CN1262669C - Extraction of milk vetch polysaccharide by enzyme treatment - Google Patents
Extraction of milk vetch polysaccharide by enzyme treatment Download PDFInfo
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- CN1262669C CN1262669C CN 02158188 CN02158188A CN1262669C CN 1262669 C CN1262669 C CN 1262669C CN 02158188 CN02158188 CN 02158188 CN 02158188 A CN02158188 A CN 02158188A CN 1262669 C CN1262669 C CN 1262669C
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Abstract
The present invention relates to a method for extracting astragalus polysaccharide by enzyme treatment. Enzyme adopted by the present invention is one of the enzymes of cellulase, hemicellulase, proteinase, pectase and amylase, or a plurality of the enzymes of the cellulase, the hemicellulase, the proteinase, the pectase and the amylase, and can improve the extraction rate of the astragalus polysaccharide without damage to the structure of the astragalus polysaccharide. The extraction process of the present invention comprises the procedures of degreasing, enzyme treatment, leaching, alcohol deposition, drying, crushing, etc., and has the advantages of improved extraction rate without damage to the structure of the astragalus polysaccharide, improved content of the polysaccharide, mild conditions of the method for extracting the polysaccharide by the enzyme treatment, conformation maintaining of natural products of the astragalus polysaccharide, activity maintaining of effective ingredients and easy impurity removal.
Description
Technical field
The present invention relates to a kind of enzyme that utilizes and handle the method for extracting polysaccharide in the Radix Astragali.
Background technology:
The Radix Astragali (RadixAstragali) is the dry root of leguminous plants Radix Astagali Astragalus membranaceus (Fisch) Bge.Var.mongholicus (Bge) Hsiao. or Radix Astragali A.membranaceus (Fisch) Bge., belongs to tonic.Astragalus polysaccharides is one of main component of the Radix Astragali, and pharmacological action is widely arranged.
Traditional astragalus polysaccharides extracting method mainly be direct water carry or degreasing after water extraction, the medicinal part of the Radix Astragali is a root, wherein Mierocrystalline cellulose, hemicellulose and xylogen are all more, and the release to effective constituent in the leaching process has big inhibition, so extraction yield is on the low side.Though the extracting method that supersonic method or microwave method etc. are newer has certain effect to improving extraction yield, and is higher to equipment requirements, implements to acquire a certain degree of difficulty.Though acid or alkali extraction can improve extraction yield, acid can destroy the glycosidic bond of polysaccharide and form more monose and oligose, reduces astragalus polysaccharides content.Also can damage with the diluted alkaline extraction polysaccharide structures.In addition, need through neutralization after acid, the alkali lixiviate, program is loaded down with trivial details.
Mentioned that prozyme extracts the technology of polysaccharide protein in the mushroom among the Chinese patent CN 1110717A " polysaccharide protein and the capsular technology of manufacturing polysaccharide protein in the Enzymatic Extraction mushroom "; Mentioned the Enzymatic Extraction ginsenoside among the CN1144847A " enzyme is handled the method for extracting saponin component in the Chinese medicinal materials ".But do not see enzyme as yet and handle the report that extracts astragalus polysaccharides.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of enzyme that utilizes and handles the method for extracting astragalus polysaccharides.This method can improve the astragalus polysaccharides extraction yield, and does not destroy the astragalus polysaccharides structure.
The invention provides with enzyme and handle Radix Astragali raw material, extract the method for astragalus polysaccharides then.Connect because astragalus polysaccharides mostly is α-1,4, see that the selected enzyme of the present invention does not destroy functional astragalus polysaccharides and can destroy structural polysaccharide in the Radix Astragali raw material more and β-glycosidic link is comparatively in the structural polysaccharide.
Described enzyme is one or more combination in the following enzyme:
Cellulase, hemicellulase and proteolytic enzyme.
Described proteolytic enzyme is the enzyme of hydrolyzable vegetable-protein preferably, as papoid.
Extraction process of the present invention comprises steps such as degreasing, enzyme processing, lixiviate, alcohol precipitation, drying and pulverizing, and concrete operations are as follows:
1, degreasing
Radix Astragali raw material through the ethanolic soln degreasing that refluxes, is filtered, and filter residue volatilizes behind the solvent standby.
2, enzyme is handled
Step 1 gained filter residue was soaked in the aqueous solution that has added zymin 0.5-2 hour, selects pH value and temperature according to the suitable condition of selected zymin
3, extract
The heating said mixture filters, and collects filtrate, is the astragalus polysaccharides extracting solution.Treating processes with gained filter residue repeating step 2 and 3.
4, alcohol precipitation
Merge above-mentioned astragalus polysaccharides extracting solution, add ethanol and regulate, be settled out the astragalus polysaccharides solid.Dry this solid is pulverized, and obtains the astragalus polysaccharides crude product.
Enzyme is handled the process of extracting astragalus polysaccharides and mainly is made up of two stages.Fs is that the main effect of above-mentioned steps 2 is enzymolysis structure of cell surface and intercellular connector, and with the stripping of part glucide.Subordinate phase is after above-mentioned steps 3 is passed through to improve temperature, both to have had the enzyme of going out effect, makes thing leaching yield increase in the born of the same parents that dissolve in hot water simultaneously.
Advantage of the present invention is:
Improve extraction yield and do not destroy the astragalus polysaccharides structure, the extraction yield that can make polysaccharide can also improve the content of polysaccharide than not adopting enzyme treatment technique to improve more than the 20-50%.
Than traditional acid, alkali extraction and hot water extraction, enzyme handle to extract polysaccharide have mild condition, easily remove impurity, can keep the active advantage of astragalus polysaccharides.
The protein that the application of proteolytic enzyme not only can be decomposed in the Radix Astragali is that peptide or amino acid can also quicken the organization structure of the plant disintegration, makes the effectiveness performance of other enzyme better.
The present invention also is applicable to the extraction of all plant origin polyose effective constituents, also can improve the extraction yield of other soluble substance (as saponin, flavones etc.) simultaneously.
Embodiment
Further specify the present invention below in conjunction with embodiment, can not limit this but embodiment only is used for explanation
Scope of invention.
Embodiment 1
The A degreasing: with radix astragali coarse powder 1Kg, add 6 liters of ethanol, refluxing extraction 1.5 hours is filtered and is collected filter residue.
The B enzyme is handled: it is standby that above-mentioned filter residue is volatilized solvent, and in 8 premium on currency, adjust pH to 5.5 is with the filter residue mixing to add 2g hemicellulase (by streptomycete-tendril streptomycete (Streptomyces cirratus) self-control, the preparation method states as follows).50～60 ℃ of insulations, stirred 2 hours.
C is heated to little boiling with said mixture, extracts 2 hours, filters, and filtrate is polysaccharide extraction liquid A.
Filter residue is standby.
D adds 6 liters in water with step C gained filter residue, and little boiling extracted 1 hour, filtered, and filtrate is that polysaccharide is carried
Get liquid B.
Polysaccharide extraction liquid A and B that E combining step C and step D obtain, concentrating under reduced pressure, adding ethanol adjusting determining alcohol is 60～70%, placement is spent the night, to precipitate dry, pulverize astragalus polysaccharides crude product 109g, spectrophotometry polysaccharide content (phenolsulfuric acid method) is 26%.Compare yield with the control group that does not use enzyme to handle and improve 36%, content improves 40%.
The preparation method of above-mentioned hemicellulase is as follows:
1) medium component and content:
Plane and slant medium: corn cob xylan 1.4% peptone 0.1% yeast extract 0.1%
KH2PO4 0.1% MgSO4.H2O 0.05% agar 1.2% pH 5.7
Liquid fermentation medium: corn cob xylan 1.0%～2.0% peptone 1.4% yeast extract .03%
KH2PO4?1％ MgSO4.H2O?0.05％
2) preparation process
Is that slant medium is inoculated on the plating medium with streptomycete bacterial strain from preserving substratum, cultivates 5 days in 30 ℃ of incubators, select growth situation preferably bacterial strain 1～2 ring be inoculated in the liquid fermentation medium.The liquid fermentation and culture condition is: the liquid nutrient medium of the 50ml that in the triangular flask of 250ml, packs into, and with 140 rev/mins speed, 30 ℃ air constant temperature shaking table.In air constant temperature shaking table, cultivate after 5 days, with fermenting enzyme liquid with refrigerated centrifuge with 8000 rev/mins centrifugal 10 minutes, collect supernatant; Supernatant was dialysed in ice-water bath 12 hours, changed one time water in per 4 hours; Collect the interior liquid of dialysis tubing then and just obtain crude enzyme liquid.
Embodiment 2
The A degreasing: with radix astragali coarse powder 1Kg, add 4 liters of ethanol, refluxing extraction 1 hour is filtered and is collected filter residue.
The B enzyme is handled: it is standby that above-mentioned filter residue is volatilized solvent, and in 8 premium on currency, adjust pH to 6.7 is with the filter residue mixing to add 2g papoid (Papain zymetology numbering EC 3.4.22.2).60～70 ℃ of insulations, stirred 1 hour.
C is heated to little boiling with said mixture, extracts 1.5 hours, filters, and filtrate is polysaccharide extraction liquid A.Filter residue is standby.
D adds 4 liters in water with step C gained filter residue, adding commercially available cellulase (from aspergillus niger) and hemicellulase (is prepared by streptomycete-tendril streptomycete (Streptomyces cirratus), method is with embodiment 1) each 0.5g, adjust pH to 5.5, stirred 1 hour at 50～60 ℃ of insulations.Little boiling extracted 1 hour, filtered, and filtrate is polysaccharide extraction liquid B.
Polysaccharide extraction liquid A and B that E, combining step C and step D obtain, concentrating under reduced pressure, adding ethanol adjusting determining alcohol is 70～80%, placement is spent the night, to precipitate dry, pulverize astragalus polysaccharides crude product 113g, spectrophotometry polysaccharide content (phenolsulfuric acid method) is 28%.Compare yield with the control group that does not use enzyme to handle and improve 43%, content improves 49%.
Claims (3)
1, a kind of enzyme that utilizes is handled the method for extracting astragalus polysaccharides, comprises degreasing, enzyme processing, lixiviate, alcohol precipitation, drying and pulverising step, and concrete operations are as follows:
1) degreasing
Radix Astragali raw material through the ethanolic soln degreasing that refluxes, is filtered, and filter residue volatilizes behind the solvent standby;
2) enzyme is handled
Step 1 gained filter residue was soaked in the aqueous solution that has added zymin 0.5～2 hour; Described enzyme is one or more combination in the following enzyme: cellulase, hemicellulase and proteolytic enzyme;
3) extract
The heating said mixture filters, and collects filtrate, is the astragalus polysaccharides extracting solution; Treating processes with gained filter residue repeating step 2 and 3;
4) alcohol precipitation
Merge above-mentioned astragalus polysaccharides extracting solution, add ethanol and regulate, be settled out the astragalus polysaccharides solid; Dry this solid is pulverized, and obtains the astragalus polysaccharides crude product.
2, the described enzyme that utilizes of claim 1 is handled the method for extracting astragalus polysaccharides, and wherein said proteolytic enzyme is the enzyme of hydrolyzable vegetable-protein.
3, the described enzyme that utilizes of claim 2 is handled the method for extracting astragalus polysaccharides, and the enzyme of wherein said hydrolyzable vegetable-protein is a papoid.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02158188 CN1262669C (en) | 2002-12-24 | 2002-12-24 | Extraction of milk vetch polysaccharide by enzyme treatment |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02158188 CN1262669C (en) | 2002-12-24 | 2002-12-24 | Extraction of milk vetch polysaccharide by enzyme treatment |
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| CN1510143A CN1510143A (en) | 2004-07-07 |
| CN1262669C true CN1262669C (en) | 2006-07-05 |
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| CN 02158188 Expired - Fee Related CN1262669C (en) | 2002-12-24 | 2002-12-24 | Extraction of milk vetch polysaccharide by enzyme treatment |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966224A (en) * | 2010-10-20 | 2011-02-09 | 无锡正大畜禽有限公司 | Method for producing astragalus and codonopsis pilosula compound extract |
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| CN102875688A (en) * | 2012-10-05 | 2013-01-16 | 南通四海植物精华有限公司 | Process for extracting polysaccharide from milkvetch roots |
| CN102887963A (en) * | 2012-10-30 | 2013-01-23 | 河南牧翔动物药业有限公司 | Extraction method of astragalus polysaccharide |
| CN103382228A (en) * | 2013-07-31 | 2013-11-06 | 瑞普（天津）生物药业有限公司 | Preparation method for high-activity astragalus polysaccharide |
| CN103613674B (en) * | 2013-09-09 | 2015-12-02 | 华南农业大学 | A kind of extracting method of astragalus polysaccharides |
| CN105777921A (en) * | 2014-12-26 | 2016-07-20 | 王贵孝 | Astragalus polysaccharide extraction process and application thereof |
| CN105029154B (en) * | 2015-06-19 | 2018-08-14 | 恩施州源惠科技开发有限公司 | A kind of Siberian solomonseal rhizome polysaccharide rapidly dissolving tablet and processing method |
| CN105175573A (en) * | 2015-11-02 | 2015-12-23 | 黑龙江中医药大学 | High-efficiency radix astragali polysaccharide extraction method |
| CN106858598A (en) * | 2017-04-01 | 2017-06-20 | 吉林大学 | It is a kind of to specialize in micro-capsulized instant health food that puerpera eats and preparation method thereof |
| CN106977620A (en) * | 2017-06-03 | 2017-07-25 | 河南中医药大学 | A kind of method of purification of astragalus polyose |
| CN107267571A (en) * | 2017-07-15 | 2017-10-20 | 合肥市晶谷米业有限公司 | A kind of extracting method of rice bran polysaccharide |
| CN109576324A (en) * | 2018-11-15 | 2019-04-05 | 河北远征禾木药业有限公司 | A kind of astragalus polyose and its biological extraction method |
| CN110656130B (en) * | 2019-11-15 | 2021-05-11 | 滨州医学院 | Astragalus membranaceus-paecilomyces cicadae fermentation mycoplasm, preparation method and application |
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2002
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966224A (en) * | 2010-10-20 | 2011-02-09 | 无锡正大畜禽有限公司 | Method for producing astragalus and codonopsis pilosula compound extract |
| CN101966224B (en) * | 2010-10-20 | 2012-01-04 | 无锡正大畜禽有限公司 | Method for producing astragalus and codonopsis pilosula compound extract |
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| CN1510143A (en) | 2004-07-07 |
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