CN106220751A - A kind of method of ultrasonic assistant multiplex-enzyme extraction Armillariella mellea polysaccharide - Google Patents
A kind of method of ultrasonic assistant multiplex-enzyme extraction Armillariella mellea polysaccharide Download PDFInfo
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- CN106220751A CN106220751A CN201610800639.7A CN201610800639A CN106220751A CN 106220751 A CN106220751 A CN 106220751A CN 201610800639 A CN201610800639 A CN 201610800639A CN 106220751 A CN106220751 A CN 106220751A
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
A kind of method that the invention discloses ultrasonic assistant multiplex-enzyme extraction Armillariella mellea polysaccharide, belongs to the preparation field extracting polysaccharide.This preparation method comprises the following steps: Armillariella mellea is cleaned, dry, pulverize after cross 60 mesh sieves, obtain Armillariella mellea powder；Enzymolysis, 100 DEG C of enzyme denaturing, cooling is carried out according to different conditions；By said extracted liquid again after the supersound process of different condition, centrifugal, supernatant is polysaccharide extraction liquid.By single factor experiment, finally determine the extraction the suitableeest liquid ratio of Armillariella mellea polysaccharide, enzymolysis time, hydrolysis temperature, compound enzyme ratio, enzyme concentration, ultrasonic time, ultrasonic power.The present invention have studied the impact on Armillariella mellea polysaccharide extract rate of the differential responses condition, optimizes and draws the optimal condition extracting Armillariella mellea polysaccharide.
Description
Technical field
A kind of method that the present invention relates to ultrasonic assistant multiplex-enzyme extraction Armillariella mellea polysaccharide, belongs to the preparation extracting polysaccharide
Field.
Background technology
Armillariella mellea has another name called Armillaria mellea, and it is under the jurisdiction of mycota, is the sporophore of Tricholomataceae fungus Armillaria mellea, is prevalent in temperature
Band area.Armillariella mellea tasty mouthfeel, rich in abundant nutrient substance, as protein, carbohydrate, vitamin, Yi Jixin, ferrum,
The mineral such as magnesium, therefore can develop and have functional Armillariella mellea food.Additionally, Armillariella mellea or a kind of food and medicament dual-purpose fungus.
Armillariella mellea polysaccharide separates from Armillariella mellea mycelium, sporophore and tunning, is that one can control carefully
The Differentiation of born of the same parents and the active polysaccharide of regulation cell growth aging, it also has radioprotective, promotes hemopoietic, suppression tumor simultaneously
The pharmacological actions such as growth, regulation immunity.Additionally, Armillariella mellea polysaccharide also has certain oxidation resistance, may be used for preparation and have
The natural active matter of antioxidant activity.
Armillariella mellea Polyose extraction can use hot water, diluted acid, diluted alkaline as digestion agent, it is possible to uses supercritical ultrasonics technology auxiliary enzyme process
Extract.Extraction is the traditional method extracting Armillariella mellea polysaccharide, but time length, efficiency are low, energy consumption is big, this external diluted acid, diluted alkaline bar
Under part, easily make the fracture of polysaccharide generation glycosidic bond, par-tial polysaccharide occurs hydrolysis to make the extraction ratio of polysaccharide reduce, therefore polysaccharide
Extraction ratio is the highest.Ultrasonic assistant enzyme process extracts polysaccharide simple and fast, save time low consumption, and the industrialization to Armillariella mellea polysaccharide is extracted
And production has very important realistic meaning.
About research existing a lot of report at home and abroad of Armillariella mellea polysaccharide, relate to each sides such as biochemistry, pharmacology, clinic
Face, illustrates its structure, function and structure activity relationship, is of great significance the development tool of Chinese medicine new medicine.
Li Yan equality research shows, Armillariella mellea polysaccharide, by stimulating medullary cell to make juvenile cell increase, is divided into ripe thin
Born of the same parents, cause segmented cell to increase, and protect medulla hematopoietic system, and the damage to medullary cell of the antagonism cyclophosphamide, it acts on knot
Fruit is in dose-dependent relationship.But the research to these polysaccharide mainly comes from the Armillaria mellea fermented product of liquid culture with artificial
The Armillaria mellea mycelia extracting solution of solid culture, isolation identification and the most rarely seen report of Study on mechanism thereof to wildness hazel-mushroom polysaccharide
Road.
Summary of the invention
It is an object of the invention to provide a kind of method extracting Armillariella mellea polysaccharide, extracting method of the present invention utilizes ultrasound wave to combine
Compound enzyme extracts Armillariella mellea polysaccharide, is optimized by extraction conditions, and preparation-obtained Armillariella mellea polysaccharide properties is stable.
It is an object of the invention to be achieved through the following technical solutions:
A kind of method taking Armillariella mellea polysaccharide, uses ultrasound wave to combine compound enzyme to extract Armillariella mellea polysaccharide, comprises the following steps:
Armillariella mellea is cleaned, dry, pulverize, cross 60 mesh sieves, obtain Armillariella mellea powder；Enzymolysis is carried out according to different conditions, 100 DEG C of enzyme denaturing,
Cooling；By said extracted liquid again after the supersound process of different condition, centrifugal, supernatant is polysaccharide extraction liquid.
The present invention have studied the condition of complex enzyme zymohydrolysis, at different liquid ratios, enzymolysis time, hydrolysis temperature, compound enzyme
Impact under the conditions of ratio, enzyme concentration etc., on Armillariella mellea polysaccharide extract rate.According to result of the test, determine the horizontal combination of optimum.
Being drawn by single factor experiment, obtaining optimum process condition is: hydrolysis temperature 50 DEG C, ultrasonic power 360W, super
During sound wave time 20min, enzyme concentration 1.75%, compound enzyme ratio 2:1, enzymolysis time 140min, liquid ratio 30:1 (mL/g),
The Armillariella mellea polysaccharide extract rate obtained under the conditions of this is maximum.
Accompanying drawing explanation
The impact on Armillariella mellea polysaccharide extract rate of Fig. 1 liquid ratio；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 2 enzymolysis time；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 3 hydrolysis temperature；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 4 enzyme concentration；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 5 compound enzyme ratio；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 6 ultrasonic power；
The impact on Armillariella mellea polysaccharide extract rate of Fig. 7 ultrasonic time；
Detailed description of the invention
Below in conjunction with the accompanying drawings technical scheme is further described, but is not limited thereto, every to this
Inventive technique scheme is modified or equivalent, without deviating from the spirit and scope of technical solution of the present invention, all should contain
In protection scope of the present invention.
One, the preparation of Armillariella mellea powder
Dry after wildness hazel-mushroom is cleaned, cross 60 mesh sieves after pulverizing with pulverizer, obtain Armillariella mellea powder, standby.
Two, complex enzyme zymohydrolysis
Take the wildness hazel-mushroom sample 5g that particle diameter is 60 mesh, be configured to liquid ratio and be respectively the aqueous solution of 20:1, the most enzyme-added
(enzyme-added total amount is the 1% of substrate) (cellulase: papain=1:1), pH is transferred to analog value, hydrolysis temperature 45 DEG C, ph
Value 5, enzymolysis time 2h, with 100 DEG C of boiling water pot water-bath 10min, enzyme denaturing, be cooled to room temperature.It is respectively through ultrasonic power again
After the process 20min of 80w, with 4000r/min centrifuge 30min, take supernatant, i.e. obtain enzyme extraction polysaccharide clear liquid.Root
Corresponding test parameters is changed according to the single factor test investigated.
1, single factor experiment design
Investigate compound enzyme ratio (cellulase: papain) (3:1,2:1,1:1,1:2,1:3), enzyme concentration respectively
(accounting for the percentage ratio of substrate) (1.00,1.25,1.50,1.75,2.00%), liquid ratio (10:1,20:1,30:1,40:1,50:
1), hydrolysis temperature (40,45,50,55,60 DEG C), enzymolysis time (80,100,120,140,160min), ultrasonic power (240,
280,320,360,400W) and ultrasonic time (10,15,20,25,30min) etc. 7 factors shadow to Armillariella mellea polysaccharide extract rate
Ring.
Three, experiment of single factor
1, the liquid ratio impact on explanation polysaccharide extract rate
As it is shown in figure 1, when liquid ratio increases to 20:1 from 10:1, the extraction ratio of Armillariella mellea polysaccharide can significantly raise, from
16.68% increases to 38.55%；When liquid ratio continues to increase, and the extraction ratio of polysaccharide reduces on the contrary, therefore select 20:1 as hazel
The optimal liquid ratio of mushroom polysaccharide.
2, the enzymolysis time impact on Armillariella mellea polysaccharide extract rate
From figure 2 it can be seen that when enzymolysis time increases to 140min from 80min, the extraction ratio of Armillariella mellea polysaccharide is notable
Raise；But, when enzymolysis time increases to 160min, the extraction ratio of Armillariella mellea polysaccharide is remarkably decreased again, therefore selects 140min to make
For extracting the optimal enzymolysis time of Armillariella mellea polysaccharide.
3, the hydrolysis temperature impact on Armillariella mellea polysaccharide extract rate
From figure 3, it can be seen that when temperature is in time being increased to 50 DEG C for 40 DEG C, the extraction ratio of Armillariella mellea polysaccharide increases from 31.65%
It is added to 34.16%,；When temperature continues to raise, and the extraction ratio of Armillariella mellea polysaccharide can be remarkably decreased, therefore select 50 DEG C as extracting hazel
The optimal hydrolysis temperature of mushroom polysaccharide.
4, the enzyme concentration impact on Armillariella mellea polysaccharide extract rate
Figure 4, it is seen that when enzyme concentration increases to 1.75% from 1.00%, the extraction ratio of Armillariella mellea polysaccharide from
25.22% increases to 37.44%,；When enzyme concentration continues to increase to 2.00%, polysaccharide extract rate is reduced to 35.41, therefore selects
1.75% makees to extract the optimal enzyme concentration of Armillariella mellea polysaccharide.
5, the compound enzyme ratio impact on Armillariella mellea polysaccharide extract rate
From figure 5 it can be seen that when compound enzyme ratio becomes 2:1 from 3:1, polysaccharide extract rate increases to from 34.58
40.46, Armillariella mellea polysaccharide extract rate increases；When the ratio of compound enzyme gradually becomes 1:3 from 2:1, polysaccharide extract rate is reduced to
31.54, Armillariella mellea polysaccharide extract rate reduces, and therefore selects 2:1 as the best complex enzyme ratio extracting Armillariella mellea polysaccharide.
6, the ultrasonic power impact on Armillariella mellea polysaccharide extract rate
From fig. 6 it can be seen that when ultrasonic power increases to 360W from 240W, polysaccharide extract rate increases from 28.04%
To 31.38%, Armillariella mellea polysaccharide extract rate increases；But if ultrasonic power continues to increase, polysaccharide extract rate reduces, and therefore selects
360W is as the optimal ultrasonic power extracting Armillariella mellea polysaccharide.
7, the ultrasonic time impact on Armillariella mellea polysaccharide extract rate
It can be seen from figure 7 that when ultrasonic time increases to 20min from 10min, polysaccharide extract rate increases from 29.76%
Being added to 32.17%, the extraction ratio of Armillariella mellea polysaccharide raises；And when ultrasonic time continues to increase, polysaccharide extract rate declines on the contrary, because of
This selects 20min as the optimal ultrasonic time extracting Armillariella mellea polysaccharide.
According to monofactorial result, determine the best of breed of Armillariella mellea Polyose extraction, draw the optimum extraction work of Armillariella mellea polysaccharide
Skill: enzyme concentration 1.75%, compound enzyme ratio 2:1, enzymolysis time 140min, liquid ratio 30:1 (mL/g), hydrolysis temperature 50 DEG C, super
Acoustical power 360W, ultrasonic time 20min.In order to verify the accuracy of this test, polysaccharide extract rate is carried out proving test, flat
Row test 3 times, the meansigma methods of extraction ratio is 39.57%.
Claims (9)
1. the method for a ultrasonic assistant multiplex-enzyme extraction Armillariella mellea polysaccharide, it is characterised in that described method step is as follows: step
One: wildness hazel-mushroom is cleaned dried and crushed machine and pulverizes, make Armillariella mellea powder；Step 2: by water from Armillariella mellea powder according to different
Liquid ratio (10:1,20:1,30:1,40:1,50:1) mixes, wherein: compound enzyme ratio (cellulase: papain) (3:
1,2:1,1:1,1:2,1:3), enzyme concentration (accounting for the percentage ratio of substrate) (1.00,1.25,1.50,1.75,2.00%), enzymolysis temperature
Degree (40,45,50,55,60 DEG C), enzymolysis time (80,100,120,140,160min), 100 DEG C of enzyme denaturing, cooling；Step 3: will
Solution supersound process after above-mentioned process, ultrasonic power (240,280,320,360,400W) and ultrasonic time (10,
15,20,25,30min), be centrifuged afterwards, supernatant is Armillariella mellea polysaccharide extraction liquid.
2. according to the preparation method described in claim 1, it is characterised in that the Armillariella mellea powder described in step one should cross 60 mesh sieves,
Standby.
3. according to the preparation method described in claim 1, it is characterised in that the liquid ratio of the solution described in step 2 is 20:1.
4. according to the preparation method described in claim 1, it is characterised in that the compound enzyme ratio (cellulose described in step 2
Enzyme: papain) it is 2:1.
5. according to the preparation method described in claim 1, it is characterised in that the enzyme concentration of the solution described in step 2 (accounts for substrate
Percentage ratio) be 1.75%.
6. according to the preparation method described in claim 1, it is characterised in that the hydrolysis temperature of the solution described in step 2 is 50
℃。
7. according to the preparation method described in claim 1, it is characterised in that the enzymolysis time of the solution described in step 2 is
140min。
8. according to the preparation method described in claim 1, it is characterised in that the ultrasonic power of the solution described in step 3 is
360W。
9. according to the preparation method described in claim 1, it is characterised in that the ultrasonic time of the solution described in step 3 is
20min。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107158044A (en) * | 2017-05-09 | 2017-09-15 | 福建农林大学 | A kind of method for integrated extraction of ganoderma lucidum liquid extract |
| CN111973479A (en) * | 2020-07-16 | 2020-11-24 | 重庆珂蓝健康管理集团有限公司 | Massage eye cream |
| CN114717286A (en) * | 2022-04-29 | 2022-07-08 | 吉林农业大学 | Preparation method of mellea armillaria sporophore antioxidant peptide |
-
2016
- 2016-09-05 CN CN201610800639.7A patent/CN106220751A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107158044A (en) * | 2017-05-09 | 2017-09-15 | 福建农林大学 | A kind of method for integrated extraction of ganoderma lucidum liquid extract |
| CN111973479A (en) * | 2020-07-16 | 2020-11-24 | 重庆珂蓝健康管理集团有限公司 | Massage eye cream |
| CN114717286A (en) * | 2022-04-29 | 2022-07-08 | 吉林农业大学 | Preparation method of mellea armillaria sporophore antioxidant peptide |
| CN114717286B (en) * | 2022-04-29 | 2024-05-10 | 吉林农业大学 | Preparation method of hazel mushroom antioxidant peptide |
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Application publication date: 20161214 |