CN103570839A - Extraction method for Sparassia crispa polysaccharide - Google Patents
Extraction method for Sparassia crispa polysaccharide Download PDFInfo
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- CN103570839A CN103570839A CN201210248121.9A CN201210248121A CN103570839A CN 103570839 A CN103570839 A CN 103570839A CN 201210248121 A CN201210248121 A CN 201210248121A CN 103570839 A CN103570839 A CN 103570839A
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Abstract
The invention aims to provide a method for three-factor (ultrasonic wave, compound enzyme and alkaline process) combined extraction of polysaccharide from Sparassia crispa. The method effectively improves the extraction rate of polysaccharide and increases the added value of Sparassia crispa. The method comprises the steps of: selecting fresh and mildew-free Sparassia crispa fruit bodies as materials, and carrying out washing, drying and crushing, ultrasonic wall breaking, composite enzymatic hydrolysis, and alkaline extraction, thus eventually obtaining the polysaccharide.
Description
technical field
The present invention relates to take the method for the compound extraction polysaccharide of a kind of three factors (ultrasonic wave, prozyme and alkaline process) that the celestial mushroom of lotus is raw material.
Background technology
The celestial mushroom of lotus (Sparassia crispa (Wulf.) Fr.) has another name called Sparassis crispa, silk ball mushroom, Japan its petal mushroom of person.Celestial mushroom is similar to silk ball and gains the name lotus, its fragrance is pleasant, meat is tender and crisp, mouthfeel Jiamei, unique flavor, and food flavor is tasty, has the effects such as anticancer, immunomodulatory, raising hemopoietic function simultaneously, be a kind of macro fungi of dietotherapeutic of preciousness, in Japan, be described as the magical mushroom of illusion.
Antitumor main active ingredient 6-branch in the celestial mushroom polysaccharide of lotus body, 1,3-β-D dextran (SCG) be mushroom class, content can reach 50%, the content of its SCG is approximately three times of glossy ganoderma, four times of Brazilian mushroom.Studies show that, the antitumor action of the celestial mushroom of lotus is mainly realized by its activeconstituents SCG, and SCG is come antitumor by the main generation by stimulating cytokine.In the anticancer test of small white mouse, can 100% stop the development of cancer, and can significantly improve white blood cell count, activating immune system.
The celestial mushroom main active ingredient of lotus---polysaccharide body, does not exist only in cell, is buried in cell walls in a large number as cell walls support frame, discharges difficulty larger.The extracting method of edible fungi polysaccharide product mainly adopts single factor (water extraction, acid system, alkaline process etc.) extraction method at present, and all multi-methods are pure lower at polysaccharide yield, the problem that wastage of material is larger.
Summary of the invention
The object of the present invention is to provide a kind of method of take the compound extraction polysaccharide of three factors (ultrasonic wave, prozyme and alkaline process) that the celestial mushroom of lotus is raw material, the method has improved the extraction yield of polysaccharide effectively, has increased the added value of the celestial mushroom of lotus.
The present invention is the extracting method of the celestial mushroom polysaccharide of a kind of lotus, and selecting the fresh nothing celestial massee fruiting bodies of lotus that goes mouldy is material, through cleaning, dry pulverizing, supersonic wave wall breaking, complex enzyme hydrolysis, alkali, carries and finally obtains polysaccharide.The steps include:
1) cleaning, drying is pulverized: select fresh, unaged, without the celestial massee fruiting bodies of the lotus that goes mouldy, with clear water, clean; The celestial massee fruiting bodies of lotus is cut into the thin slice 50-55 ℃ that 0.2-0.4cm is thick dries to constant weight; 50-100 mesh sieves of pulverizing;
2) supersonic wave wall breaking: take the celestial mushroom powder of a certain amount of lotus, adding distil water, 15-60min is processed in solid-liquid ratio 1:10-70 under the ultrasound condition of power 100W-700W;
3) complex enzyme zymohydrolysis: it is 5.0-8.0 that the solution after supersound process is adjusted pH, temperature keeps 25-50 ℃, adds composite enzyme solution (proteolytic enzyme of the polygalacturonase of 0.5%-3.0%, the cellulase of 0.5%-3.0%, 0.5%-3.0%) to act on 1-4h;
4) alkali is carried: the full liquid of enzymolysis of previous step adds sodium hydroxide, make sodium hydroxide final concentration reach 1mol/L, under 50-70 ℃ of conditions, soak 1-3 hours, now solution is thick, centrifugal (4000-8000rpm) obtains supernatant liquor I, residue uses the 1mol/L sodium hydroxide solution of 2-5 times of volumes to carry once again again or repeatedly, centrifugal (4000-8000rpm) obtains supernatant liquor II; Merge supernatant liquor I and II, add acetic acid and adjust pH to 5.5-7, vacuum concentration to 1/5, add 3-5 times of volume dehydrated alcohol precipitation polysaccharide, hold over night in 4 ℃ of refrigerators, centrifugal (4000-8000rpm), obtain throw out, add appropriate distilled water and dissolve, be loaded in dialysis tubing, flowing water dialysis 1-3d, adds the dehydrated alcohol alcohol precipitation of 3-6 times, hold over night in 4 ℃ of refrigerators, centrifugal (4000-8000rpm), obtain polysaccharide precipitation, through vacuum lyophilization, obtain Crude polysaccharides.
Subordinate list 1 is the Data Comparison of extracting method of the present invention and ordinary method:
Subordinate list 1
Three factor methods are extracted and common method contrast at present
| Method | Crude polysaccharides weight (g) | Extraction yield (%) | Holosaccharide weight (g) | Polysaccharide content (%) |
| Common water extraction | 0.6928 | 2.70 | 0.3844 | 55.49 |
| High Temperature High Pressure water extraction | 2.7078 | 10.83 | 1.4400 | 53.18 |
| Cold buck is molten | 2.3605 | 9.20 | 0.4584 | 19.42 |
| Thermokalite is water-soluble | 3.4289 | 13.71 | 2.0017 | 58.38 |
| Three factor composite algorithms | 4.3085 | 17.23 | 2.6096 | 60.57 |
Feature of the present invention is to have adopted supersonic wave wall breaking, prozyme decomposition method and alkali to put forward a kind of complex method of several different methods combination, and the maximized productive rate that improves polysaccharide, has saved raw materials cost.
Embodiment
Embodiment 1:
1) cleaning, drying is pulverized: select fresh, unaged, without the celestial massee fruiting bodies of the lotus that goes mouldy, with clear water, clean; The celestial massee fruiting bodies of lotus is cut into 55 ℃ of thin slices that 0.2-0.4cm is thick dries to constant weight; 80 mesh sieves of pulverizing.
2) supersonic wave wall breaking: take the celestial mushroom powder of 25g lotus adding distil water, solid-liquid ratio 1:25 processes 20min under the ultrasound condition of power 500w.
3) complex enzyme zymohydrolysis: solution after supersound process is adjusted pH to 6.5, temperature keeps 45 ℃, adds composite enzyme solution (2% polygalacturonase, 2% cellulase, 2% proteolytic enzyme) effect 2h.
4) alkali is carried: the full liquid of enzymolysis of previous step adds sodium hydroxide, make sodium hydroxide final concentration reach 1mol/L, under 65 ℃ of conditions, soak 2 hours, now solution is thick, centrifugal (8000rpm) obtains supernatant liquor I, residue is carried once with the 1mol/L sodium hydroxide solution of 3 times of volumes more again, and centrifugal (8000rpm) obtains supernatant liquor II.Merge supernatant liquor I and II, add acetic acid and adjust pH to 6, vacuum concentration to 1/5, add 3 times of volume dehydrated alcohol precipitation polysaccharide, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain throw out, add appropriate distilled water and dissolve, be loaded in dialysis tubing, flowing water dialysis 2d, adds the dehydrated alcohol alcohol precipitation of 4 times, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain polysaccharide precipitation, through vacuum lyophilization, obtain Crude polysaccharides.
Embodiment 2:
1) cleaning, drying is pulverized: select fresh, unaged, without the celestial massee fruiting bodies of the lotus that goes mouldy, with clear water, clean; The celestial massee fruiting bodies of lotus is cut into 50 ℃ of thin slices that 0.2-0.4cm is thick dries to constant weight.60 mesh sieves of pulverizing.
2) supersonic wave wall breaking: take the celestial mushroom powder of 25g lotus adding distil water, solid-liquid ratio 1:15 processes 25min under the ultrasound condition of power 400W.
3) complex enzyme zymohydrolysis: solution after supersound process is adjusted pH to 6.5, temperature keeps 45 ℃, adds composite enzyme solution (1.5% polygalacturonase, 2.4% cellulase, 1.6% proteolytic enzyme) effect 3h.
4) alkali is carried: the full liquid of enzymolysis of previous step adds sodium hydroxide, make sodium hydroxide final concentration reach 1mol/L, under 55 ℃ of conditions, soak 2.5 hours, now solution is thick, centrifugal (8000rpm) obtains supernatant liquor I, residue carries twice again with the 1mol/L sodium hydroxide solution of 3 times of volumes again, and centrifugal (8000rpm) obtains supernatant liquor II.Merge supernatant liquor I and II, add acetic acid and adjust pH to 6, vacuum concentration to 1/5, add 3 times of volume dehydrated alcohol precipitation polysaccharide, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain throw out, add appropriate distilled water and dissolve, be loaded in dialysis tubing, flowing water dialysis 2d, adds the dehydrated alcohol alcohol precipitation of 4 times, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain polysaccharide precipitation, through vacuum lyophilization, obtain Crude polysaccharides.
Embodiment 3:
1) cleaning, drying is pulverized: select fresh, unaged, without the celestial massee fruiting bodies of the lotus that goes mouldy, with clear water, clean; The celestial massee fruiting bodies of lotus is cut into 55 ℃ of thin slices that 0.2-0.4cm is thick dries to constant weight.80 mesh sieves of pulverizing.
2) supersonic wave wall breaking: take the celestial mushroom powder of 25g lotus adding distil water, solid-liquid ratio 1:40 processes 20min under the ultrasound condition of power 500w.
3) complex enzyme zymohydrolysis: solution after supersound process is adjusted pH to 6.5, temperature keeps 45 ℃, adds composite enzyme solution (2.5% polygalacturonase, 2.5% cellulase, 2.0% proteolytic enzyme) effect 1.5h.
4) alkali is carried: the full liquid of enzymolysis of previous step adds sodium hydroxide, make sodium hydroxide final concentration reach 1mol/L, under 65 ℃ of conditions, soak 2 hours, now solution is thick, centrifugal (8000rpm) obtains supernatant liquor I, residue is carried once with the 1mol/L sodium hydroxide solution of 3 times of volumes more again, and centrifugal (8000rpm) obtains supernatant liquor II.Merge supernatant liquor I and II, add acetic acid and adjust pH to 6, vacuum concentration to 1/5, add 3 times of volume dehydrated alcohol precipitation polysaccharide, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain throw out, add appropriate distilled water and dissolve, be loaded in dialysis tubing, flowing water dialysis 2d, adds the dehydrated alcohol alcohol precipitation of 4 times, hold over night in 4 ℃ of refrigerators, centrifugal (8000rpm), obtain polysaccharide precipitation, through vacuum lyophilization, obtain Crude polysaccharides.
Obviously, above-described embodiment is only for clearly demonstrating example of the present invention, embodiment 1 is preferably embodiment of the present invention, but embodiments of the present invention are not restricted to the described embodiments, to those of ordinary skill in the art, can make on the basis of the above description other multi-form variation and change.Cannot give all embodiments exhaustively here, everyly belong to the row that apparent variation that technical solution of the present invention amplifies out and change are included in protection scope of the present invention.
Subordinate list 2 is data comparisons of three embodiment of the present invention:
Subordinate list 2
Three factor methods are extracted three embodiment comparisons of the celestial mushroom polysaccharide of lotus
| Method | Crude polysaccharides weight (g) | Extraction yield (%) | Holosaccharide weight (g) | Polysaccharide content (%) |
| Embodiment 1 | 4.3085 | 17.23 | 2.6096 | 60.57 |
| Embodiment 2 | 3.7078 | 14.83 | 2.1301 | 57.45 |
| Embodiment 3 | 4.0605 | 16.24 | 2.3822 | 58.67 |
Claims (1)
1. an extracting method for the celestial mushroom polysaccharide of lotus, the steps include:
Cleaning, drying is pulverized: select fresh, unaged, without the celestial massee fruiting bodies of the lotus that goes mouldy, with clear water, clean; The celestial massee fruiting bodies of lotus is cut into the thin slice 50-55 ℃ that 0.2-0.4cm is thick dries to constant weight; 50-100 mesh sieves of pulverizing;
Supersonic wave wall breaking: take the celestial mushroom powder of a certain amount of lotus, adding distil water, 15-60min is processed in solid-liquid ratio 1:10-70 under the ultrasound condition of power 100W-700W;
Complex enzyme zymohydrolysis: it is 5.0-8.0 that the solution after supersound process is adjusted pH, temperature keeps 25-50 ℃, adds composite enzyme solution (proteolytic enzyme of the polygalacturonase of 0.5%-3.0%, the cellulase of 0.5%-3.0%, 0.5%-3.0%) to act on 1-4h;
Alkali is carried: the full liquid of enzymolysis of previous step adds sodium hydroxide, make sodium hydroxide final concentration reach 1mol/L, under 50-70 ℃ of conditions, soak 1-3 hours, now solution is thick, centrifugal (4000-8000rpm) obtains supernatant liquor I, residue uses the 1mol/L sodium hydroxide solution of 2-5 times of volumes to carry once again again or repeatedly, centrifugal (4000-8000rpm) obtains supernatant liquor II; Merge supernatant liquor I and II, add acetic acid and adjust pH to 5.5-7, vacuum concentration to 1/5, add 3-5 times of volume dehydrated alcohol precipitation polysaccharide, hold over night in 4 ℃ of refrigerators, centrifugal (4000-8000rpm), obtain throw out, add appropriate distilled water and dissolve, be loaded in dialysis tubing, flowing water dialysis 1-3d, adds the dehydrated alcohol alcohol precipitation of 3-6 times, hold over night in 4 ℃ of refrigerators, centrifugal (4000-8000rpm), obtain polysaccharide precipitation, through vacuum lyophilization, obtain Crude polysaccharides.
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| CN201210248121.9A CN103570839A (en) | 2012-07-18 | 2012-07-18 | Extraction method for Sparassia crispa polysaccharide |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105542026A (en) * | 2016-01-19 | 2016-05-04 | 济南大学 | Method for efficiently extracting okra polysaccharide |
| CN106749744A (en) * | 2017-02-13 | 2017-05-31 | 重庆中烟工业有限责任公司 | A kind of tobacco leaf polyoses extract and its extracting method and application |
| CN109957039A (en) * | 2019-03-22 | 2019-07-02 | 辽宁大学 | A kind of Sparassia crispa polysaccharide Extraction technique optimization method and extracting method |
| CN110835380A (en) * | 2019-12-12 | 2020-02-25 | 福建万菇生物科技有限公司 | Process for extracting sparassis crispa polysaccharide by microwave-assisted method |
| CN110835379A (en) * | 2019-12-12 | 2020-02-25 | 福建万菇生物科技有限公司 | Process for extracting sparassis crispa polysaccharide by ultrahigh pressure auxiliary method |
| CN110981987A (en) * | 2019-12-12 | 2020-04-10 | 福建万菇生物科技有限公司 | Technology for extracting sparassis crispa polysaccharide by ultrasonic-assisted method |
| CN114478819A (en) * | 2022-02-25 | 2022-05-13 | 辽宁大学 | Sparassia crispa polysaccharide and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002262820A (en) * | 2001-03-07 | 2002-09-17 | Isao Horiuchi | Method for extracting active ingredient of mushrooms |
| CN101112396A (en) * | 2006-07-24 | 2008-01-30 | 中岛三博 | Extract of sparassis crispa MH-3 bacterial strain and method for preparing the same and use thereof |
| CN101766656A (en) * | 2008-12-31 | 2010-07-07 | 陈秀男 | Combination containing mushroom polysaccharide and preparation method thereof |
-
2012
- 2012-07-18 CN CN201210248121.9A patent/CN103570839A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002262820A (en) * | 2001-03-07 | 2002-09-17 | Isao Horiuchi | Method for extracting active ingredient of mushrooms |
| CN101112396A (en) * | 2006-07-24 | 2008-01-30 | 中岛三博 | Extract of sparassis crispa MH-3 bacterial strain and method for preparing the same and use thereof |
| CN101766656A (en) * | 2008-12-31 | 2010-07-07 | 陈秀男 | Combination containing mushroom polysaccharide and preparation method thereof |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105542026A (en) * | 2016-01-19 | 2016-05-04 | 济南大学 | Method for efficiently extracting okra polysaccharide |
| CN106749744A (en) * | 2017-02-13 | 2017-05-31 | 重庆中烟工业有限责任公司 | A kind of tobacco leaf polyoses extract and its extracting method and application |
| CN106749744B (en) * | 2017-02-13 | 2019-10-11 | 重庆中烟工业有限责任公司 | A kind of tobacco leaf polyoses extract and its extracting method and application |
| CN109957039A (en) * | 2019-03-22 | 2019-07-02 | 辽宁大学 | A kind of Sparassia crispa polysaccharide Extraction technique optimization method and extracting method |
| CN110835380A (en) * | 2019-12-12 | 2020-02-25 | 福建万菇生物科技有限公司 | Process for extracting sparassis crispa polysaccharide by microwave-assisted method |
| CN110835379A (en) * | 2019-12-12 | 2020-02-25 | 福建万菇生物科技有限公司 | Process for extracting sparassis crispa polysaccharide by ultrahigh pressure auxiliary method |
| CN110981987A (en) * | 2019-12-12 | 2020-04-10 | 福建万菇生物科技有限公司 | Technology for extracting sparassis crispa polysaccharide by ultrasonic-assisted method |
| CN110981987B (en) * | 2019-12-12 | 2021-08-20 | 福建万菇生物科技有限公司 | Technology for extracting sparassis crispa polysaccharide by ultrasonic-assisted method |
| CN114478819A (en) * | 2022-02-25 | 2022-05-13 | 辽宁大学 | Sparassia crispa polysaccharide and application thereof |
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Application publication date: 20140212 |