene glycol and ethanol are repeatedly adopted for dissolving and precipitating the extract, and the polyethylene glycol and the ethanol used in the production process are recycled, so that the waste of raw materials is avoided.
The invention provides a method for extracting polysaccharide based on tremella spore fermentation liquor, which comprises the following steps: step one, preparing raw materials, filtering tremella spore fermentation liquor, then primarily grinding and crushing by using grinding equipment, mixing buffer solution, heating to 35 ℃, mixing cellulase solution, performing enzymolysis, cooling for later use, filtering the fermentation liquor, wherein the filtering diameter is the diameter of tremella spores, the buffer solution is prepared by dilute hydrochloric acid solution, the concentration of the dilute hydrochloric acid solution is 0.4-0.8mol/L, the pH value of the buffer solution is 4-5, filtering the fermentation liquor can realize the filtering of solid agar, peptone and other culture dish substances to avoid generating redundant impurities, step two, performing coarse separation, taking out the enzymolysis liquid, heating, centrifuging, performing freeze-drying treatment to obtain a solid product, the heating temperature of the enzymolysis liquid is 40-60 ℃, the solid product is a mixture of protein and polysaccharide, and solidifying the solid product, the method can avoid containing water, in the actual production process, the solid product can also contain inorganic salt, redissolution can be carried out through distilled water, the inorganic salt is removed, step three, purification is carried out, the solid product is mixed and soaked with absolute ethyl alcohol and polyethylene glycol, supernatant is taken after centrifugation, evaporation is carried out to be concentrated solution, the mixing and soaking time of the absolute ethyl alcohol and the polyethylene glycol needs to reach more than 24 hours, the concentrated solution is one tenth of the original volume, the centrifugation needs to be kept for more than 10 minutes, every 1L of supernatant is added, the centrifugation time is increased for 5 minutes, step four, polysaccharide is prepared, the absolute ethyl alcohol is added again, alcohol precipitation is carried out for 24 hours, freeze-dried powder preparation is carried out through a hollow evaporator, the mixing ratio of the absolute ethyl alcohol and the concentrated solution is 10: 1, carrying out an alcohol precipitation process, cooling for 48 hours, taking supernatant, and carrying out freeze-dried powder preparation, wherein the freeze-dried powder preparation adopts a vacuum degree of 0.1-0.2MPa and a temperature of 28-38 ℃.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A method for extracting polysaccharide based on tremella spore fermentation liquor is characterized by comprising the following steps:
s1, preparing raw materials, filtering tremella spore fermentation liquor, then primarily grinding and crushing by using grinding equipment, mixing buffer solution, heating to 35 ℃, mixing cellulase solution, performing enzymolysis, and cooling for later use;
s2, carrying out rough separation, taking out the enzymatic hydrolysate, heating, centrifuging, and then carrying out freeze-drying treatment to obtain a solid product;
s3, purifying, mixing and soaking the solid product with absolute ethyl alcohol and polyethylene glycol, centrifuging, taking supernate, and evaporating to obtain concentrated solution;
s4, preparing polysaccharide, adding absolute ethyl alcohol again, precipitating with ethanol for 24 hours, and preparing freeze-dried powder through a hollow evaporator.
2. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: and according to the operation step in the S1, filtering the fermentation liquor, wherein the filtering diameter is the diameter of tremella spores.
3. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the procedure in S1, the buffer solution is prepared as a dilute hydrochloric acid solution having a concentration of 0.4 to 0.8mol/L and a pH of 4 to 5.
4. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the operation step in S2, the heating temperature of the enzymolysis liquid is 40-60 ℃.
5. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the procedure in S2, the solid product is a mixture of protein and polysaccharide.
6. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the operation step in S3, the mixing and soaking time of the absolute ethyl alcohol and the polyethylene glycol needs to be more than 24 hours, and the concentrated solution is one tenth of the original volume.
7. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the procedure in S3, the centrifugation needs to be maintained for 10 minutes or more, and the centrifugation time is increased by 5 minutes for every 1L of the supernatant.
8. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the operation procedure in S4, the mixing ratio of the added absolute ethyl alcohol to the concentrated solution is 10: 1.
9. the method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the operation steps in S4, in the alcohol precipitation process, after cooling for 48 hours, supernatant is taken for freeze-dried powder preparation.
10. The method for extracting polysaccharide based on tremella spore fermentation broth as claimed in claim 1, comprising the following steps: according to the operation steps in S4, the freeze-dried powder is prepared under the conditions that the vacuum degree is 0.1-0.2MPa and the temperature is 28-38 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110721695.2A CN113583145A (en) | 2021-06-28 | 2021-06-28 | Method for extracting polysaccharide based on tremella spore fermentation liquor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110721695.2A CN113583145A (en) | 2021-06-28 | 2021-06-28 | Method for extracting polysaccharide based on tremella spore fermentation liquor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113583145A true CN113583145A (en) | 2021-11-02 |
Family
ID=78244859
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110721695.2A Pending CN113583145A (en) | 2021-06-28 | 2021-06-28 | Method for extracting polysaccharide based on tremella spore fermentation liquor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113583145A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115819638A (en) * | 2022-12-28 | 2023-03-21 | 广东药科大学 | Preparation method of tremella polysaccharides with different molecular weights |
-
2021
- 2021-06-28 CN CN202110721695.2A patent/CN113583145A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115819638A (en) * | 2022-12-28 | 2023-03-21 | 广东药科大学 | Preparation method of tremella polysaccharides with different molecular weights |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20220080333A1 (en) | Ultrasonic composite acidic water extraction method for cordyceps polysaccharide and cordycepin in cordyceps militaris | |
| CN101323649A (en) | Method for extracting mushroom polysaccharide by using ultra-high pressure | |
| CN109608559A (en) | A method of extracting active polysaccharide from seaweed | |
| CN107868805B (en) | Longan polysaccharide degraded by lactobacillus fermentation and preparation method thereof | |
| CN111019011B (en) | Method for extracting rice bran polysaccharide | |
| CN105254778A (en) | Extracting method of sisal hemp pectin | |
| CN104382953A (en) | Extraction method of melanin in black fungi | |
| CN103271951A (en) | Method for preparing cordyceps preparations with high adenosine contents | |
| CN113583145A (en) | Method for extracting polysaccharide based on tremella spore fermentation liquor | |
| CN101225421A (en) | Novel process for extracting golden fungus polysaccharides by step enzyme method | |
| CN107267571A (en) | A kind of extracting method of rice bran polysaccharide | |
| JP2017521451A (en) | Method for extracting soluble proteins from microalgal biomass | |
| CN104292355A (en) | Method for extraction of platycodon grandiflorum polysaccharide | |
| CN109758531A (en) | High-fidelity, Herba Dendrobii extract of high clarity and preparation method thereof | |
| CN112175098A (en) | Method for efficiently preparing polysaccharide from sparassis crispa stipe | |
| KR101687985B1 (en) | Subcritical extraction method of -glucan derived from mushroom | |
| CN110484577B (en) | Method for extracting and preparing mannose from dragon fruit stems | |
| JPH06113871A (en) | Method for separating and recovering flavonoid compound and limonoid compound contained in citrus fruits | |
| CN112940146A (en) | Preparation method of tremella polysaccharide | |
| CN113694561A (en) | Extraction and separation method of elderberry polyphenol | |
| CN113563488A (en) | Preparation method of pharmaceutical-grade micromolecular marine organism polysaccharide | |
| CN106947796A (en) | A kind of D trehaloses purifying technique | |
| CN107141365B (en) | Method for efficiently purifying phellinus igniarius polysaccharides by repeatedly increasing and decreasing pressure | |
| CN110863024A (en) | Method for preparing micromolecular hyaluronic acid by utilizing squid eyes | |
| CN117247849B (en) | Microalgae extraction method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211102 |