CN108840958A - A method of extracting coriolan from rainbow conk - Google Patents
A method of extracting coriolan from rainbow conk Download PDFInfo
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- 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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Abstract
The present invention provides a kind of method that coriolan is extracted from rainbow conk, is related to biopharmaceutical technology, which includes degreasing, infiltration, enzymatic treatment, ultrasonic wave extraction, second extraction and alcohol precipitation.The extracting method is acted synergistically using degreasing, broken wall treatment, enzymatic treatment and ultrasound, microwave assisting method, has many advantages, such as that operating procedure is simple, saves solvent, product yield high, short processing time.
Description
Technical field
The present invention relates to biopharmaceutical technologies, and in particular to a kind of side that coriolan is extracted from rainbow conk
Method.
Background technique
Rainbow conk (Coriolusversicolor) is also known as Coriolus, thousand layers of mushroom, bright cloud leather lid bacterium etc., is subordinate to fungi
Door, Basidiomycotina, Polyporaceae, Coriolus Qu61 are a kind of widely distributed large-scale wood-rotting fungis, and rainbow conk is sweet in flavor, light, and property is micro-
It is cold, there is invigorating spleen to remove dampness, relieving cough and asthma, clearing heat and detoxicating, antitumor effect is the traditional rare medicinal herbs in China.Coriolan is
The principle active component of rainbow conk can activate and exempt to set cell, improve body's immunity, antitumor, reducing blood lipid and antibacterial, in addition,
Coriolan is significantly improved effect to learning and memory obstacle etc. caused by chronic stress, and therefore, coriolan can be made
For a kind of safely and effectively active material, it is applied in health food.
The extracting method of traditional coriolan mainly includes water extract method, acid or alkaline extraction etc..That is, hot water extraction method will
Rainbow conk is impregnated in the hot water, and high temperature decocts, and water-soluble polysaccharide is extracted from rainbow conk, but time-consuming for the method, the extraction of polysaccharide
Rate is lower, at high cost.Acid or alkaline extraction promote less the highest yield of coriolan, but cause seriously to pollute to environment.Cause
This, is badly in need of seeking a kind of extracting method of coriolan that can obtain high yield high-content.
In recent years, there are the new extractive techniques and method such as ultrasound or microblogging auxiliary extraction, super-pressure, enzyme process, still
The extraction effect of these methods is also very unsatisfactory.
Summary of the invention
Goal of the invention of the invention is, in view of the above-mentioned problems, a kind of method that coriolan is extracted from rainbow conk is provided, it should
Extracting method is acted synergistically using enzymatic treatment and ultrasonic wave added method, simple, product yield high, short processing time with operating procedure
The advantages that.
In order to achieve the above objectives, the technical scheme adopted by the invention is that：
A method of it extracting coriolan from rainbow conk, includes the following steps,
S1, degreasing：Manyzoned polypore sporophore is dried, coarse crushing；It is 80~90% that 5~10 times of bulking value scores, which are added,
Ethyl alcohol, refluxing extraction carries out degreasing under the conditions of 75~80 DEG C, obtains crude extract.
S2, infiltration：The crude extract is dried, broken wall, the water of 5~10 times of weight is then added, at 50~70 DEG C
1~3h of lower immersion, obtains pretreatment fluid.
S3, enzymatic treatment：The pH for adjusting the pretreatment fluid is 6~8, and the 3~8% of manyzoned polypore sporophore quality are then added
Complex enzyme, handles 1~3h at 40~55 DEG C, and then enzyme deactivation obtains extracting solution.
The complex enzyme includes that mass ratio is 2~5:1~3:3:1:2:1~3:1 cellulase, hemicellulase, fruit
Glue enzyme, zytase, ɑ-amylase, neutral proteinase and mannase.
S4, ultrasonic wave extraction：By 30~40min of the extracting solution ultrasonic wave extraction, Extracting temperature is 40~45 DEG C, then
Centrifugal filtration obtains supernatant I and residue.
S5, second extraction：Residue in step S4 is subjected to second extraction, 5~8 times of volumes are added to the residue
Water, 20~40min of microwave digestion at 60~80 DEG C are then centrifuged for filtering to obtain supernatant II.
S6, alcohol precipitation：Merge supernatant I and supernatant II obtain filtrate, by the filtrate decompression be concentrated into proper mass 40~
60%, 95% ethyl alcohol is added, makes ethyl alcohol final concentration of 75~85% in filtrate, stands 2~4h filtering, vacuum drying, crush, obtain
Coriolan.
Preferably, in step S1, manyzoned polypore sporophore coarse crushing is 40~50 mesh.
Preferably, in step S2, the crude extract is dried, broken wall is 400~600 mesh.
Preferably, in step S3, the cellulase, hemicellulase, pectase, zytase, ɑ-amylase, neutrality
The enzyme-activity unit of protease and mannase is 1~100,000 U/g.
Preferably, in step S3, the enzyme deactivation handles 10~20min at 85~95 DEG C.
Preferably, in step S4, ultrasonic power is 100~250W.
Preferably, in step S5, microwave frequency is 2400~2500MHz in the microwave digestion operation.
Preferably, in step S4 and S5, the centrifugal filtration process be at 6000~7000r/min centrifugation 10~
15min。
Due to the adoption of the above technical scheme, the invention has the advantages that：
1. it is of the invention from rainbow conk extract coriolan method, using degreasing, broken wall treatment, enzymatic treatment and ultrasound,
Microwave assisting method synergistic effect has many advantages, such as that operating procedure is simple, saves solvent, product yield high, short processing time.
Manyzoned polypore sporophore polysaccharide is extracted using conbined enzymolysis, effective component is on the one hand made more by degrading plant cell wall
It easily extracts to achieve the purpose that improve recovery rate or reduces solvent-oil ratio；On the other hand it can be directed to therein most of miscellaneous
Matter (starch, pectin, protein etc.) degradation selectivity, to be more conducive to the progress of later separation purifying.Complex enzyme uses cellulose
Enzyme, hemicellulase, pectase, zytase, ɑ-amylase, neutral proteinase and mannase are used in combination, Ke Yiti
High Degree of Enzymatic Hydrolysis shortens the time, improves extraction rate and yield.
2. the method for the invention for extracting coriolan from rainbow conk, using ultrasonic wave and microwave two-stage extracting mode, knot
Synthase processing and heating extraction, can greatly improve product yield.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment 1
A method of it extracting coriolan from rainbow conk, includes the following steps,
S1, degreasing：Manyzoned polypore sporophore is dried, coarse crushing；The ethyl alcohol of 8 times of weight, the volume point of the ethyl alcohol is added
Number is 85%, and refluxing extraction carries out degreasing under the conditions of 75 DEG C, obtains crude extract.Manyzoned polypore sporophore coarse crushing is 50 mesh.
S2, infiltration：The crude extract is dried, broken wall, broken wall is 500 mesh.Then the water of 6 times of weight is added,
2h is impregnated at 60 DEG C, obtains pretreatment fluid.
S3, enzymatic treatment：The pH for adjusting the pretreatment fluid is 7, and the compound of the 5% of manyzoned polypore sporophore quality is then added
Enzyme, handles 1~3h at 40~55 DEG C, and then enzyme deactivation obtains extracting solution.The enzyme deactivation is to handle 15min at 90 DEG C.
The complex enzyme includes that mass ratio is 3:2:3:1:2:2:1 cellulase, hemicellulase, pectase, wood are poly-
Carbohydrase, ɑ-amylase, neutral proteinase and mannase.The enzyme-activity unit of the cellulase is 60,000 U/g, hemicellulose
The enzyme-activity unit of enzyme is 30,000 U/g, and the enzyme-activity unit of pectase is 50,000 U/g, and the enzyme-activity unit of zytase is 80,000 U/g, ɑ-shallow lake
The enzyme-activity unit of powder enzyme is 100,000 U/g, and the enzyme-activity unit of neutral proteinase is 80,000 U/g and the enzyme-activity unit of mannase is 6
Ten thousand U/g.
S4, ultrasonic wave extraction：By the extracting solution ultrasonic wave extraction 35min, ultrasonic power 150W, Extracting temperature is
It 45 DEG C, is then centrifuged for filtering to obtain supernatant I and residue, centrifugal filtration process is to be centrifuged 12min at 6500r/min.
S5, second extraction：Residue in step S4 is subjected to second extraction, the water of 6 times of volumes is added to the residue,
Microwave digestion 30min at 70 DEG C is then centrifuged for filtering to obtain supernatant II.Microwave frequency is in the microwave digestion operation
2450MHz, the centrifugal filtration process are to be centrifuged 12min at 6500r/min.
S6, alcohol precipitation：Merge supernatant I and supernatant II and obtain filtrate, the filtrate decompression is concentrated into the 50% of proper mass,
95% ethyl alcohol is added, makes ethyl alcohol final concentration of 80% in filtrate, stands 3h filtering, vacuum drying, crush, obtain coriolan.
Embodiment 2
A method of it extracting coriolan from rainbow conk, includes the following steps,
S1, degreasing：Manyzoned polypore sporophore is dried, coarse crushing；The ethyl alcohol of 5 times of weight, the volume point of the ethyl alcohol is added
Number is 90%, and refluxing extraction carries out degreasing under the conditions of 80 DEG C, obtains crude extract.Manyzoned polypore sporophore coarse crushing is 40 mesh.
S2, infiltration：The crude extract is dried, broken wall, broken wall is 600 mesh.Then the water of 5 times of weight is added,
1h is impregnated at 70 DEG C, obtains pretreatment fluid.
S3, enzymatic treatment：The pH for adjusting the pretreatment fluid is 6, and the compound of the 3% of manyzoned polypore sporophore quality is then added
Enzyme handles 1h, then enzyme deactivation at 55 DEG C, obtains extracting solution.The enzyme deactivation is to handle 20min at 85 DEG C.
The complex enzyme includes that mass ratio is 5:3:3:1:2:1:1 cellulase, hemicellulase, pectase, wood are poly-
Carbohydrase, ɑ-amylase, neutral proteinase and mannase.The enzyme-activity unit of the cellulase is 80,000 U/g, hemicellulose
The enzyme-activity unit of enzyme is 60,000 U/g, and the enzyme-activity unit of pectase is 40,000 U/g, and the enzyme-activity unit of zytase is 50,000 U/g, ɑ-shallow lake
The enzyme-activity unit of powder enzyme is 60,000 U/g, and the enzyme-activity unit of neutral proteinase is 80,000 U/g and the enzyme-activity unit of mannase is 5
Ten thousand U/g.
S4, ultrasonic wave extraction：By the extracting solution ultrasonic wave extraction 40min, ultrasonic power 100W, Extracting temperature is
It 45 DEG C, is then centrifuged for filtering to obtain supernatant I and residue, centrifugal filtration process is to be centrifuged 10min at 7000r/min.
S5, second extraction：Residue in step S4 is subjected to second extraction, the water of 5 times of volumes is added to the residue,
Microwave digestion 20min at 80 DEG C is then centrifuged for filtering to obtain supernatant II.Microwave frequency is in the microwave digestion operation
2500MHz, the centrifugal filtration process are to be centrifuged 10min at 7000r/min.
S6, alcohol precipitation：Merge supernatant I and supernatant II and obtain filtrate, the filtrate decompression is concentrated into the 60% of proper mass,
95% ethyl alcohol is added, makes ethyl alcohol final concentration of 85% in filtrate, stands 2h filtering, vacuum drying, crush, obtain coriolan.
Embodiment 3
A method of it extracting coriolan from rainbow conk, includes the following steps,
S1, degreasing：Manyzoned polypore sporophore is dried, coarse crushing；The ethyl alcohol of 10 times of weight, the volume point of the ethyl alcohol is added
Number is 80%, and refluxing extraction carries out degreasing under the conditions of 75 DEG C, obtains crude extract.Manyzoned polypore sporophore coarse crushing is 50 mesh.
S2, infiltration：The crude extract is dried, broken wall, broken wall is 400 mesh.Then the water of 5 times of weight is added,
3h is impregnated at 50 DEG C, obtains pretreatment fluid.
S3, enzymatic treatment：The pH for adjusting the pretreatment fluid is 8, and the compound of the 8% of manyzoned polypore sporophore quality is then added
Enzyme handles 3h, then enzyme deactivation at 40 DEG C, obtains extracting solution.The enzyme deactivation is to handle 10min at 95 DEG C.
The complex enzyme includes that mass ratio is 2:3:3:1:2:3:1 cellulase, hemicellulase, pectase, wood are poly-
Carbohydrase, ɑ-amylase, neutral proteinase and mannase.The enzyme-activity unit of the cellulase is 100,000 U/g, hemicellulose
The enzyme-activity unit of enzyme is 60,000 U/g, and the enzyme-activity unit of pectase is 30,000 U/g, and the enzyme-activity unit of zytase is 40,000 U/g, ɑ-shallow lake
The enzyme-activity unit of powder enzyme is 30,000 U/g, and the enzyme-activity unit of neutral proteinase is 50,000 U/g and the enzyme-activity unit of mannase is 5
Ten thousand U/g.
S4, ultrasonic wave extraction：By the extracting solution ultrasonic wave extraction 30min, ultrasonic power 250W, Extracting temperature is
It 40 DEG C, is then centrifuged for filtering to obtain supernatant I and residue, centrifugal filtration process is to be centrifuged 15min at 6000r/min.
S5, second extraction：Residue in step S4 is subjected to second extraction, the water of 8 times of volumes is added to the residue,
Microwave digestion 40min at 60 DEG C is then centrifuged for filtering to obtain supernatant II.Microwave frequency is in the microwave digestion operation
2400MHz, the centrifugal filtration process are to be centrifuged 15min at 6000r/min.
S6, alcohol precipitation：Merge supernatant I and supernatant II and obtain filtrate, the filtrate decompression is concentrated into the 40% of proper mass,
95% ethyl alcohol is added, makes ethyl alcohol final concentration of 7% in filtrate, stands 4h filtering, vacuum drying, crush, obtain coriolan.
Comparative example 1
A method of extracting coriolan from rainbow conk, compared with Example 1, enzymatic treatment process uses single fibre
Tie up plain enzymatic treatment.
Comparative example 2
A method of coriolan is extracted from rainbow conk, compared with Example 1, in step S2, without broken wall treatment.
Comparative example 3
A method of extracting coriolan from rainbow conk does not have second extraction step compared with Example 1.
Comparative example 4
A method of coriolan is extracted from rainbow conk, compared with Example 1, without defatting step.
The method that embodiment 1-3 and comparative example 1-4 is respectively adopted extracts coriolan, and calculates yield, and result data is shown in
The following table 1.
1 embodiment 1-3 of table and comparative example 1-4 data comparison
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | |
| Polysaccharide yield/% | 21.45 | 20.35 | 20.56 | 10.25 | 12.56 | 11.52 | 13.68 |
From embodiment 1-3 and comparative example 1-4 can be seen that the present invention using degreasing, broken wall treatment, enzymatic treatment and ultrasound,
Microwave assisting method synergistic effect, polysaccharide yield are higher.Embodiment 1 and comparative example 1-4 can be seen that degreasing, broken wall treatment, at enzyme
Reason and ultrasonic wave added method synergistic effect, improve yield jointly, reduce either step, can all reduce yield.
Above description is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, it is all the present invention suggested by technical spirit under completed same changes or modifications change, should all belong to
In the covered the scope of the patents of the present invention.
Claims (8)
1. a kind of method for extracting coriolan from rainbow conk, which is characterized in that include the following steps,
S1, degreasing：Manyzoned polypore sporophore is dried, coarse crushing；The second that 5~10 times of bulking value scores are 80~90% is added
Alcohol, refluxing extraction carries out degreasing under the conditions of 75~80 DEG C, obtains crude extract；
S2, infiltration：The crude extract is dried, broken wall, the water of 5~10 times of weight is then added, is soaked at 50~70 DEG C
1~3h is steeped, pretreatment fluid is obtained；
S3, enzymatic treatment：The pH for adjusting the pretreatment fluid is 6~8, and the compound of the 3~8% of manyzoned polypore sporophore quality is then added
Enzyme, handles 1~3h at 40~55 DEG C, and then enzyme deactivation obtains extracting solution；
The complex enzyme includes that mass ratio is 2~5:1~3:3:1:2:1~3:1 cellulase, hemicellulase, pectase,
Zytase, ɑ-amylase, neutral proteinase and mannase；
S4, ultrasonic wave extraction：By 30~40min of the extracting solution ultrasonic wave extraction, Extracting temperature is 40~45 DEG C, is then centrifuged for
Filter to obtain supernatant I and residue；
S5, second extraction：Residue in step S4 is subjected to second extraction, the water of 5~8 times of volumes is added to the residue,
20~40min of microwave digestion at 60~80 DEG C is then centrifuged for filtering to obtain supernatant II；
S6, alcohol precipitation：Merge supernatant I and supernatant II and obtain filtrate, the filtrate decompression is concentrated into the 40~60% of proper mass,
95% ethyl alcohol is added, makes ethyl alcohol final concentration of 75~85% in filtrate, stands 2~4h filtering, vacuum drying, crush, obtain rainbow conk
Polysaccharide.
2. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that in step S1, rainbow conk
Fructification coarse crushing is 40~50 mesh.
3. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that in step S2, by institute
State crude extract be dried, broken wall be 400~600 mesh.
4. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that described in step S3
The enzyme-activity unit of cellulase, hemicellulase, pectase, zytase, ɑ-amylase, neutral proteinase and mannase
For 1~100,000 U/g.
5. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that described in step S3
Enzyme deactivation handles 10~20min at 85~95 DEG C.
6. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that in step S4, ultrasound
Wave power is 100~250W.
7. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that described in step S5
Microwave frequency is 2400~2500MHz in microwave digestion operation.
8. the method according to claim 1 for extracting coriolan from rainbow conk, which is characterized in that in step S4 and S5,
The centrifugal filtration process is that 10~15min is centrifuged at 6000~7000r/min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110776582A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Method for extracting β -glucan in sparassis crispa |
| CN110790849A (en) * | 2019-11-20 | 2020-02-14 | 湖北中烟工业有限责任公司 | Preparation method of Chinese yam mucopolysaccharide and application of Chinese yam mucopolysaccharide in cigarettes |
-
2018
- 2018-06-28 CN CN201810692039.2A patent/CN108840958A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110790849A (en) * | 2019-11-20 | 2020-02-14 | 湖北中烟工业有限责任公司 | Preparation method of Chinese yam mucopolysaccharide and application of Chinese yam mucopolysaccharide in cigarettes |
| CN110776582A (en) * | 2019-11-25 | 2020-02-11 | 福清市火麒麟食用菌技术开发有限公司 | Method for extracting β -glucan in sparassis crispa |
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Application publication date: 20181120 |