CN110437343A - A kind of extracting mode of lentinan - Google Patents
A kind of extracting mode of lentinan Download PDFInfo
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- CN110437343A CN110437343A CN201910640763.5A CN201910640763A CN110437343A CN 110437343 A CN110437343 A CN 110437343A CN 201910640763 A CN201910640763 A CN 201910640763A CN 110437343 A CN110437343 A CN 110437343A
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- Prior art keywords
- lentinan
- glycerine
- choline chloride
- phase
- k2hpo4
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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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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
The invention discloses a kind of extracting mode of lentinan, S1, material prepares；S2, the preparation of lentinan Aqueous extracts；S3, the synthesis of choline chloride/glycerine eutectic solvent；The preparation of S4, K2HPO4 solution；S5, the building of choline chloride/glycerine-salt double-aqueous phase system；S6, the removing of protein: to the K2HPO4 solution being added in graduated centrifuge tube in choline chloride/glycerine and S4 in S3, and the lentinan Aqueous extracts in S2, centrifugal treating is carried out using centrifuge, centrifuge tube is taken out after operation and is kept completely separate to two-phase；Protein in lentinan is sloughed using a kind of completely new method, choline chloride/glycerine eutectic solvent-salt double-aqueous phase system is constructed in conjunction with suitable inorganic salts using the wide eutectic solvent of recent years development prospect to extract protein from lentinan, and the lentinan rate of recovery of the invention is 98.0%, is up to 90.8% to the removal efficiency of lentinan protein matter.
Description
Technical field
The present invention relates to lentinan extractive technique field, specially a kind of extracting mode of lentinan.
Background technique
Mushroom, scientific name Lentinula edodes (Berk.) Sing.Mushroom is in the side such as color, taste, mouthfeel, nutrition
Face is more unique compared with compared with his mushroom, is edible mushroom preferred by everyone, there is great demand on world market.
The fresh mushroom and dried thin mushroom of common sale have different mouthfeels in the market, and with the development of dehydration technique, mushroom is more
It is to be processed to the clear-cut piece of mushroom, becomes a kind of snacks product of health-nutrition.Mushroom not only delicious flavour is also rich in albumen
The nutriments such as matter, polysaccharide, cellulose and multivitamin, amino acid.Mushroom can delay senescence, and blood pressure is inhibited to rise, and resist
Cancer is antitumor, and can enhance the immunity function of body.
Lentinan is polarity macromolecular compound, however in the lentinan obtained with hot water or alkaline extraction usually
It is mingled with protein.The method of common removing protein has Sevag method, trifluorotrichloroethane method, trichloroacetic acid method etc..Sevag method is multiple
Miscellaneous, time-consuming and sample loss is larger, and trifluorotrichloroethane method and trichloroacetic acid method are both needed to carry out at a lower temperature, is not easy to
The progress of operator.
Summary of the invention
The purpose of the present invention is to provide a kind of extracting modes of lentinan, mentioned above in the background art to solve
Problem.
To achieve the above object, the invention provides the following technical scheme: a kind of extracting mode of lentinan, including it is following
Step:
S1, material prepare: enough fresh and dried mushroom and choline chloride, glycerine and K2HPO4 are bought on the market, purchase
Fresh and dried mushroom cleans stripping and slicing.
S2, the preparation of lentinan Aqueous extracts: taking the fresh and dried mushroom of stripping and slicing in S1, then suitably crushed, and is added suitable
The tap water of amount, then water-bath refluxing extraction is taken, after the completion of extraction, lentinan Aqueous extracts are filtered to obtain, refrigerator cold-storage is placed in
Room is spare.
The synthesis of choline chloride/glycerine eutectic solvent: S3 takes choline chloride appropriate and appropriate glycerine, the two object
The amount ratio of matter is 1:2, is placed in dry vessel and is mixed, and water-bath heated at constant temperature and cooperative mechanical stirring obtain colorless and transparent
Liquid, be stored under room temperature environment store it is spare.
The preparation of S4, K2HPO4 solution: taking a certain amount of K2HPO4 to be dissolved in distilled water, is configured to K2HPO4 solution.
Choline chloride/glycerine-salt double-aqueous phase system building: S5 is poured into test tube and is synthesized in a certain amount of S3
Choline chloride/glycerine eutectic solvent adds the K2HPO4 solution configured in appropriate S4, is then uniformly rocked, and protects
It is constant to hold ambient temperature, sees whether to become clearly two-phase, the then building success of two-phase cleaning if two-phase is unintelligible, is rushed
Configuration choline chloride/glycerine and K2HPO4 solution are washed, until two-phase is clear, retains two-phase clearly choline chloride/glycerine
Eutectic solvent and K2HPO4 solution, it is spare.
S6, the removing of protein: in choline chloride/glycerine and S4 in graduated centrifuge tube in addition S3
Lentinan Aqueous extracts in K2HPO4 solution and S2 carry out centrifugal treating using centrifuge, will centrifugation after operation
Pipe taking-up is kept completely separate to two-phase.
Preferably, the mushroom that wherein treated in S1 is stored in crisper, institute spare as storing under low temperature environment
Its temperature of the low temperature environment stated controls in the range of 1 DEG C to 4 DEG C.
Preferably, wherein the control of water-bath reflux extracting time is controlled in 2h, the temperature of refrigerating chamber of refrigerator at -2 DEG C in S2
To -4 DEG C.
Preferably, wherein the temperature of water-bath is set as 80 DEG C in S3, and churned mechanically time control therein exists
30min。
Preferably, the K2HPO4 solution being wherein made in S4 be stored in room temperature environment store it is spare.
Preferably, the revolving speed that wherein centrifuge sets centrifuge in S6 be set as the 2000r/min Centrifugical extraction time as
30min。
Preferably, wherein two-phase liquid extraction obtained is a certain amount of in S6, is stored in two different vessel respectively, and
Polysaccharide is measured using phenol-sulphuric acid colorimetry afterwards, and protein is measured using Coomassie Brilliant Blue, then in two upper phases
It is respectively dropped into the concentrated sulfuric acid and Coomassie brilliant G-250 in liquid, then will instill phenol in the vessel for instilling the concentrated sulfuric acid again, observes
It develops the color if being instilled in the solution in upper phase liquid in the vessel of phenol, upper phase liquid is the polysaccharide for needing to extract, if otherwise instilling
Cyan is shown in the vessel of Coomassie brilliant G-250, then upper phase liquid is protein, and lower phase liquid is the polysaccharide for needing to extract.
Preferably, wherein for the substances such as polysaccharide under the action of the concentrated sulfuric acid, glycosidic bond fracture, polysaccharide hydrolysis is at monosaccharide, monosaccharide
Dehydration generates furfural and its derivative, these substances act on phenol generate coloring matter again, and Coomassie brilliant G-250 itself is
Red has maximum light absorption value under 465nm wavelength, and when in conjunction with protein, its color is changed into cyan by red.
Compared with prior art, the beneficial effects of the present invention are:
The present invention sloughs the protein in lentinan using a kind of completely new method.Before recent years research hot topic, development
The wide eutectic solvent of scape constructs choline chloride/glycerine eutectic solvent-salt aqueous two-phase body in conjunction with suitable inorganic salts
System extracts protein from lentinan, and the lentinan rate of recovery of the invention is 98.0%, to lentinan egg
The removal efficiency of white matter is up to 90.8%, and choline chloride/glycerine eutectic solvent is as a kind of environmentally friendly ionic liquid
Body analog has many advantages, such as that property is stable, selectivity is strong, cheap.
Figure of description
Fig. 1 is choline chloride/glycerine eutectic solvent-salt double-aqueous phase system extracting protein matter schematic diagram.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
A kind of extracting mode of lentinan, comprising the following steps:
S1, material prepare: enough fresh and dried mushroom and choline chloride, glycerine and K2HPO4 are bought on the market, purchase
Fresh and dried mushroom cleans stripping and slicing, and the mushroom that treated is stored in crisper spare as storing under low temperature environment, and described is low
Its temperature of warm environment controls in the range of 1 DEG C to 4 DEG C.
S2, the preparation of lentinan Aqueous extracts: taking the fresh and dried mushroom of stripping and slicing in S1, then suitably crushed, and is added suitable
The tap water of amount, then water-bath refluxing extraction is taken, after the completion of extraction, lentinan Aqueous extracts are filtered to obtain, refrigerator cold-storage is placed in
Room is spare, and the control of water-bath reflux extracting time is controlled in 2h, the temperature of refrigerating chamber of refrigerator at -2 DEG C to -4 DEG C.
The synthesis of choline chloride/glycerine eutectic solvent: S3 takes choline chloride appropriate and appropriate glycerine, the two object
The amount ratio of matter is 1:2, is placed in dry vessel and is mixed, and water-bath heated at constant temperature and cooperative mechanical stirring obtain colorless and transparent
Liquid is stored under room temperature environment and store spare, and the temperature of water-bath is set as 80 DEG C, therein churned mechanically
Time controls in 30min.
The preparation of S4, K2HPO4 solution: taking a certain amount of K2HPO4 to be dissolved in distilled water, is configured to K2HPO4 solution, middle system
Obtained K2HPO4 solution be stored in room temperature environment store it is spare.
Choline chloride/glycerine-salt double-aqueous phase system building: S5 is poured into test tube and is synthesized in a certain amount of S3
Choline chloride/glycerine eutectic solvent adds the K2HPO4 solution configured in appropriate S4, is then uniformly rocked, and protects
It is constant to hold ambient temperature, sees whether to become clearly two-phase, the then building success of two-phase cleaning if two-phase is unintelligible, is rushed
Configuration choline chloride/glycerine and K2HPO4 solution are washed, until two-phase is clear, retains two-phase clearly choline chloride/glycerine
Eutectic solvent and K2HPO4 solution, it is spare.
S6, the removing of protein: in choline chloride/glycerine and S4 in graduated centrifuge tube in addition S3
Lentinan Aqueous extracts in K2HPO4 solution and S2 carry out centrifugal treating using centrifuge, will centrifugation after operation
Pipe taking-up is kept completely separate to two-phase, and the revolving speed that centrifuge sets centrifuge is set as the 2000r/min Centrifugical extraction time as 30min.
Choline chloride/glycerine eutectic solvent-salt double-aqueous phase system extracting protein matter schematic diagram is as shown in Figure 1.
Wherein two-phase liquid extraction obtained is a certain amount of in S6, is stored in two different vessel respectively, then uses phenol-
Sulfuric acid development process measures protein to measure polysaccharide, and using Coomassie Brilliant Blue, then drips respectively in two upper phase liquid
Enter the concentrated sulfuric acid and Coomassie brilliant G-250, then will instill phenol in the vessel for instilling the concentrated sulfuric acid again, if the upper phase liquid of observation
In solution in instill in the vessel of phenol and develop the color, upper phase liquid be to need the polysaccharide that extracts, if instillation Coomassie brilliant blue on the contrary
Cyan is shown in the vessel of G-250, then upper phase liquid is protein, and lower phase liquid is the polysaccharide for needing to extract, and the substances such as polysaccharide are in dense sulphur
Under the action of acid, glycosidic bond fracture, for polysaccharide hydrolysis at monosaccharide, monosaccharide dehydration generates furfural and its derivative, these substances again with
Phenol effect generate coloring matter, Coomassie brilliant G-250 itself be red, have maximum light absorption value under 465nm wavelength, when with
When protein combines, its color is changed into cyan by red.
It should be noted that, in this document, relational terms such as first and second and the like are used merely to a reality
Body or operation are distinguished with another entity or operation, are deposited without necessarily requiring or implying between these entities or operation
In any actual relationship or order or sequence.Moreover, the terms "include", "comprise" or its any other variant are intended to
Non-exclusive inclusion, so that the process, method, article or equipment including a series of elements is not only wanted including those
Element, but also including other elements that are not explicitly listed, or further include for this process, method, article or equipment
Intrinsic element.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (8)
1. a kind of extracting mode of lentinan, which comprises the following steps:
S1, material prepare: enough fresh and dried mushroom and choline chloride, glycerine and K2HPO4 are bought on the market, purchase
Fresh and dried mushroom cleans stripping and slicing；
S2, the preparation of lentinan Aqueous extracts: taking the fresh and dried mushroom of stripping and slicing in S1, then suitably crushed, and is added suitable
Tap water, then water-bath refluxing extraction is taken, after the completion of extraction, lentinan Aqueous extracts are filtered to obtain, it is standby to be placed in cold compartment of refrigerator
With；
S3, the synthesis of choline chloride/glycerine eutectic solvent: taking choline chloride appropriate and appropriate glycerine, the two substance
Amount is placed in dry vessel and is mixed, water-bath heated at constant temperature and cooperative mechanical stirring obtain colourless transparent liquid than being 1:2
, be stored under room temperature environment store it is spare；
The preparation of S4, K2HPO4 solution: taking a certain amount of K2HPO4 to be dissolved in distilled water, is configured to K2HPO4 solution；
The building of choline chloride/glycerine-salt double-aqueous phase system: S5 pours into the chlorination synthesized in a certain amount of S3 into test tube
Choline/glycerine eutectic solvent adds the K2HPO4 solution configured in appropriate S4, is then uniformly rocked, and keeps outer
Boundary is temperature-resistant, sees whether to become clearly two-phase, the then building success of two-phase cleaning, if two-phase is unintelligible, flushing is matched
Choline chloride/glycerine and K2HPO4 solution are set, until two-phase is clear, retaining two-phase, clearly choline chloride/glycerine is low total
Molten solvent and K2HPO4 solution for standby；
S6, the removing of protein: in choline chloride/glycerine and S4 in graduated centrifuge tube in addition S3
Lentinan Aqueous extracts in K2HPO4 solution and S2 carry out centrifugal treating using centrifuge, will centrifugation after operation
Pipe taking-up is kept completely separate to two-phase.
2. a kind of extracting mode of lentinan according to claim 1, it is characterised in that: what wherein treated in S1
Mushroom be stored in crisper as stored under low temperature environment its temperature of spare, described low temperature environment control at 1 DEG C to 4 DEG C
In range.
3. a kind of extracting mode of lentinan according to claim 1, it is characterised in that: wherein water-bath reflux mentions in S2
Take time control in 2h, the temperature of refrigerating chamber of refrigerator is controlled at -2 DEG C to -4 DEG C.
4. a kind of extracting mode of lentinan according to claim 1, it is characterised in that: wherein in S3 water-bath temperature
Degree is set as 80 DEG C, and churned mechanically time control therein is in 30min.
5. a kind of extracting mode of lentinan according to claim 4, it is characterised in that: be wherein made in S4
K2HPO4 solution be stored in room temperature environment store it is spare.
6. a kind of extracting mode of lentinan according to claim 1, it is characterised in that: wherein centrifuge is set in S6
The revolving speed of centrifuge is set as the 2000r/min Centrifugical extraction time as 30min.
7. a kind of extracting mode of lentinan according to claim 1, it is characterised in that: wherein two-phase obtained in S6
Liquid extraction is a certain amount of, is stored in two different vessel respectively, then measures polysaccharide using phenol-sulphuric acid colorimetry,
And protein is measured using Coomassie Brilliant Blue, then the concentrated sulfuric acid and Coomassie brilliant blue are respectively dropped into two upper phase liquid
G-250 then will instill phenol in the vessel for instilling the concentrated sulfuric acid again, if instilling the device of phenol in the solution in the upper phase liquid of observation
It develops the color in ware, upper phase liquid is the polysaccharide for needing to extract, if showing cyan in the vessel on the contrary for instilling Coomassie brilliant G-250, then
Upper phase liquid is protein, and lower phase liquid is the polysaccharide for needing to extract.
8. a kind of extracting mode of lentinan according to claim 7, it is characterised in that: wherein the substances such as polysaccharide are dense
Under the action of sulfuric acid, glycosidic bond fracture, polysaccharide hydrolysis generates furfural and its derivative at monosaccharide, monosaccharide dehydration, these substances are again
It being acted on phenol and generates coloring matter, Coomassie brilliant G-250 itself is red, there is maximum light absorption value under 465nm wavelength, when
When in conjunction with protein, its color is changed into cyan by red.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113307890A (en) * | 2021-06-04 | 2021-08-27 | 山西农业大学 | Morchella polysaccharide and deep eutectic solvent extraction method thereof |
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