CN110272932A - A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide - Google Patents
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide Download PDFInfo
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Abstract
The invention belongs to field of biotechnology, and in particular to a kind of preparation method of lucid ganoderma spore powder polysaccharide peptide.The preparation method of lucid ganoderma spore powder polysaccharide peptide of the invention includes carrying out broken wall treatment to lucidum spore powder, being extracted using supercritical carbon dioxide, digested to degreasing ganoderma spove powder, extracting filtering and concentration, drying, the process that pulverizes and sieves to enzymolysis liquid.Process of the invention is easier, green safe, is convenient for industrialized production；Time shortening is handled, processing overlong time ganoderma lucidum functional component deactivation is prevented；Glycopeptide structure will not be destroyed, be easy filtering by digesting the enzyme used；Organic solvent is not used, can be avoided organic solvent residual problem.In addition, by means of the present invention, glycopeptide yield can reach 9.50%, and the lucid ganoderma spore powder polysaccharide peptide content of sample is 36.11%.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of preparation method of lucid ganoderma spore powder polysaccharide peptide.
Background technique
Ganoderma lucidum is Basidiomycetes Polyporaceae Ganoderma fungi, and it is with a long history to eat rare Chinese medicine as traditional medicine.
Conidia powder is the spore constantly ejected from tube after ganoderma lucidum fruitbody is reached maturity, biologically referred to as basidiospore, is concentrated
In powdered after getting up, referred to as lucidum spore powder.Conidia powder summarizes the essential part of ganoderma lucidum, has more richer than ganoderma lucidum three
The ingredients such as terpene, polysaccharide, adenosine, protein, enzyme, selenium element.Largely studies have shown that lucidum spore powder has enhancing
Immunity of organisms inhibits the multiple pharmacological effects such as tumour, hypoglycemic, reducing blood lipid, protect liver, antiviral, anti-radiation.Reishi sporule tool
There is double-walled construction, is surrounded by hard chitin fiber, quality is hard, acid and alkali-resistance, hinders human body to its effective component
It is absorbed and utilized.In order to make full use of the effective component in lucidum spore powder, it is necessary to broken wall is carried out to conidia powder, in order to have to it
Imitate the utilization of ingredient.
A variety of pharmacological activity of ganoderma lucidum all have significant relationship with ganoderma polysaccharide peptide mostly.Ganoderma polysaccharide peptide is a kind of sugared egg
It is white, it is the combination of polysaccharide and protein, is made of the repetitive unit of disaccharide, typically contains very a high proportion of polysaccharide.Current
Studies have shown that ganoderma polysaccharide peptide has extensive pharmacological activity, the immunity of human body can be improved, improve the hypoxia-bearing capability of body,
Interior free yl is eliminated, tumour cell is inhibited to generate, it is anti-radiation, improve liver, marrow, blood synthetic DNA, RNA and protein
Ability, the health-care efficacy for playing anti-aging, extending the service life.In addition, ganoderma polysaccharide peptide also has non-specificity in human activin
Antibody, the immune idiosyncrasy of enhancing, inhibition transplantation tumor physiological activity and other effects.Ganoderma polysaccharide peptide to cardiovascular disease, asthma,
Allergy, neurasthenia, stomach energy etc. have significant effect, also have lowering blood pressure and blood fat, solution blood stasis, improve blood circulation, whitening
The effects of skin.
Currently, ganoderma polysaccharide peptide mainly using ganoderma lucidum mycelium, ganoderma lucidum fruitbody as raw material, is mentioned by water, alkali carries, alcohol extracting etc.
Extraction process obtains lucidum extracting liquid, then by the films filtration treatment such as refined filtration, ultrafiltration, reverse osmosis, then with zwitterion resin
One of column chromatography methods such as column, gel column, middle compression leg, reversed-phase column or multiple combinations, or do not fill column only in column resin,
The filler combinations such as silica gel, gel carry out purification process to lucidum extracting liquid.These methods by repeatedly, the film of different principle
Filtering, column, which chromatograph, to be achieved the purpose that isolate and purify ganoderma polysaccharide peptide, and obtain the higher product of purity, property is generally more stable.So
And the loss of these processes is big, equipment disposably puts into that big, equipment clean and maintenance expense is costly and time consuming, the production time is long, production
It is at high cost, it needs in extraction process using a large amount of organic solvents, leading to product, there may be dissolvent residuals, give product edible safety
Aspect brings hidden danger.In addition, that there is also extraction efficiencies is low, extraction cost is high, product matter for existing ganoderma polysaccharide peptide extraction purification
Measure the problems such as lower, active constituent content is lower, stability is poor.For example, existing extracting method isolates and purifies work by multiple
Sequence finally obtains high-purity ganoderma polysaccharide peptide, but production process is many and diverse, need using it is numerous it is high it is profound isolate and purify equipment,
It puts into big, at high cost, it is difficult to realize industrialized production, product is also difficult to popularize application.
CN201610221431.X discloses a kind of ganoderma lucidum fruitbody and the comprehensive extraction and application production method of Reishi sporule.It should
Method needs to be depressured separation after carrying out supercritical extract to Reishi sporule, shows that parameter setting is unreasonable when supercritical extract, leads
Ganoderma lucidum spore oil in conidia powder is caused to fail sufficiently to extract；Ganoderma lucidum fruitbody is mixed with extract remainder and carries out water body, and ganoderma lucidum
Fructification degree of lignification is high, structure is complicated, and polysaccharide is generally incorporated in one by physically or chemically key with cellulose, lignin
It rises, is difficult to come out Polyose extraction using the method that water mentions, need to be extracted by multi-reflow；Ethanol precipitation is carried out, due to small point
Sub- polysaccharide is dissolved in ethyl alcohol, therefore amount of activated polysaccharide can be lost in alcohol precipitation, and alcohol hypostasis uses acetone washing, and it is residual that there are organic solvents
Stay problem.In fact, there are apparent defects for this method, such as production time length, high production cost, production process is many and diverse, extracts
There are residual solvents etc. for low efficiency, product.
CN200610097229.7 discloses a kind of process for refining of ganoderma spore polysaccharide, is extracted, is mentioned using warm leaching method water
It takes process to take a long time, the standing long period need to be needed to be filtered concentration again after combined extract by repeatedly extracting；Using alcohol
Heavy step, since micromolecular polysaccharide is dissolved in ethyl alcohol, amount of activated polysaccharide can be lost in alcohol precipitation；Subtractive process include decoloration,
Sevag method takes off albumen, dialysis, alcohol precipitation, washing, cumbersome, need using a large amount of organic solvents (such as sevag method take off albumen, add
Enter isometric chloroform-n-butanol (4:1), precipitating uses dehydrated alcohol and acetone washing 3 times after alcohol precipitation), there are organic solvents
Residue problem.Though the polysaccharide of ganoderma spore polysaccharide highly finished product contains in addition, being improved by polysaccharide yield after purification and polyoses content
Amount is calculated with DEXTROSE ANHYDROUS, not can prove that it for micromolecular polysaccharide peptide.
CN201610853239.2 discloses the preparation method of lucidum spore powder glycopeptide extract, successively includes dense alcohol extracting
It takes, dilute alcohol extraction, the ether defatting for merging dense alcohol extract and dilute alcohol extracting solution, removal of impurities decoloration, purifying resin step, and preferably
To after dense alcohol extracting, dilute alcohol are extracted, remaining lucidum spore powder dregs of a decoction further progress includes water extraction, alcohol precipitates and purification
The processing of step.This method uses a large amount of alcohol in extraction process, need to carry out repeatedly extracting reflux, just using ether defatting, chloroform-
Butanol (4:1) carries out purifying purification, and product is easy residual organic solvent, and process is complicated, and the production time is long, and production cost is higher,
It is difficult to industrialize in realization.
Summary of the invention
For problem of the prior art, the present invention provides a kind of extracting methods of lucid ganoderma spore powder polysaccharide peptide.The present invention
Using degreasing conidia powder as raw material, by it is feasible, suitable for industrialized method obtain the stable ganoderma polysaccharide peptide of property, will take off
Rouge conidia powder turns waste into wealth, and solves in the prior art that ganoderma polysaccharide peptide extraction efficiency is low, extraction cost is high and final products
The low problem of middle ganoderma polysaccharide peptide functional component inactivation, content, improves the effective rate of utilization of lucidum spore powder, to promotion ganoderma lucidum
The development of industry plays huge impetus.
The present invention is achieved by the following technical solutions:
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: low-temperature physics broken wall treatment is carried out to lucidum spore powder, sporoderm-broken rate is made to reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extraction apparatus and is extracted, then
It is separated twice, collects grease and degreasing ganoderma spove powder；
(3) digest: degreasing ganoderma spove powder is mixed with pure water, add aqueous slkali adjusting pH to 5-10, then plus
Enter enzyme to be digested；
(4) it extracts filtering: enzymolysis liquid being warming up to 90-100 DEG C under stirring condition and keeps the temperature 1-2h, obtains extracting solution, so
After separate, collect clear filtrate；
(5) be concentrated: filtrate is concentrated into the medicinal extract of solid content 35-55%；
(6) dry: vacuum drying medicinal extract；
(7) it pulverizes and sieves: smashing it through 60-80 mesh screen, obtain lucid ganoderma spore powder polysaccharide peptide powder.
In the preferred embodiment of the invention, the low-temperature physics broken wall is carried out using vibration type ultramicro disintegrator.Specifically
Lucidum spore powder is fitted into vibration type ultramicro disintegrator by ground, opening cooling water switch, after constant to 3-5 DEG C of water temperature to be cooled
Start broken wall, broken time 25-35min makes sporoderm-broken rate reach 95% or more.
In the preferred embodiment of the invention, the pumping frequency of the extraction equipment is 25-30Hz, the stream of supercritical carbon dioxide
Amount >=120L/h, extracting pressure 18-25MPa, extraction temperature are 40-50 DEG C, extraction time 4-5h.It is highly preferred that super face
The flow of boundary's carbon dioxide is 120-160L/h.
It is separated twice in the preferred embodiment of the invention, wherein level-one separating pressure is 8-10MPa, separation temperature
Degree is 40-50 DEG C；The second-order separation pressure is 5-6MPa, and separation temperature is 28-35 DEG C.
In the preferred embodiment of the invention, the concentration of the aqueous slkali is 2-3mol/L, and the aqueous slkali is hydroxide
One of sodium, potassium hydroxide, calcium hydroxide and ammonium hydroxide.
In the preferred embodiment of the invention, aqueous slkali is added to adjust pH to 7.0-8.5.
In the preferred embodiment of the invention, the enzyme is cellulase, compound protease, alkali protease and neutral egg
One of white enzyme is a variety of；The additional amount of the enzyme is the 0.1-2% of degreasing ganoderma spove powder weight.
In the preferred embodiment of the invention, the temperature of enzymatic hydrolysis is 40-55 DEG C, and the time of enzymatic hydrolysis is 2-12h.More preferably
Ground, the time of enzymatic hydrolysis are 4-10h.
It in the preferred embodiment of the invention, is ultrasonically treated during enzymatic hydrolysis, the frequency of ultrasonic treatment is 20-
40KHz, time 4-6h.
In the preferred embodiment of the invention, concentration is carried out using economic benefits and social benefits vacuum energy-saving concentrator, economic benefits and social benefits vacuum energy-saving
One effect vacuum degree of concentrator is -0.06Mpa, and temperature is 70-80 DEG C；Two effect vacuum degrees are -0.08MPa, temperature 50-60
℃。
In the preferred embodiment of the invention, vacuum drying temperature be 60-70 DEG C, pressure be -0.07 to -
0.098MPa。
Beneficial effects of the present invention:
The present invention uses degreasing ganoderma spove powder for raw material, by broken wall, degreasing, enzymatic hydrolysis, extraction, concentration, drying,
It crushes, sieving process acquisition lucid ganoderma spore powder polysaccharide peptide, entire method operating method is simple, and mild condition, production cost is low, mentions
Take it is high-efficient, can industrialized production effectively functional component in degreasing conidia powder can be extracted without using organic solvent
Come, be translated into the high lucid ganoderma spore powder polysaccharide peptide of added value, turn waste into wealth, greatly extends the sharp again of degreasing conidia powder
With.
Method compared with the prior art, the present invention is without the use of fine filterings sides such as reverse osmosis, nanofiltration, ultrafiltration, micro-filtrations
Method carries out purification process, saves the expense of equipment investment and working service；Process is easier, green safe, convenient for industrial metaplasia
It produces；Time shortening is handled, processing overlong time ganoderma lucidum functional component deactivation is prevented；Glycopeptide will not be destroyed by digesting the enzyme used
Structure is easy filtering；Organic solvent is not used, can be avoided organic solvent residual problem.In addition, the direct water of degreasing conidia powder
It mentions, glycopeptide yield is only 0.21%, and the lucid ganoderma spore powder polysaccharide peptide content of sample is only 1.95%；And side through the invention
Method, glycopeptide yield can reach 9.50%, and the lucid ganoderma spore powder polysaccharide peptide content of sample is 36.11%.
Specific embodiment
Below with reference to embodiment, the invention will be further described.
Embodiment 1
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 120L/h extracts 4h under conditions of pumping frequency 27.5Hz, extracting pressure 21MPa, temperature 45 C, so
It is separated, is collected under the conditions of level-one separating pressure 9MPa, temperature 45 C and the second-order separation pressure 5.5MPa, 31 DEG C of temperature afterwards
Grease and degreasing ganoderma spove powder；
(3) it digests: 50kg degreasing ganoderma spove powder and 500kg pure water is mixed, 2mol/ is added under stirring condition
L sodium hydroxide solution adjusts pH to 7.2, and the compound protease of cellulase and 0.25kg that 0.5kg is then added carries out enzyme
It solves, be kept stirring during enzymatic hydrolysis and maintain pH value of solution 7.2 ± 0.1, enzyme at a temperature of 48 DEG C by the way that 2mol/L sodium hydroxide is added
Solve 4h；
(4) it extracts separation: enzymolysis liquid being warming up to 95 DEG C under stirring condition and keeps the temperature 2h, extracting solution is obtained, then uses
Centrifuge 5000rpm revolving speed by extracting solution slag charge and liquid separate, collect clear filtrate；
(5) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 45%；
(6) dry: medicinal extract 12h is dried in vacuo under the conditions of 65 DEG C of temperature and pressure -0.084MPa；
(7) it pulverizes and sieves: smashing it through 80 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Embodiment 2
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 140L/h extracts 5h under conditions of pumping frequency 25Hz, extracting pressure 18MPa, temperature 50 C, then
It is separated under the conditions of level-one separating pressure 8MPa, temperature 50 C and the second-order separation pressure 5MPa, 35 DEG C of temperature, collects grease
With degreasing ganoderma spove powder；
(3) it digests: 50kg degreasing ganoderma spove powder and 300kg pure water is mixed, be added under stirring condition
2.5mol/L potassium hydroxide solution adjusts pH to 8.5, and the cellulase of 0.25kg and the basic protein of 0.325kg is then added
Enzyme is digested, and is kept stirring during enzymatic hydrolysis and is maintained pH value of solution 8.5 ± 0.1, ultrasound by the way that 2.5mol/L potassium hydroxide is added
Under the conditions of digest 4h at a temperature of 42 DEG C；
(4) it extracts separation: enzymolysis liquid being warming up to 95 DEG C under stirring condition and keeps the temperature 2h, extracting solution is obtained, then passes through
Flame filter press filtering, filtrate regather after being back to clarification；
(5) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 55%；
(6) dry: medicinal extract 10h is dried in vacuo under the conditions of temperature 70 C and pressure -0.07MPa；
(7) it pulverizes and sieves: smashing it through 60 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Embodiment 3
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 160L/h extracts 4h under conditions of pumping frequency 30Hz, extracting pressure 25MPa, 40 DEG C of temperature, then
It is separated under the conditions of level-one separating pressure 10MPa, 40 DEG C of temperature and the second-order separation pressure 6MPa, 28 DEG C of temperature, collects oil
Rouge and degreasing ganoderma spove powder；
(3) it digests: 50kg degreasing ganoderma spove powder and 250kg pure water is mixed, be added under stirring condition
3.0mol/L sodium hydroxide solution adjusts pH to 7.5, and the neutral proteinase that 0.3kg is then added is digested, and protects during enzymatic hydrolysis
It holds and stirs and maintain pH value of solution 7.5 ± 0.1 by the way that 3.0mol/L sodium hydroxide is added, digested at a temperature of 53 DEG C under ultrasound condition
4h；
(4) it extracts separation: enzymolysis liquid being warming up to 95 DEG C under stirring condition and keeps the temperature 2h, extracting solution is obtained, then uses
Centrifuge 4000rpm revolving speed by extracting solution slag charge and liquid separate, collect clear filtrate；
(5) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 40%；
(6) dry: medicinal extract 8h is dried in vacuo under the conditions of temperature 60 C and pressure -0.098MPa；
(7) it pulverizes and sieves: smashing it through 60 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Embodiment 4
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 160L/h extracts 4h under conditions of pumping frequency 30Hz, extracting pressure 25MPa, 40 DEG C of temperature, then
It is separated under the conditions of level-one separating pressure 10MPa, 40 DEG C of temperature and the second-order separation pressure 6MPa, 28 DEG C of temperature, collects oil
Rouge and degreasing ganoderma spove powder；
(3) it digests: 50kg degreasing ganoderma spove powder and 330kg pure water is mixed, be added under stirring condition
2.5mol/L sodium hydroxide solution, adjust pH to 7.0, then be added 0.3kg cellulase and 0.3kg compound protease into
Row digests, and is kept stirring during enzymatic hydrolysis and maintains pH value of solution 7.0 ± 0.1, ultrasound condition by the way that 2.5mol/L sodium hydroxide is added
Under digest 6h at a temperature of 48 DEG C；
(4) it extracts separation: enzymolysis liquid being warming up to 95 DEG C under stirring condition and keeps the temperature 2h, extracting solution is obtained, then uses
Centrifuge 5000rpm revolving speed by extracting solution slag charge and liquid separate, collect clear filtrate；
(5) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 45%；
(6) dry: medicinal extract 10h is dried in vacuo under the conditions of temperature 60 C and pressure -0.098MPa；
(7) it pulverizes and sieves: smashing it through 80 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Embodiment 5
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 140L/h extracts 5h under conditions of pumping frequency 25Hz, extracting pressure 18MPa, temperature 50 C, then
It is separated under the conditions of level-one separating pressure 8MPa, temperature 50 C and the second-order separation pressure 5MPa, 35 DEG C of temperature, collects grease
With degreasing ganoderma spove powder；
(3) it digests: 50kg degreasing ganoderma spove powder and 290kg pure water is mixed, be added under stirring condition
2.5mol/L potassium hydroxide solution adjusts pH to 7.8, and the compound protease that 0.5kg is then added is digested, and protects during enzymatic hydrolysis
It holds and stirs and maintain pH value of solution 7.8 ± 0.1 by the way that 2.5mol/L potassium hydroxide is added, digested at a temperature of 53 DEG C under ultrasound condition
4h；
(4) it extracts separation: enzymolysis liquid being warming up to 95 DEG C under stirring condition and keeps the temperature 2h, extracting solution is obtained, then uses
Centrifuge 6000rpm revolving speed by extracting solution slag charge and liquid separate, collect clear filtrate；
(5) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 35%；
(6) dry: medicinal extract 10h is dried in vacuo under the conditions of temperature 60 C and pressure -0.098MPa；
(7) it pulverizes and sieves: smashing it through 80 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Embodiment 6
A kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, comprising the following steps:
(1) broken wall: 100kg lucidum spore powder is fitted into vibration type ultramicro disintegrator, opens cooling water switch, to be cooled
After constant to 3-5 DEG C of water temperature, broken wall 30min makes sporoderm-broken rate reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall is put into supercritical carbon dioxide extracting kettle, is passed through overcritical dioxy
Change carbon flow body, flow 120L/h extracts 4h under conditions of pumping frequency 27.5Hz, extracting pressure 21MPa, temperature 45 C, so
It is separated, is collected under the conditions of level-one separating pressure 9MPa, temperature 45 C and the second-order separation pressure 5.5MPa, 31 DEG C of temperature afterwards
Grease and degreasing ganoderma spove powder；
(3) it extracts separation: degreasing ganoderma spove powder being mentioned into 2h in 95 DEG C of water, obtains extracting solution, then using centrifugation
Machine 5000rpm revolving speed by extracting solution slag charge and liquid separate, collect clear filtrate；
(4) be concentrated: clear filtrate is concentrated through economic benefits and social benefits vacuum energy-saving concentrator, effect a vacuum degree -0.06MPa, temperature 70
DEG C, two effect vacuum degree -0.08MPa, temperature 50 C are condensed into the medicinal extract of solid content 45%；
(5) dry: medicinal extract 12h is dried in vacuo under the conditions of 65 DEG C of temperature and pressure -0.084MPa；
(6) it pulverizes and sieves: smashing it through 80 mesh screens, obtain lucid ganoderma spore powder polysaccharide peptide powder.
Test example ganoderma polysaccharide peptide content detection
Ganoderma polysaccharide peptide content detection is carried out using high productivity computing method, using Waters1525 type high-efficient liquid phase color
Spectrum；Waters2487 UV detector；7.5 × 300mm of chromatographic column TSK-GEL G4000PWXL；Mobile phase: ultrapure water；Stream
Speed: 1.0mL/min；Column temperature: 35 DEG C；10 μ L of sample volume；Detection wavelength λ=280nm.
Specification Curve of Increasing
Dry peptidoglycan reference substance 10.0mg (purity >=98%, national Juncao Industry Technology Center) is accurately weighed, in
With water dissolution and constant volume in 10mL volumetric flask, the peptidoglycan reference substance stock solution of concentration 1.0mg/mL is obtained；Peptidoglycan is pipetted respectively
Reference substance stock solution is appropriate, is diluted with water and is settled to 2mL, and worth concentration is respectively the mark of 100,200,300,400,500 μ g/mL
Quasi- working solution, with reference substance concentration (μ g/mL) for abscissa, peak area is ordinate, draws standard curve.
Sample solution is prepared
The ganoderma polysaccharide peptide powder 50mg for weighing embodiment 1-6 preparation respectively, is dissolved in water and is settled to 5.0mL；Filtering,
Filtrate 1mL is taken, 3.0mL dehydrated alcohol is added and is mixed.It is centrifuged 10min in 10000rpm, the dissolution of precipitating water is settled to
5.0mL crosses 0.22 μm of filter membrane, it is spare to obtain sample filtrate.
Chromatography
It takes in 10 μ L sample solution injecting chromatographs and runs 10min, with peptidoglycan reference substance concentration (μ g/ml) for horizontal seat
Mark, peak area is ordinate, draws standard curve, calculates ganoderma lucidum polysaccharide peptide concentration in sample using the curve.
It is calculated by the following formula ganoderma lucidum polysaccharide peptide content, is as a result listed in Table 1.
Ganoderma lucidum polysaccharide peptide content in the ganoderma polysaccharide peptide powder of 1 embodiment 1-6 of table preparation
| Number | Polysaccharide peptide yield, % | Sample polysaccharide peptide content, % |
| Embodiment 1 | 9.24 | 35.75 |
| Embodiment 2 | 8.76 | 31.61 |
| Embodiment 3 | 9.01 | 32.33 |
| Embodiment 4 | 9.50 | 36.11 |
| Embodiment 5 | 8.97 | 34.23 |
| Embodiment 6 | 0.21 | 1.95 |
As shown in Table 1, ganoderma polysaccharide peptide prepared by the present invention, glycopeptide yield is in 8.76%-9.50%, wherein ganoderma lucidum polysaccharide
Peptide content is 31.61%-36.11%, and glycopeptide yield and content's index are obtained through refining standby lucidum spore powder significantly better than simple water and obtained
Peptidoglycan.
Claims (10)
1. a kind of preparation method of lucid ganoderma spore powder polysaccharide peptide, which comprises the following steps:
(1) broken wall: low-temperature physics broken wall treatment is carried out to lucidum spore powder, sporoderm-broken rate is made to reach 95% or more；
(2) degreasing: the lucidum spore powder after broken wall being put into supercritical carbon dioxide extraction apparatus and is extracted, and is then carried out
It separates twice, collects grease and degreasing ganoderma spove powder；
(3) it digests: degreasing ganoderma spove powder is mixed with pure water, add aqueous slkali to adjust pH to 5-10, enzyme is then added
It is digested；
(4) it extracts filtering: enzymolysis liquid being warming up to 90-100 DEG C under stirring condition and keeps the temperature 1-2h, extracting solution is obtained, then divides
From collection clear filtrate；
(5) it is concentrated: concentrating the filtrate to the medicinal extract of solid content 35-55%；
(6) dry: vacuum drying medicinal extract；
(7) it pulverizes and sieves: smashing it through 60-80 mesh screen, obtain lucid ganoderma spore powder polysaccharide peptide powder.
2. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the low-temperature physics is broken
Wall is carried out using vibration type ultramicro disintegrator, broken time 25-35min.
3. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the extraction equipment
Pumping frequency is 25-30Hz, flow >=120L/h of supercritical carbon dioxide, extracting pressure 18-25MPa, extraction temperature 40-50
DEG C, extraction time is 4-5 hours.
4. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that described to separate twice
Level-one separating pressure is 8-10MPa, and separation temperature is 40-50 DEG C；The second-order separation pressure is 5-6MPa, separation temperature 28-35
℃。
5. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the aqueous slkali is hydrogen
One of sodium oxide molybdena, potassium hydroxide, calcium hydroxide and ammonium hydroxide.
6. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that aqueous slkali is added and adjusts
PH to 7.0-8.5.
7. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the enzyme is cellulose
One of enzyme, compound protease, alkali protease and neutral proteinase are a variety of；The additional amount of the enzyme is the spirit of degreasing broken wall
The 0.1-2% of Ganoderma lucidum spore powder weight.
8. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the temperature of the enzymatic hydrolysis
It is 40-55 DEG C, the time of enzymatic hydrolysis is 2-12h.
9. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that carried out during enzymatic hydrolysis
Ultrasonic treatment, the frequency of ultrasonic treatment are 20-40KHz, time 4-6h.
10. the preparation method of lucid ganoderma spore powder polysaccharide peptide according to claim 1, which is characterized in that the concentration uses
Economic benefits and social benefits vacuum energy-saving concentrator carries out；One effect vacuum degree of economic benefits and social benefits vacuum energy-saving concentrator is -0.06Mpa, temperature 70-
80℃；Two effect vacuum degrees are -0.08MPa, and temperature is 50-60 DEG C.
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| CN113912750B (en) * | 2021-11-11 | 2022-11-11 | 开平健之源保健食品有限公司 | Method for extracting ganoderma lucidum fruiting body polysaccharide through fermentation pretreatment |
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