CN108341890A - A kind of extracting method of ganoderma lucidum polysaccharide - Google Patents
A kind of extracting method of ganoderma lucidum polysaccharide Download PDFInfo
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Abstract
The invention discloses a kind of extracting methods of ganoderma lucidum polysaccharide, and ganoderma lucidum fruitbody first crushed 80 mesh sieve, place into and complete explosion in steam blasting device, by solid-to-liquid ratio 1:30 ~ 50 DEG C are heated to after 10 ~ 20 addition water immersion 1.5h to be ultrasonically treated, adjust feed temperature, pH value, protease and cellulase, heat up enzyme deactivation work, centrifugation, resin decolorization, de- albumen, filtering, reduced pressure, vacuum dehydrating at lower temperature after enzymolysis, obtain the ganoderma lucidum polysaccharide solid powder of high activity, high-purity, improve polysaccharide extract rate and production efficiency, and the space structure and its bioactivity of reduction of polysaccharide will not be destroyed, it is more suitable for industrialized production.
Description
Technical field
The invention belongs to technical field of polysaccharide extraction, and in particular to a kind of extracting method of ganoderma lucidum polysaccharide.
Background technology
Ganoderma lucidum（Ganoderma lucidum）It is the high macro-fungi of a kind of nutrition, health value.Since ancient times,
Ganoderma lucidum is regarded as lucky, rich and honour, fine and long-lived symbol, has the title of " celestial grass " and " seocho ", chinese tradition medicine long
Since phase a direct-view for nourishing and fit keeping function, consolidate the precious Chinese herbal medicine of righting.If fungi is by directly edible typically up to less than treatment
The effect of, active ingredient is usually only passed through extracting and developing and is purified, and could improve effect, play certain pharmacological action.Greatly
Quantity research report display：Ganoderma lucidum polysaccharide is ganoderma lucidum primary bioactive components, the various drug effects of ganoderma lucidum and the activity of ganoderma lucidum polysaccharide
It is closely related with content, and ganoderma lucidum polysaccharide has become one of the most important index for weighing lucid ganoderma product quality quality.
In active material preparation process, extraction is the first step and a most key step, how by active ingredient most
Big releasing for degree has very important significance.Ganoderma lucidum polysaccharide traditional extraction process mainly has Hot water extraction, alkaline process
And acid system.Different from other edible mushrooms, ganoderma lucidum fruitbody degree of lignification is high, complicated, polysaccharide generally with cellulose, wooden
Element is combined by being physically or chemically bonded, and ganoderma lucidum fruitbody is made to have stronger hardness, and ganoderma lucidum polysaccharide is present in son in fact
It is difficult to come out Polyose extraction using general method in the cell wall of body.From current result of study can be seen that merely into
The extraction of row water solubility ganoderma lucidum polysaccharide, recovery rate only has 3% or so, and remaining most polysaccharide is all trapped in residue,
If directly abandoned, that is, result in waste of resources, and pollute the environment.Traditional water, alkali, acidleach formulation ganoderma lucidum polysaccharide carry
Take the time long, energy consumption is big, and Extracting temperature is high, and long-time high temperature makes ganoderma lucidum polysaccharide there is a possibility that ganoderma lucidum polysaccharide is degraded
Extraction be extremely difficult to ideal effect, these all greatly limit the development and utilization of ganoderma lucidum polysaccharide.To overcome traditional water to extract
There is the new extractive technique and method such as ultrasound, microwave auxiliary extraction, super-pressure, enzyme process, still in recent years in the deficiency of method
The extraction effect of these methods is also very undesirable.
To overcome the defect of above method, the present invention using steam explosion come clear up the lignin in ganoderma lucidum fruitbody and
Cellulose makes ganoderma lucidum polysaccharide more release, and recycles ultrasound assisted extraction, macroreticular resin decolorization, finally obtains
Polysaccharide of the retention rate 85% or more.
Invention content
In order to solve the above technical problems, the present invention provides a kind of extracting methods of the higher ganoderma lucidum polysaccharide of yield.
The extracting method of ganoderma lucidum polysaccharide of the present invention is realized by following technical solutions：
A kind of extracting method of ganoderma lucidum polysaccharide, includes the following steps：
（1）It crushes：Dry ganoderma lucidum fruitbody is crushed through biomass pulverizer, 80 mesh sieve is crossed, obtains glossy ganoderma powder；
（2）Steam blasting：100g glossy ganoderma powders are put into steam blasting device and complete explosion, obtain explosion glossy ganoderma powder；
（3）It is ultrasonically treated：Water is added according to solid-to-liquid ratio in quick-fried glossy ganoderma powder and impregnates 1.5h, and is aided with supersound process at 30 ~ 50 DEG C,
Obtain material liquid mixture；
（4）Proteolysis：Material liquid mixture is added in retort, temperature, the pH value of retort are controlled, protease is put into, holds
1 ~ 2h of continuous stirring carries out proteolysis；
（5）Fiber digests：Again control retort in feed liquid temperature and pH value, then put into cellulase, persistently stir 1 ~
2h carries out fiber enzymolysis, obtains enzymolysis liquid；
（6）Enzyme deactivation is lived：Feed liquid is warming up to 85 DEG C, keeps 15min, obtains enzyme deactivation work liquid；
（7）Decoloration：Enzyme deactivation is lived after liquid centrifugation, and recycling feed liquid is in bleacher, and circulating water is to 40 ~ 60 DEG C, input nonpolarity
Macroporous absorbent resin, decolourize 1h under stirring, obtains decoloration feed liquid；
（8）It is concentrated under reduced pressure：Feed liquid of decolourizing filtering, low-temperature reduced-pressure are concentrated into the 1/3 of original volume, obtain ganoderma lucidum polysaccharide concentrate；
（9）It is dry：Ganoderma lucidum polysaccharide concentrate obtains ganoderma lucidum polysaccharide solid powder after vacuum dehydrating at lower temperature.
Further, the steam blasting pressure is 1.5MPa-2.0MPa, temperature is 100 DEG C -200 DEG C, the time is
2min-5min。
Further, the solid-to-liquid ratio is 1:10~20.
Further, the ultrasonic power is 3000 ~ 5000W, and the processing time is 30min.
Further, the proteolysis temperature is 45 ~ 60 DEG C, and pH value 6.0 ~ 8.0, albumen enzyme dosage accounts for glossy ganoderma powder weight
Than 0.5 ~ 1.5%.
Further, the fiber hydrolysis temperature is 45 ~ 60 DEG C, and pH value 3.0 ~ 6.0, cellulase dosage accounts for glossy ganoderma powder weight
Measure ratio 1.0 ~ 3.0%.
Further, the nonpolar macroporous adsorption resin dosage accounts for centrifugation feed liquid weight ratio 5.0 ~ 10.0%.
Further, the low-temperature reduced-pressure thickening temperature is 50 ~ 80 DEG C.
Further, the vacuum dehydrating at lower temperature temperature is 60 ~ 90 DEG C.
The beneficial effects of the present invention are：
Compared with prior art, beneficial effects of the present invention are：
（1）The present invention is handled ganoderma lucidum fruitbody using steam explosion, can be effectively by cellulose therein and lignin
Resolution or fracture, make more ganoderma lucidum polysaccharide release, substantially increase the extraction efficiency of polysaccharide.
（2）The present invention is combined the release, diffusion and dissolving for strengthening intracellular organic matter with steam blasting using supersound process, more
Conducive to the dissolution of active polysaccharide, the usage amount of enzyme, easy to operate, mild condition are reduced.
（3）Using cellulase, the stepped combined enzymolysis of protease, it is sufficiently destroyed substrate eucaryotic cell structure, is discharged to greatest extent
Polysaccharide, while keeping the bioactivity of ganoderma lucidum polysaccharide, using nonpolar macroporous adsorption resin to extracting liquid decoloration, de- albumen at
Reason, percent of decolourization and polysaccharide yield are high, will not destroy polysaccharide structures.
（4）Ganoderma lucidum polysaccharide is extracted using the present invention, there is the features such as yield is high, easy to operate, for excavating spirit
The potential value of sesame has very important meaning.
Specific implementation mode
The present invention is further described with reference to specific embodiment, so that those skilled in the art knows more about
The present invention, but be not intended to limit the present invention.
Embodiment 1
（1）It crushes：Dry ganoderma lucidum fruitbody is crushed through biomass pulverizer, 80 mesh sieve is crossed, obtains glossy ganoderma powder；
（2）Steam blasting：100g glossy ganoderma powders are put into steam blasting device, burst pressure 1.5MPa, temperature are 150 DEG C, tie up
Explosion 2min is held, then the pressure in reactor is reduced into 1 standard atmospheric pressure, after completing explosion, obtains explosion glossy ganoderma powder；
（3）It impregnates：Explosion glossy ganoderma powder is added in retort, by solid-to-liquid ratio 1:15, which are added water, impregnates 1.5h, obtains material liquid mixture；
（4）It is ultrasonically treated：Material liquid mixture carries out 30min supersound process, ultrasonic power 4000W at 40 DEG C；
（5）Proteolysis：52 DEG C of temperature, the pH value 7.0 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
1.0% protease digests 2h under stirring；
（6）Fiber digests：52 DEG C of temperature, the pH value 4.5 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
2.0% cellulase digests 2h under stirring；
（7）Enzyme deactivation is lived：Feed liquid is warming up to 85 DEG C, keeps 15min, obtains enzymolysis liquid；
（8）Decoloration：After enzymolysis liquid centrifugation, recycling feed liquid is in bleacher, and circulating water is to 50 DEG C, input centrifugation feed liquid 7.5%
Nonpolar macroporous adsorption resin（Resin provides producer：ROHM AND HAAS, resin model：AMBERLITE XAD16 macroporous absorption trees
Fat）, decolourize 1h under stirring, obtains decoloration feed liquid；
（9）It is concentrated under reduced pressure：Feed liquid of decolourizing filtering, is concentrated under reduced pressure into the 1/3 of original volume at 75 DEG C, obtains ganoderma lucidum polysaccharide concentration
Liquid；
（10）It is dry：Ganoderma lucidum polysaccharide concentrate after vacuum dehydrating at lower temperature, obtains ganoderma lucidum polysaccharide solid powder at 90 DEG C.
Embodiment 2
（1）It crushes：Dry ganoderma lucidum fruitbody is crushed through biomass pulverizer, 80 mesh sieve is crossed, obtains glossy ganoderma powder；
（2）Steam blasting：100g glossy ganoderma powders are put into steam blasting device, steam blasting pressure 1.5MPa, temperature 150
DEG C, maintain explosion 5min, then the pressure in reactor is reduced into 1 standard atmospheric pressure, after completing explosion, obtains explosion glossy ganoderma powder；
（3）It impregnates：Explosion glossy ganoderma powder is added in retort, by solid-to-liquid ratio 1:10, which are added water, impregnates 1.5h, obtains material liquid mixture；
（4）It is ultrasonically treated：Material liquid mixture carries out 30min supersound process, ultrasonic power 5000W at 40 DEG C；
（5）Proteolysis：52 DEG C of temperature, the pH value 7.0 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
1.0% protease digests 2h under stirring；
（6）Fiber digests：52 DEG C of temperature, the pH value 4.5 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
2.0% cellulase digests 2h under stirring；
（7）Enzyme deactivation is lived：Feed liquid is warming up to 85 DEG C, keeps 15min, obtains enzymolysis liquid；
（8）Decoloration：After enzymolysis liquid centrifugation, recycling feed liquid is in bleacher, and circulating water is to 55 DEG C, input centrifugation feed liquid 5.0%
Nonpolar macroporous adsorption resin, decolourize 1h under stirring, obtains decoloration feed liquid；
（9）It is concentrated under reduced pressure：Feed liquid of decolourizing filtering, is concentrated under reduced pressure into the 1/3 of original volume at 70 DEG C, obtains ganoderma lucidum polysaccharide concentration
Liquid；
（10）It is dry：Ganoderma lucidum polysaccharide concentrate after vacuum dehydrating at lower temperature, obtains ganoderma lucidum polysaccharide solid powder at 90 DEG C.
Embodiment 3
（1）It crushes：Dry ganoderma lucidum fruitbody is crushed through biomass pulverizer, 80 mesh sieve is crossed, obtains glossy ganoderma powder；
（2）Steam blasting：100g glossy ganoderma powders are put into steam blasting device, steam blasting pressure 2.0MPa, temperature 200
DEG C, maintain explosion 3min, then the pressure in reactor is reduced into 1 standard atmospheric pressure, after completing explosion, obtains explosion glossy ganoderma powder；
（3）It impregnates：Explosion glossy ganoderma powder is added in retort, by solid-to-liquid ratio 1:20, which are added water, impregnates 1.5h, obtains material liquid mixture；
（4）It is ultrasonically treated：Material liquid mixture carries out 30min supersound process, ultrasonic power 3000W at 50 DEG C；
（5）Proteolysis：52 DEG C of temperature, the pH value 7.0 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
1.0% protease digests 2h under stirring；
（6）Fiber digests：52 DEG C of temperature, the pH value 4.5 for controlling feed liquid in retort put into glossy ganoderma powder weight ratio into retort
2.0% cellulase digests 2h under stirring；
（7）Enzyme deactivation is lived：Feed liquid is warming up to 85 DEG C, keeps 15min, obtains enzymolysis liquid；
（8）Decoloration：After enzymolysis liquid centrifugation, recycling feed liquid is in bleacher, and circulating water is to 55 DEG C, input centrifugation feed liquid
10.0% nonpolar macroporous adsorption resin, decolourize 1h under stirring, obtains decoloration feed liquid；
（9）It is concentrated under reduced pressure：Feed liquid of decolourizing filtering, is concentrated under reduced pressure into the 1/3 of original volume at 80 DEG C, obtains ganoderma lucidum polysaccharide concentration
Liquid；
（10）It is dry：Ganoderma lucidum polysaccharide concentrate after vacuum dehydrating at lower temperature, obtains ganoderma lucidum polysaccharide solid powder at 90 DEG C.
Comparative example 1
Extraction step is same as Example 1, only cancels Steam explosion treatment.
Comparative example 2
Extraction step is same as Example 1, only cancels being ultrasonically treated.
Comparative example 3
Extraction step is same as Example 1, only cancels Steam explosion treatment and is ultrasonically treated.
Test example
Embodiment 1 ~ 3 is detected with the polyoses content extracted in comparative example 1 ~ 3, the extraction effect of more different extracting modes
Fruit.
1. the measurement of ganoderma polyoses content
1.1 assay method
The ganoderma lucidum polysaccharide that Different Extraction Method obtains, content assaying method refer to the phend-sulphuric acid of the reports such as Dubois, together
When, is made suitably modified, and glucose is as standard items.
The drafting of 1.2 glucose standard curves
(1) glucose standards solution
Accurate 100 mg of glucose for weighing constant weight, deionized water dissolving are settled to 100 m L.Solution 10ml is taken out to add
Deionized water is settled to 100 ml, obtains the glucose standards solution of 100 μ g/m L.
(2) 5% phenol solution
Phenol accurately is weighed, with deionized water dissolving, is configured to mass volume ratio 5%（g/m L）Solution shading it is spare.
(3) the drafting of standard curve
6 test tubes, number 1,2,3,4,5,6 are taken to be separately added into 100 Portugals μ g/mL of 0.3,0.6,0.9,1.2,1.5,1.8 ml
Grape Standard for Sugars liquid, respectively plus deionized water is mended to 2 ml, then is separately added into 1mL5% phenol solutions, on eddy mixer fully
Mixing, finally adds the 5ml concentrated sulfuric acids, after fully shaking up chromogenic reaction 20min under room temperature, with ultraviolet point of UV-1700 types
Light photometric determination respectively numbers the light absorption value at 490nm.Using concentration of glucose as abscissa, light absorption value A490 is at 490 nm
Ordinate draws concentration of glucose-absorbance value regression curve.
(4) sugared content in phend-sulphuric acid determination sample
The polysaccharide solution of debita spissitudo is prepared, it is accurate to draw polysaccharide solution 2mL, add 5% phenol solution 1.0mL mixings, dense sulphur
Sour 5.0mL is mixed well, and colour developing surveys its absorbance value, according to the extinction of glucose standard curve and polysaccharide at 490nm wavelength
Value calculates ganoderma lucidum polysaccharide yield.
1.3 polysaccharide yield calculation formula
Ganoderma lucidum polysaccharide yield Y (%)=(C × V0×V1/V2)/W×10-6×100%
W is the weight of ganoderma lucidum fruitbody powder（g）；C is is surveyed liquid polysaccharide concentration（μg/m L）；V0 is developing solution total volume
（mL）；V1 is the volume of total ganoderma lucidum polysaccharide solution（mL）；V2 is the volume of ganoderma lucidum polysaccharide sample liquid added in development step（m
L）.
2. result
The different group ganoderma polyoses contents of table 1 measure
。
As shown in Table 1,90% or more, comparative example 1 takes the ganoderma lucidum polysaccharide yield that prepared by embodiment 1 ~ 3 compared to embodiment 1
Disappear Steam explosion treatment, and ganoderma lucidum polysaccharide yield is 75.5% significantly lower than embodiment 1；Comparative example 2 cancels ultrasound compared to embodiment 1
Processing, ganoderma lucidum polysaccharide yield are 80.6% significantly lower than embodiment 1 but are higher than comparative example 1；Comparative example 3 is cancelled compared to embodiment 1 steams
Vapour explosion treatment and supersound process, ganoderma lucidum polysaccharide yield are 62.3% significantly lower than comparative example 1.Illustrate only in steam blasting and
Under the collaboration processing of supersound process, ganoderma lucidum polysaccharide yield could be improved.
The above, only presently preferred embodiments of the present invention, not to the present invention in any form with substantial limitation,
It should be pointed out that for those skilled in the art, under the premise of not departing from the method for the present invention, can also make
Several improvement and supplement, these are improved and supplement also should be regarded as protection scope of the present invention.All those skilled in the art,
Without departing from the spirit and scope of the present invention, when made using disclosed above technology contents it is a little more
Dynamic, modification and the equivalent variations developed, are the equivalent embodiment of the present invention；Meanwhile all substantial technologicals pair according to the present invention
The variation, modification and evolution of any equivalent variations made by above-described embodiment, still fall within the range of technical scheme of the present invention
It is interior.
Claims (10)
1. a kind of extracting method of ganoderma lucidum polysaccharide, which is characterized in that this method comprises the following steps：
（1）It crushes：Dry ganoderma lucidum fruitbody is crushed through biomass pulverizer, 80 mesh sieve is crossed, obtains glossy ganoderma powder；
（2）Steam blasting：100g glossy ganoderma powders are put into steam blasting device and complete explosion, obtain explosion glossy ganoderma powder；
（3）It is ultrasonically treated：Water is added according to solid-to-liquid ratio in quick-fried glossy ganoderma powder and impregnates 1.5h, and is aided with supersound process at 30 ~ 50 DEG C,
Obtain material liquid mixture；
（4）Proteolysis：Material liquid mixture is added in retort, temperature, the pH value of retort are controlled, protease is put into, holds
1 ~ 2h of continuous stirring carries out proteolysis；
（5）Fiber digests：Again control retort in feed liquid temperature and pH value, then put into cellulase, persistently stir 1 ~
2h carries out fiber enzymolysis, obtains enzymolysis liquid；
（6）Enzyme deactivation is lived：Feed liquid is warming up to 85 DEG C, keeps 15min, obtains enzyme deactivation work liquid；
（7）Decoloration：Enzyme deactivation is lived after liquid centrifugation, and recycling feed liquid is in bleacher, and circulating water is to 40 ~ 60 DEG C, input nonpolarity
Macroporous absorbent resin, decolourize 1h under stirring, obtains decoloration feed liquid；
（8）It is concentrated under reduced pressure：Feed liquid of decolourizing filtering, low-temperature reduced-pressure are concentrated into the 1/3 of original volume, obtain ganoderma lucidum polysaccharide concentrate；
（9）It is dry：Ganoderma lucidum polysaccharide concentrate obtains ganoderma lucidum polysaccharide solid powder after vacuum dehydrating at lower temperature.
2. extracting method according to claim 1, which is characterized in that the steam blasting pressure be 1.5MPa-2.0MPa,
Temperature is 100 DEG C -200 DEG C, time 2min-5min.
3. extracting method according to claim 1, which is characterized in that the solid-to-liquid ratio is 1:10~20.
4. extracting method according to claim 1, which is characterized in that the ultrasonic power is 3000 ~ 5000W, the place
The reason time is 30min.
5. extracting method according to claim 1, which is characterized in that the proteolysis temperature is 45 ~ 60 DEG C, pH value
6.0 ~ 8.0, albumen enzyme dosage accounts for glossy ganoderma powder weight ratio 0.5 ~ 1.5%.
6. extracting method according to claim 1, which is characterized in that the fiber hydrolysis temperature is 45 ~ 60 DEG C, pH value
3.0 ~ 6.0, cellulase dosage accounts for glossy ganoderma powder weight ratio 1.0 ~ 3.0%.
7. extracting method according to claim 1, which is characterized in that the resin is nonpolar macroporous adsorption resin dosage
Account for centrifugation feed liquid weight ratio 5.0 ~ 10.0%.
8. extracting method according to claim 1, which is characterized in that the low-temperature reduced-pressure thickening temperature is 50 ~ 80 DEG C.
9. extracting method according to claim 1, which is characterized in that the vacuum dehydrating at lower temperature temperature is 60 ~ 90 DEG C.
10. a kind of according to ganoderma lucidum polysaccharide made from any means described in claim 1 ~ 9.
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