CN101560264A - Process combination for extracting polysaccharide in golden mushroom mycelium cells and determination method thereof - Google Patents
Process combination for extracting polysaccharide in golden mushroom mycelium cells and determination method thereof Download PDFInfo
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Abstract
The invention relates to a process combination for extracting polysaccharide in golden mushroom mycelium cells. The solid-liquid ratio according to weight is 1: 10-1: 60, the adding amount of complex enzyme accounts for 0.5 percent to 5.0 percent of the weight of the raw materials, the range of ultrasonic power is 100w to 500w, the range of enzymolysis time is 40min to 100min, the range of enzymolysis temperature is 30 DEG C to 60 DEG C, and the range of enzymolysis pH value is 4.0 to 8.0. The determination method of the process combination for extracting polysaccharide in golden mushroom mycelium cells is as follows: high-quality mycelium is selected to be deeply fermented, thus obtaining mycelium dry powder; the solid-liquid ratio is determined; the adding amount of complex enzyme is determined; and the optimal extracting process combination for extracting mycelium dry powder polysaccharide in an ultrasonic-complex enzyme method is determined.
Description
Technical field
The present invention relates to the bio-pharmaceuticals production technology, specifically a kind of process combination and definite method thereof of extracting polysaccharide in golden mushroom mycelium cells.
Background technology
Needle mushroom formal name used at school hair handle money bacterium, needle mushroom are the edible mushroomss of autumn and winter and early spring cultivation, with its cap sliding tender, handle is crisp, nutritious, delicious agreeable to the taste and be world-famous for.According to surveying and determination, the amino acid whose content of needle mushroom is very abundant, and the content that is higher than general mushroom class, especially Methionin is high especially, and Methionin has the function that promotes children ' s intelligence development.Contain protein 8.87% in the needle mushroom dry product, carbohydrate 60.2%, robust fibre reaches 7.4%, often eats and can prevent and treat Peptic Ulcers.Research recently shows that again contained a kind of material has good antitumous effect in the needle mushroom.Needle mushroom is a kind of acat, is again protective foods preferably.
Flammulina velutipes is widely used in fields such as medicine, food, market demand grows with each passing day, but the production of flammulina velutipes nearly all is to extract from sporophore at present, the needle mushroom wild resource lacks, the tame cycle is long, floor space is big, is subject to seasonal restrictions again, thereby has influenced making full use of of needle mushroom.Utilize the submerged fermentation technology to produce the golden mushroom mycelium polysaccharide, its biological effect and needle mushroom fruitbody polysaccharide much at one, and it is with short production cycle, cost is low, output is big, has industrialized prospect of production.
The golden mushroom mycelium polysaccharide that forms in submerged fermentation divides exocellular polysaccharide and intracellular polyse two portions, and intracellular polyse has accounted for 60-90%, is the main source of flammulina velutipes mycelium polysaccharides.Extraction polysaccharide mode commonly used has hot water extraction, acid leaching extraction method, alkali extraction method, enzyme extraction method etc.Because the existence of mycelial cell wall, intracellular component is difficult to be discharged into smoothly outside the born of the same parents, non-polysaccharide materials such as pectin substance that cell is inside and outside and protein also make extracting efficiency of polysaccharides and purity be affected, traditional some extract that polysaccharide modes cause leaching process length consuming time, the extraction yield of flammulina velutipes low, polysaccharide content is relatively low in the extract, have perhaps destroyed the molecular activity structure picture of polysaccharide because of chemical reagents reaction.Therefore, seeking efficiently, extraction method of polysaccharides is that development golden mushroom mycelium polysaccharide is the key in this field.
Natural medicinal ingredients mostly is product in the cell greatly, how need carry out cytoclasis during extraction, and existing mechanical and chemical process is difficult to obtain the ideal crushing effect sometimes.Ultrasonic technology is applied to the extraction of golden mushroom mycelium polysaccharide, be effects such as the judder that utilizes ultrasonic wave to produce, high acceleration, intensive cavitation effect, stirring, quicken effective constituent and enter solvent, thereby raising leaching yield, shorten extraction time, can avoid simultaneously proposing the influence of composition.This technology is widely used in Chinese medicine extracts active ingredients, separation and preparation technology.
In the leaching process of mycelium polysaccharides, also can utilize suitable enzyme, under relatively mild condition, cell tissue is decomposed, quicken the release of effective constituent, thereby improve polysaccharide yield.The prozyme of cellulase, polygalacturonase and proteolytic enzyme composition is used for the extraction of polysaccharide, to remove the compositions such as pectin, Mierocrystalline cellulose and protein on cell walls born of the same parents and the cell membrane, make mycelial cell break smoothly, help promoting the stripping of intracellular polyse, improve extraction yield, shorten extraction time, reduce energy consumption and effectively preserve the bioactive purpose of polysaccharide to reach.But can improve production cost owing to add enzyme extraction, bring difficulty also for simultaneously the separation of product, on addition, want strict control.
Summary of the invention
The objective of the invention is to, utilize ultrasonic wave the golden mushroom mycelium tissue to be handled as supplementary means, the prozyme of forming with cellulase, polygalacturonase and proteolytic enzyme obtains flammulina velutipes to golden mushroom mycelium infusion enzymolysis simultaneously, a kind of extraction effect better than ordinary method that have is provided, extracts polysaccharide in golden mushroom mycelium cells technology and definite method thereof with lower cost.
Technical scheme of the present invention is: a kind of process combination of extracting polysaccharide in golden mushroom mycelium cells, solid-liquid ratio is 1 by mass ratio: 10-1: 60, the prozyme add-on is between the 0.5%-5.0% of raw materials quality, the ultrasonic power value range is between the 100W-500W, the enzymolysis time value range is between the 40min-100min, the hydrolysis temperature value range is between 30 ℃-60 ℃, and enzymolysis pH value value range is between the 4.0-8.0.
Described a kind of definite method of extracting the process combination of polysaccharide in golden mushroom mycelium cells is:
1) the mycelium dry powder of selected fine golden mushroom mycelium submerged fermentation gained
2) determine solid-liquid ratio
The mycelium dry powder that step 1) is selected fully dissolves in the water, and the reasonable solid-liquid ratio of golden mushroom mycelium was at 1: 10 (g: g)-1: 60 (g: g).Under this solid-liquid ratio condition, carry out hot water lixiviate polysaccharide experiment, the control extraction temperature is in 50～95 ℃, lixiviate pH value 4.0～7.5, extraction time 40min～120min scope.After 5～15 times of the vat liquor evaporation concentration, add 95% ethanol sedimentation of 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, and vacuum-drying is to constant weight, weighing.By relatively, determine the highest solid-liquid ratio of polysaccharide in golden mushroom mycelium cells extraction yield that the hot water lixiviate obtains, be exactly the best solid-liquid ratio of this kind.
3) determine the addition of prozyme
According to top step 2) in the vat liquor raw material that is made into of best solid-liquid ratio in, add rational prozyme amount, scope is between the 0.5%-5.0% of vat liquor raw materials quality.Under suitable enzymatic hydrolysis condition, carry out experiment of single factor, determine the addition of the suitableeest prozyme.Under the addition condition of this prozyme, the control extraction temperature is in 40～65 ℃, lixiviate pH value 4.0～6.5, extraction time 40min～80min scope.After 5～15 times of the vat liquor evaporation concentration, add 95% ethanol sedimentation of 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, and vacuum-drying is to constant weight, weighing.By relatively, determine the addition of the prozyme that the polysaccharide in golden mushroom mycelium cells extraction yield is the highest, both be the addition of the best complex enzyme of this kind.
4) determine that ultrasonic wave-prozyme solution extracts the optimum extraction process combination of this mycelium dry powder polysaccharide
The complex enzyme hydrolysis that carries out needle mushroom fermentation mycelium polysaccharide under the ultrasonic wave booster action extracts.In step 2) and 3) solid-liquid ratio and the condition of prozyme addition under, with ultrasonic power (factor A), enzymolysis time (factor B), hydrolysis temperature (factor C), enzymolysis pH value (factor C) is analytic target, carry out the experiment of 4 factors, 3 horizontal quadratures, obtain the optimum extraction process combination.According to the previous experiments result, the scope of experiment of factor A
Value is between the 100W-500W, and the scope of experiment value of factor B is between the 40min-100min, and the scope of experiment value of factor C is between 30 ℃-60 ℃, and the scope of experiment value of factor D is between the 4.0-8.0.Extract under combination process condition at this, carry out mycelium polysaccharides and extract, after 5～15 times of the vat liquor evaporation concentration, add 95% ethanol sedimentation of 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, and vacuum-drying is to constant weight, weighing.By relatively, determine the highest process combination of polysaccharide in golden mushroom mycelium cells extraction yield, both be the optimum extraction process combination of this mycelium dry powder.
The present invention compared with prior art has the following advantages:
1, judder, the speed of superelevation, the intensive cavitation effect that utilizes ultrasonic wave to produce can quicken effective ingredient and enter solvent.
2, these non-polysaccharide fractions have been decomposed in the adding of prozyme effectively, under the synergy of ultrasonic wave mechanism, can improve the yield of polysaccharide more up hill and dale, shorten extraction time, save solvent, help protecting polysaccharide effective constituent.
3, technology can significantly reduce production costs in the extraction process of golden mushroom mycelium polysaccharide is transformed, and improves separation efficiency, and is adapted at applying in the suitability for industrialized production.
Embodiment
The golden mushroom mycelium dry powder polysaccharide extracting process combination that Qingyuan County, Zhejiang Province grid medicinal fungus company limited produces, its composition is: solid-liquid ratio is 1: 30 by mass ratio, the prozyme add-on is 2%, ultrasonic power is 200W, enzymolysis time 50min, 55 ℃ of hydrolysis temperatures, enzymolysis pH value 5.0.
Described a kind of definite method of extracting the polysaccharide in golden mushroom mycelium cells process combination is:
1) the golden mushroom mycelium dry powder of selected Zhejiang Province Qingyuan County grid medicinal fungus company production is as extracting raw material.
2) determine solid-liquid ratio
Extract polysaccharide by the hot water extraction.Choosing the mycelium dry powder 100g of step 1), is analytical factor to add the water multiple, and 10,20,30,40,50,60 times (quality multiples) are set, and is heated to 90 ℃ and keeps 3 hours, filters.Filtrate is concentrated into 100ml at rotatory evaporator, is cooled to normal temperature.95% ethanol that adds 400ml stirred 10 minutes with glass stick in concentrated solution simultaneously, with centrifugal 2 minutes of mixed solution 1000rmp, removed supernatant liquor, the mycelium polysaccharides precipitation, vacuum-drying weighing to the constant weight.After finding to add the water of 30 times of weight (3000g), the multiple extraction acquisition polysaccharide quantitative changeization that increases water again is little, so determine that solid-liquid ratio is 1: 30.
3) determine the addition of prozyme
Extract polysaccharide by the prozyme solution.With the compound enzymic preparation addition is empirical factor, at solid-liquid ratio is 1: 30,45 ℃ of temperature, pH6.5, under the condition of enzymolysis time 80min, the prozyme addition is set to 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5% (mass percent of mycelium dry powder relatively), behind the enzymolysis time 80min, and 95 ℃ of heating enzyme 5min that goes out, 80 ℃ of hot water continue lixiviate 60min, filter the rear filtrate treatment step with 1, the polysaccharide in golden mushroom mycelium cells of acquisition, weighing.Find that the prozyme addition is set to after 2.0%, increase the prozyme addition again, the golden mushroom mycelium intracellular polyse that the enzymolysis lixiviate obtains changes little, so determine that the prozyme addition is 2.0%.
4) optimum extraction process of determining this mycelium dry powder makes up
Extract polysaccharide by ultrasonic wave-prozyme solution.At solid-liquid ratio is 1: 30, under the condition of enzyme concentration 2.0%, with ultrasonic power (factor A), enzymolysis time (factor B), hydrolysis temperature (factor C), enzymolysis pH value (factor C) is analytic target, carries out the experiment of 4 factors, 3 horizontal quadratures, and quadrature level of factor table is as follows:
After the operation of the ultrasonic wave under each Orthogonal Composite condition-prozyme solution, 95 ℃ of heating of enzymolysis time enzyme 5min that goes out, 80 ℃ of hot water continue lixiviate 60min, filter the rear filtrate treatment step with 1, the golden mushroom mycelium intracellular polyse of acquisition, weighing.
Test-results shows, at A
2B
1C
3D
1The process combination condition under, polysaccharide extract rate is the highest in the mycelium cell, promptly when solid-liquid ratio be 1: 30, under enzyme concentration 2.0% condition, ultrasonic power is 200W, enzymolysis time 50min, 55 ℃ of hydrolysis temperatures, during enzymolysis pH value 5.0, polysaccharide extract rate is the highest in the mycelium cell.
Claims (2)
1, a kind of process combination of extracting polysaccharide in golden mushroom mycelium cells, it is characterized in that each component is: solid-liquid ratio is 1 by mass ratio: 10-1: 60, the prozyme add-on is between the 0.5%-5.0% of raw materials quality, the ultrasonic power value range is between the 100W-500W, the enzymolysis time value range is between the 40min-100min, the hydrolysis temperature value range is between 30 ℃-60 ℃, and enzymolysis pH value value range is between the 4.0-8.0.
2, according to the described a kind of definite method of extracting the process combination of polysaccharide in golden mushroom mycelium cells of claim 1, it is characterized in that:
1) the mycelium dry powder of selected fine mycelium submerged fermentation gained
2) determine solid-liquid ratio
The mycelium dry powder that step 1) is selected fully dissolves in the water, the reasonable solid-liquid ratio of golden mushroom mycelium was at 1: 10 (g: g)-1: 60 (g: g), under this solid-liquid ratio condition, carry out the experiment of hot water lixiviate polysaccharide, the control extraction temperature is 50～95 ℃, lixiviate pH value 4.0～7.5, in extraction time 40min～120min scope, after 5～15 times of the vat liquor evaporation concentration, 95% ethanol sedimentation that adds 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, vacuum-drying is to constant weight, and weighing is by comparing, determining the highest solid-liquid ratio of polysaccharide in golden mushroom mycelium cells extraction yield that the hot water lixiviate obtains, is exactly the best solid-liquid ratio of this kind;
3) determine the addition of prozyme
According to top step 2) in the vat liquor raw material that is made into of best solid-liquid ratio in, add rational prozyme amount, scope is between the 0.5%-5.0% of vat liquor raw materials quality, under suitable enzymatic hydrolysis condition, carry out experiment of single factor, determine the addition of the suitableeest prozyme, under the addition condition of this prozyme, the control extraction temperature is in 40～65 ℃, lixiviate pH value 4.0～6.5, extraction time 40min～80min scope.After 5～15 times of the vat liquor evaporation concentration, 95% ethanol sedimentation that adds 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, vacuum-drying is to constant weight, weighing, by relatively, determine the addition of the prozyme that the polysaccharide in golden mushroom mycelium cells extraction yield is the highest, both be the addition of the best complex enzyme of this kind;
4) determine that ultrasonic wave-prozyme solution extracts the optimum extraction process of this mycelium dry powder polysaccharide and be combined in the complex enzyme hydrolysis that carries out needle mushroom fermentation mycelium polysaccharide under the ultrasonic wave booster action and extract, in step 2) and 3) solid-liquid ratio and the condition of prozyme addition under, with ultrasonic power (factor A), enzymolysis time (factor B), hydrolysis temperature (factor C), enzymolysis pH value (factor C) is an analytic target, carry out the experiment of 4 factors, 3 horizontal quadratures, the combination of acquisition optimum extraction process, according to the previous experiments result, the scope of experiment value of factor A is between the 100W-500W, the scope of experiment value of factor B is between the 40min-100min, the scope of experiment value of factor C is between 30 ℃-60 ℃, the scope of experiment value of factor D is between the 4.0-8.0, extract under the combination process condition at this, carrying out mycelium polysaccharides extracts, after 5～15 times of the vat liquor evaporation concentration, 95% ethanol sedimentation that adds 4 times of volumes, 2000rmp is centrifugal, and supernatant liquor is removed in the back, vacuum-drying is to constant weight, weighing, by comparing, determining the highest process combination of polysaccharide in golden mushroom mycelium cells extraction yield, both has been the optimum extraction process combination of this mycelium dry powder.
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| CN102174615A (en) * | 2011-03-16 | 2011-09-07 | 浙江省农业科学院 | Method for producing fungal polysaccharide with residues after extraction of polysaccharide using needle mushroom |
| CN102702378A (en) * | 2012-06-29 | 2012-10-03 | 南京财经大学 | Method for extracting polysaccharides with cell immunocompetence from needle mushroom root waste materials |
| CN103073648A (en) * | 2012-09-11 | 2013-05-01 | 华南理工大学 | Method for increasing antioxidation activity of flammulina velutipes polysaccharide |
| CN103113487A (en) * | 2013-02-21 | 2013-05-22 | 嘉兴职业技术学院 | Method for preparing flammulina velutipes polysaccharides and protein efficiently synchronously |
| CN103788229A (en) * | 2014-02-19 | 2014-05-14 | 浙江省农业科学院 | Method for simultaneously extracting polysaccharide and polyphenol from needle mushrooms |
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| CN104387485A (en) * | 2014-11-12 | 2015-03-04 | 绥化学院 | Method for extracting polysaccharides in flammulina velutipes by synergism of complex enzymes and high-pressure hot water extraction process |
| CN107624513A (en) * | 2017-10-25 | 2018-01-26 | 山东惠民齐发果蔬有限责任公司 | It is a kind of rich in the edible and medical fungi cultural hypha method of polysaccharide and application |
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| CN115381087A (en) * | 2022-06-22 | 2022-11-25 | 深圳市维龄可伴生物科技有限公司 | Efficient extraction process and application of needle mushroom powder |
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| CN102174615B (en) * | 2011-03-16 | 2013-06-26 | 浙江省农业科学院 | Method for producing fungal polysaccharide with residues after extraction of polysaccharide using needle mushroom |
| CN102174615A (en) * | 2011-03-16 | 2011-09-07 | 浙江省农业科学院 | Method for producing fungal polysaccharide with residues after extraction of polysaccharide using needle mushroom |
| CN102702378B (en) * | 2012-06-29 | 2014-11-05 | 南京财经大学 | Method for extracting polysaccharides with cell immunocompetence from needle mushroom root waste materials |
| CN102702378A (en) * | 2012-06-29 | 2012-10-03 | 南京财经大学 | Method for extracting polysaccharides with cell immunocompetence from needle mushroom root waste materials |
| CN103073648A (en) * | 2012-09-11 | 2013-05-01 | 华南理工大学 | Method for increasing antioxidation activity of flammulina velutipes polysaccharide |
| CN103073648B (en) * | 2012-09-11 | 2016-01-20 | 华南理工大学 | A kind of method improving flammulina velutipes anti-oxidant activity |
| CN103113487A (en) * | 2013-02-21 | 2013-05-22 | 嘉兴职业技术学院 | Method for preparing flammulina velutipes polysaccharides and protein efficiently synchronously |
| CN103788223B (en) * | 2014-01-16 | 2016-05-04 | 山东省农业科学院农产品研究所 | A kind of extracting method of flammulina velutipes |
| CN103788223A (en) * | 2014-01-16 | 2014-05-14 | 山东省农业科学院农产品研究所 | Extraction method for flammukinan |
| CN103788229B (en) * | 2014-02-19 | 2016-02-17 | 浙江省农业科学院 | A kind of method simultaneously extracting polysaccharide and polyphenol from needle mushroom |
| CN103788229A (en) * | 2014-02-19 | 2014-05-14 | 浙江省农业科学院 | Method for simultaneously extracting polysaccharide and polyphenol from needle mushrooms |
| CN104072632A (en) * | 2014-07-15 | 2014-10-01 | 江苏阜丰生物科技有限公司 | Method for efficiently extracting maitake mycelia polysaccharides through submerged fermentation production |
| CN104387485A (en) * | 2014-11-12 | 2015-03-04 | 绥化学院 | Method for extracting polysaccharides in flammulina velutipes by synergism of complex enzymes and high-pressure hot water extraction process |
| CN107624513A (en) * | 2017-10-25 | 2018-01-26 | 山东惠民齐发果蔬有限责任公司 | It is a kind of rich in the edible and medical fungi cultural hypha method of polysaccharide and application |
| CN108294294A (en) * | 2018-01-19 | 2018-07-20 | 安徽省旌芝源畜业有限公司 | A kind of method that enzyme process prepares fat metabolism regulating composition |
| CN115381087A (en) * | 2022-06-22 | 2022-11-25 | 深圳市维龄可伴生物科技有限公司 | Efficient extraction process and application of needle mushroom powder |
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