CN101613418A - A kind of method of extracting agaricus lentinan - Google Patents
A kind of method of extracting agaricus lentinan Download PDFInfo
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- CN101613418A CN101613418A CN200910151176A CN200910151176A CN101613418A CN 101613418 A CN101613418 A CN 101613418A CN 200910151176 A CN200910151176 A CN 200910151176A CN 200910151176 A CN200910151176 A CN 200910151176A CN 101613418 A CN101613418 A CN 101613418A
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Abstract
The invention discloses a kind of method of extracting agaricus lentinan, concrete steps are as follows: Twospore Mushroom → making beating → adjust pH → complex enzyme hydrolysis → blunt enzyme → microwave treatment → insulation → filtration → centrifugal → concentrating under reduced pressure → Sevage method Deproteinization → alcohol precipitation → centrifugal → membrane filtration → resin absorption → desorb → vacuum concentration → vacuum-drying → agaricus lentinan; The present invention has agaricus lentinan extraction yield height, the characteristics that dna purity is also high, and non-pollutant discharge in the technological process, energy consumption is low, and processing method is reasonable in design.
Description
Technical field
The present invention relates to a kind of method of extracting agaricus lentinan, microorganism belonging to genus chemistry macromolecular cpd isolation technique.
Background technology
Twospore Mushroom (Agaricus burnnescens peck), the formal name used at school mushroom, white mushroom or Mushroom belong to Mycophyta, Basidiomycotina, Hymenomycetes, Agaricales, Agaricus edibilis; Be a kind of wild edible fungus of eating the medicine dual-purpose, have the characteristics of high protein, lower fat, amino acid A wide selection of colours and designs, useful again spleen, clearing away the heart fire and tranquillizing, help food, control the effect of hemorrhoid, regulate human body metabolism in addition, strengthen the effect of human body constitution; Carbohydrate in the Twospore Mushroom is not only the important component and the energy substance of human body cell, but also has immunity, different physiological roles such as anticancer.
Chinese patent application numbers 200810222270.1 discloses " a kind of agaricus bisporus extract and preparation method thereof and application ", it be with Twospore Mushroom sporophore and water according to 1: after the volume ratio of 3-5 is mixed homogenate, in 75-95 ℃, the pH value is to extract 1-6 hour under the condition of 7-9, remove residue, obtain containing the Twospore Mushroom extracting solution of agaricus lentinan.The efficient of this method extraction agaricus lentinan is on the low side, and purity is not high.
Summary of the invention
Purpose of the present invention aims to provide a kind of method of extracting agaricus lentinan, makes agaricus lentinan extraction efficiency and purity all high, adopts the not only energy-conservation but also environmental protection of this method.
For achieving the above object, the present invention extracts the method for agaricus lentinan and is: Twospore Mushroom → making beating → accent pH value → complex enzyme hydrolysis → blunt enzyme → microwave treatment → insulation → filtration → centrifugal → concentrating under reduced pressure → Sevage method Deproteinization → alcohol precipitation → centrifugal → membrane filtration → resin absorption → desorb → vacuum concentration → vacuum-drying → agaricus lentinan; Concrete steps are as follows:
1. the Twospore Mushroom crushing and beating being become pasty state, is that 1: 10 ratio is made into dope in Twospore Mushroom than pure water, adds the lemon acid for adjusting pH value to 5.5-5.6 in dope;
2. add cellulase, hemicellulase and polygalacturonase in the dope that regulates the pH value, stirring action is 3 hours under 45 ℃ of constant temperatures, and dope is carried out complex enzyme hydrolysis, afterwards by 100 ℃ of blunt enzyme processing of moment;
3. the dope behind the complex enzyme hydrolysis is inserted in the microwave instrument, carry out the intermittence microwave treatment 3 times;
4. the liquid after the microwave treatment is placed 55 ℃ of water-bath insulations 30 minutes, use the filter cloth press filtration, again filter residue is washed after-filtration with hot distilled water, twice filtrate filtered merged;
5. filtrate was carried out centrifugal treating 10 minutes, supernatant liquor is placed be evaporated to 1/5 of original volume under the 600mmHg vacuum tightness;
6. use the Sevage method: use chloroform: add after the mixed of amylalcohol=4: 1 in the material behind the concentrating under reduced pressure and carry out jolting, making the protein denaturation in the material is insoluble state, removes protein with centrifuging then;
7. in removing proteinic material, add dehydrated alcohol, make dehydrated alcohol and mixing of materials form solution, transfer to whole concentration and contain 70% ethanol, under 4 ℃ of low temperature, left standstill 24 hours, pure precipitated liquid was carried out centrifugation 10 minutes;
8. the control material is in 32-35 ℃ of scope, and working pressure is 2.0MPa, selects for use molecular film to filter, and mould material is the ultra-filtration membrane of molecular weight cut-off 50,000, obtains rough agaricus lentinan;
9. will carry out pretreated styrene type resin and pack in the post, again with rough agaricus lentinan extracting solution upper prop near saturated carrying out, rate of adsorption 2BV/h; Carry out desorb with ethanol solution then, elution speed 1BV/h obtains stripping liquid; Stripping liquid is done near through the rotatory evaporator vacuum concentration, is put at last in 60 ℃ of vacuum drying ovens dry 3 hours, promptly obtains agaricus lentinan.
The present invention compared with prior art has agaricus lentinan extraction yield height, the characteristics that dna purity is also high, and non-pollutant discharge in the technological process, energy consumption is low, and processing method is reasonable in design.
Embodiment
Take by weighing 500 kilograms of Twospore Mushroomes, crushing and beating becomes pasty state, is made into dope with 10 times pure water, transfers pH value to 5.5-5.6 with citric acid again; Add cellulase 0.2 ten thousand U, hemicellulase 0.1 ten thousand U, polygalacturonase 0.4 ten thousand U by every gram Twospore Mushroom, stirring action is 3 hours under 45 ℃ of constant temperatures, carries out complex enzyme hydrolysis, afterwards by 100 ℃ of blunt enzyme processing of moment; Dope behind the complex enzyme hydrolysis is inserted in the microwave instrument, carry out the intermittence microwave treatment 3 times, each microwave treatment time is 2 seconds, and the intermittent time is 35 seconds, and microwave instrument power is 500W; Liquid after the microwave treatment is placed 55 ℃ of water-bath insulations 30 minutes, use the filter cloth press filtration, filter residue washs with hot distilled water, merging filtrate, filtrate was carried out centrifugation 10 minutes, the rotating speed of whizzer is 4800r/min, the supernatant liquor after the centrifugation is placed be evaporated to 1/5 of original volume under the 600mmHg vacuum tightness; Use the Sevage method, use chloroform: add in the material after the mixed of amylalcohol=4: 1 and carry out jolting, make the protein denaturation in the material become insoluble state, remove protein with centrifuging then; In removing proteinic material, add dehydrated alcohol, make dehydrated alcohol and mixing of materials form solution, transfer to whole concentration and contain 70% ethanol, under 4 ℃ of low temperature, left standstill 24 hours, pure precipitated liquid was carried out centrifugation 10 minutes, and the rotating speed of whizzer is 4800r/min; Control material again in 32-35 ℃ of scope, working pressure is 2.0MPa, selects for use molecular film to filter, and mould material is the ultra-filtration membrane of molecular weight cut-off 50,000, obtains rough agaricus lentinan; To carry out pretreated styrene type resin packs in the post, with extremely approaching saturated the carrying out of rough agaricus lentinan extracting solution upper prop, rate of adsorption 2BV/h, carry out desorb with ethanol solution then, elution speed 1BV/h obtains stripping liquid, and stripping liquid is done near through the rotatory evaporator vacuum concentration, be put at last in 60 ℃ of vacuum drying ovens dry 3 hours, and promptly obtained agaricus lentinan.
Claims (3)
1. method of extracting agaricus lentinan is characterized in that:
(1) the Twospore Mushroom crushing and beating being become pasty state, is that 1: 10 ratio is made into dope in Twospore Mushroom than pure water, adds the lemon acid for adjusting pH value to 5.5-5.6 in dope;
(2) add cellulase, hemicellulase and polygalacturonase in the dope that regulates the pH value, stirring action is 3 hours under 45 ℃ of constant temperatures, and dope is carried out complex enzyme hydrolysis, afterwards by 100 ℃ of blunt enzyme processing of moment;
(3) dope behind the complex enzyme hydrolysis is inserted in the microwave instrument, carry out the intermittence microwave treatment 3 times;
(4) liquid after the microwave treatment is placed 55 ℃ of water-bath insulations 30 minutes, use the filter cloth press filtration, again filter residue is washed after-filtration with hot distilled water, twice filtrate filtered merged;
(5) filtrate was carried out centrifugal treating 10 minutes, supernatant liquor is placed be evaporated to 1/5 of original volume under the 600mmHg vacuum tightness;
(6) use the Sevage method: use chloroform: add after the mixed of amylalcohol=4: 1 in the material behind the concentrating under reduced pressure and carry out jolting, making the protein denaturation in the material is insoluble state, removes protein with centrifuging then;
(7) in removing proteinic material, add dehydrated alcohol, make dehydrated alcohol and mixing of materials form solution, transfer to whole concentration and contain 70% ethanol, under 4 ℃ of low temperature, left standstill 24 hours, pure precipitated liquid was carried out centrifugation 10 minutes;
(8) the control material is in 32-35 ℃ of scope, and working pressure is 2.0MPa, selects for use molecular film to filter, and mould material is the ultra-filtration membrane of molecular weight cut-off 50,000, obtains rough agaricus lentinan;
(9) will carry out pretreated styrene type resin and pack in the post, again with rough agaricus lentinan extracting solution upper prop near saturated carrying out, rate of adsorption 2BV/h; Carry out desorb with ethanol solution then, elution speed 1BV/h obtains stripping liquid; Stripping liquid is done near through the rotatory evaporator vacuum concentration, is put at last in 60 ℃ of vacuum drying ovens dry 3 hours, promptly obtains agaricus lentinan.
2. the method for extraction agaricus lentinan according to claim 1 is characterized in that adding cellulase 0.2 ten thousand U, hemicellulase 0.1 ten thousand U, polygalacturonase 0.4 ten thousand U by every gram Twospore Mushroom.
3. the method for extraction agaricus lentinan according to claim 1 is characterized in that each microwave treatment time is 2 seconds, and the intermittent time is 35 seconds, and microwave instrument power is 500W.
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101836706A (en) * | 2010-04-29 | 2010-09-22 | 李洁 | Method for preparing mushroom noodles |
| CN101891836A (en) * | 2010-07-27 | 2010-11-24 | 乔德亮 | Method for extracting white mushroom lentinan assisted with ultrasonic wave |
| CN102038264A (en) * | 2010-12-31 | 2011-05-04 | 山西师范大学 | Preparation method for agaricus bisporus enzymatic hydrolysis peptide oral liquid |
| CN102070723A (en) * | 2010-12-03 | 2011-05-25 | 曹庸 | Agaricus bisporus polysaccharide with immunological activity as well as preparation method and application of agaricus bisporus polysaccharide |
| CN102391387A (en) * | 2011-11-07 | 2012-03-28 | 沈阳科思高科技有限公司 | Microwave chemical extraction method of active polysaccharide in higher plants or edible and medicinal fungi |
| CN102488278A (en) * | 2011-12-08 | 2012-06-13 | 北京东方兴企食品工业技术有限公司 | Composite fungus nutrient solution with bifunctional health care effect |
| CN102532331A (en) * | 2010-12-24 | 2012-07-04 | 苏州宝泽堂医药科技有限公司 | Method for extracting melastoma dodecandrum lour water-soluble polysaccharides under assistance of microwaves |
| CN102675479A (en) * | 2012-05-17 | 2012-09-19 | 华南理工大学 | Method for producing pleurotus nebrodensis polysaccharide by compound enzymatic hydrolysis technology |
| CN102838686A (en) * | 2012-09-28 | 2012-12-26 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN103549100A (en) * | 2013-11-05 | 2014-02-05 | 周瑞保 | Method for preparing five-mushroom crispy sugar |
| CN103864944A (en) * | 2013-11-06 | 2014-06-18 | 青海麟丰农业科技有限公司 | Method for preparing pholiota nameko polysaccharide crude products |
| CN104861079A (en) * | 2015-04-27 | 2015-08-26 | 浙江山狼谷旅游产业发展有限公司 | Microwave assisted lentinan extraction equipment and technology |
| CN104987428A (en) * | 2015-07-23 | 2015-10-21 | 中国科学院西北高原生物研究所 | Compound enzyme-microwave extraction method for optimizing Nitraria polysaccharide by response surface methodology |
| CN105315383A (en) * | 2015-10-19 | 2016-02-10 | 闽南师范大学 | Purified active polysaccharides extracted from agaricus bisporus can processing waste water and extracting and purifying technology and application thereof |
| CN105330759A (en) * | 2015-11-23 | 2016-02-17 | 福建裕兴果蔬食品开发有限公司 | Method for extracting mushroom polysaccharides |
| CN110613639A (en) * | 2019-10-22 | 2019-12-27 | 沈阳翔源科技有限公司 | Preparation method of agaricus bisporus extract |
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Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101836706A (en) * | 2010-04-29 | 2010-09-22 | 李洁 | Method for preparing mushroom noodles |
| CN101891836A (en) * | 2010-07-27 | 2010-11-24 | 乔德亮 | Method for extracting white mushroom lentinan assisted with ultrasonic wave |
| CN101891836B (en) * | 2010-07-27 | 2012-05-02 | 乔德亮 | Method for extracting white mushroom lentinan assisted with ultrasonic wave |
| CN102070723A (en) * | 2010-12-03 | 2011-05-25 | 曹庸 | Agaricus bisporus polysaccharide with immunological activity as well as preparation method and application of agaricus bisporus polysaccharide |
| CN102532331A (en) * | 2010-12-24 | 2012-07-04 | 苏州宝泽堂医药科技有限公司 | Method for extracting melastoma dodecandrum lour water-soluble polysaccharides under assistance of microwaves |
| CN102038264A (en) * | 2010-12-31 | 2011-05-04 | 山西师范大学 | Preparation method for agaricus bisporus enzymatic hydrolysis peptide oral liquid |
| CN102391387A (en) * | 2011-11-07 | 2012-03-28 | 沈阳科思高科技有限公司 | Microwave chemical extraction method of active polysaccharide in higher plants or edible and medicinal fungi |
| CN102488278A (en) * | 2011-12-08 | 2012-06-13 | 北京东方兴企食品工业技术有限公司 | Composite fungus nutrient solution with bifunctional health care effect |
| CN102675479A (en) * | 2012-05-17 | 2012-09-19 | 华南理工大学 | Method for producing pleurotus nebrodensis polysaccharide by compound enzymatic hydrolysis technology |
| CN102838686A (en) * | 2012-09-28 | 2012-12-26 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN102838686B (en) * | 2012-09-28 | 2014-11-12 | 天津商业大学 | Method for preparing high-purity agaricus blazei murrill polysaccharide |
| CN103549100A (en) * | 2013-11-05 | 2014-02-05 | 周瑞保 | Method for preparing five-mushroom crispy sugar |
| CN103864944A (en) * | 2013-11-06 | 2014-06-18 | 青海麟丰农业科技有限公司 | Method for preparing pholiota nameko polysaccharide crude products |
| CN103864944B (en) * | 2013-11-06 | 2016-01-13 | 青海麟丰农业科技有限公司 | A kind of preparation method of pholiota nameko polysaccharide crude product |
| CN104861079A (en) * | 2015-04-27 | 2015-08-26 | 浙江山狼谷旅游产业发展有限公司 | Microwave assisted lentinan extraction equipment and technology |
| CN104987428A (en) * | 2015-07-23 | 2015-10-21 | 中国科学院西北高原生物研究所 | Compound enzyme-microwave extraction method for optimizing Nitraria polysaccharide by response surface methodology |
| CN105315383A (en) * | 2015-10-19 | 2016-02-10 | 闽南师范大学 | Purified active polysaccharides extracted from agaricus bisporus can processing waste water and extracting and purifying technology and application thereof |
| CN105330759A (en) * | 2015-11-23 | 2016-02-17 | 福建裕兴果蔬食品开发有限公司 | Method for extracting mushroom polysaccharides |
| CN110613639A (en) * | 2019-10-22 | 2019-12-27 | 沈阳翔源科技有限公司 | Preparation method of agaricus bisporus extract |
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Application publication date: 20091230 |