CN1724685A - Method of extracting diosgenin by bioenzyme gradient catalysis - Google Patents
Method of extracting diosgenin by bioenzyme gradient catalysis Download PDFInfo
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- CN1724685A CN1724685A CN 200510014456 CN200510014456A CN1724685A CN 1724685 A CN1724685 A CN 1724685A CN 200510014456 CN200510014456 CN 200510014456 CN 200510014456 A CN200510014456 A CN 200510014456A CN 1724685 A CN1724685 A CN 1724685A
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Abstract
The invention discloses a method to distill yamogenin by using biology enzyme stage catalyzing that belongs to distill yamogenin from yam plant field. The method includes the following steps: adding shield-leaf yam or 'chuanlong' yam into biology reactor, and adding amylase, maltogenic amyiase, petinase, or emulsin, rough hesperidinase, maltogenic amyiase; or compounding cellulose, pentosan compounding enzyme and maltogenic amyiase processing for a while, taking stage catalyzing reaction; adding a certain consistency acid solution to take acid hydrolysis; washing to neutrality by basic salt solution; dipping for a certain while by non-polarity solution refluence; concentrating by evaporation, and crystallizing to gain product. The advantage of the invention is the distilling ratio of yamogenin could reach over 95%, low energy consumption and low producing cost.
Description
Technical field
The present invention relates to a kind of method of utilizing extracting diosgenin by bioenzyme gradient catalysis.Belong to the technology of extracting diosgenin by yam.
Background technology
Diosgenin is the starting raw material of preparation steroid hormone class medicine, is the important medicine intermediate that prepare steroid hormone class medicine and oral steroid contraceptive through eliminate acetic acid pregnant steroid diene alcohol ketone product that reaction back generate again by it through the product of cracking, oxidation, thus diosgenin to be called by the world of medicine be " medicinal gold ".
China plant worker begins to carry out the investigation and the development and use of Chinese yam resource in nineteen fifty-seven, and it is the steroid hormone pharmaceutical industries of main raw material with the diosgenin that beginning in 1958 has just been set up, and the later stage eighties begins the mass production diosgenin.At present, domestic have numerous manufacturer production steroid hormone class medicines and an oral steroid contraceptive, because the steroidal drug consumption is huge, also increases day by day as the demand of the diosgenin of raw material.In recent years, owing to excessively excavate, heavily gather, light protection makes the Chinese yam destruction Of resources very serious; Because wild Chinese yam is excavated the shortening in cycle, prolonged the resource recovery phase simultaneously, quality is reduced, and it is serious that germplasm is degenerated.Therefore the research of strengthening the extraction and separation technology to greatest extent of the artificial culture of Chinese yam resource and dioscin metamember is the important topic of pendulum in face of the various countries scientific worker.
It is best with Rhizome of Peltate Yam (Dioscorea zingiberensis Wight) and Dioscorea nipponica Mak. Ningpo Yam Rhizome (Dioscorea nipponica Makino) quality that China is used for producing the plant of diosgenin.The content of diosgenin approximately is respectively 2.5% and 1.5% in Rhizome of Peltate Yam and Dioscorea nipponica Mak. Ningpo Yam Rhizome rhizome.The method of traditional extraction diosgenin---directly acid-hydrolysis method is a method of using organic solvent extraction after the direct acid hydrolysis of employing again.This method is because the no thoroughness of hydrolysis, can only extract in the Chinese yam plant 50% sapogenin composition, and have problems such as energy consumption height, versatility be poor, seriously polluted, finally cause the production cost of diosgenin to improve, economic benefit is difficult to further improve.Someone has studied partition method and has extracted diosgenin, and this method is more perfect theoretically, but a large amount of foams makes separation be difficult for carrying out in the process of separating Chinese yam starch, and water-solubility saponin runs off easily, causes the yield of diosgenin not high.Many producers adopt pre-fermentation method to extract diosgenin both at home and abroad, carry out spontaneous fermentation exactly before acid hydrolysis earlier.But this method also only can extract in the Chinese yam about 60% diosgenin, and because because the uncertainty of fermentation condition causes the yield instability.The worker has studied the single influence that adds biological enzyme formulation (for example, cellulase, amylase etc.) to the diosgenin extract yield under pre-fermented inspiration both at home and abroad.Studies show that the adding of biological enzyme can impel dioscin in the Chinese yam plant to the conversion of diosgenin, thereby when extracting, can improve the yield of diosgenin.But owing to the complicacy of moiety in the Chinese yam and saponin(e structure, use single additional enzyme preparation specific aim not strong, limited the further raising of sapogenin yield.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing the catalysis of biological enzyme ladder to extract the diosgenin in the Chinese yam plant, the extraction yield that extracts diosgenin with this method from Chinese yam can reach more than 95%, and production cost is low.
The present invention is realized by following technical proposals.The method of the diosgenin in the Chinese yam plant is extracted in a kind of biological enzyme ladder catalysis, it is characterized in that comprising following process, with Rhizome of Peltate Yam, Dioscorea nipponica Mak. Ningpo Yam Rhizome is raw material, being ground into 20～50 orders joins in the bio-reactor, ml volumes ratio according to raw materials quality gram number and enzyme liquid is 1000: 1～1000: 3, in turn add amylase processing 1～3h that activity is 20000 μ/g at 60～80 ℃, add saccharifying enzyme processing 1～3h that activity is 100000 μ/g at 60～70 ℃, and add polygalacturonase processing 1～3h that activity are 15000 μ/g at 50～60 ℃; Perhaps in turn adding activity at 70～80 ℃ is that the synaptase of 10000 μ/g is handled 1～3h, and 60～70 ℃ add activity is that the thick hesperidinase of 5000 μ/g and saccharifying enzyme that activity is 100000 μ/g are handled 1～3h; Perhaps in turn add complex cellulase processing 1～3h that activity is 15000 μ/g at 40～50 ℃, 50～60 ℃ add piperylene prozyme processing 1～3h that activity is 2500 μ/g, and 60～70 ℃ add saccharifying enzyme processing 1～3h that activity is 100000 μ/g; Add the acid solution that concentration is acid solution 2～6mL of 1～5mol/L according to the 1g raw material then, and carry out acid hydrolysis 1～6h in 18～30 ℃; The subsalt solution washing of employing 2～5% is to neutral; Adopt non-polar solvent and be that 1: 15～60 quantity of solvent refluxes and leaches 4～10h by the ratio of hydrolysate quality and non-polar solvent volume; Final evaporation concentrates, and crystallization obtains product.
Above-mentioned acid solution is the aqueous solution of hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid.
The above-mentioned subsalt aqueous solution is the aqueous solution of yellow soda ash, salt of wormwood, sodium phosphate, sodium hydrogen phosphate or potassium hydrogen phosphate.
Above-mentioned non-polar solvent is normal hexane, sherwood oil, gasoline, butane, Skellysolve A or hexanaphthene.
The invention has the advantages that the katalysis of biological enzyme ladder can make that dioscin is converted into diosgenin to greatest extent in the plant, thereby make more than the extraction rate reached to 95% of diosgenin, and the catalytic production cycle of biological enzyme ladder can shorten to 1/10th of existing method, energy consumption is low, and production cost is low.
Embodiment
Embodiment one: the 50g Rhizome of Peltate Yam is ground into 40～50 orders places bio-reactor, add quantitative water and add the amylase processing 1h that the 0.05ml activity is 20000 μ/g down 60～80 ℃ of temperature; Attemperation is to add the saccharifying enzyme processing 2h that the 0.05ml activity is 100000 μ/g after 60～70 ℃; Regulate that to add the 0.05ml activity behind 50～60 ℃ of the feed temperatures be that the polygalacturonase of 15000 μ/g is handled 2h; Filtration is placed in the hydrochloric acid of quantitative 3mol/L, and carries out acid hydrolysis 5h in 25 ℃; Rhizome of Peltate Yam powder after the hydrolysis is extremely neutral with 3% aqueous sodium carbonate washing again, then in 50～60 ℃ of dryings; Dried hydrolyzate with the sherwood oil of the 300mL 6h that refluxes, is reclaimed sherwood oil with decompression evaporator and then with the product crystallization in extractor, dry diosgenin 1.25g in 80～100 ℃ of vacuum drying ovens at last, extraction yield can reach 95%.Product is through infrared measurement, result and the document (IR (KBr): v cm that matches
～11235,1044,975,912,895,862); Fusing point is 201 ℃; Results of elemental analyses is C:77.94%, and H:10.15% is consistent with calculated value (78.21% and 10.21%).
Embodiment two: the 100g Dioscorea nipponica Mak. Ningpo Yam Rhizome is ground into 20～30 orders places bio-reactor, add quantitative water and add the synaptase processing 2h that the 0.10ml activity is 10000 μ/g down 70～80 ℃ of temperature; Attemperation is to add the thick hesperidinase processing 2h that the 0.10ml activity is 5000 μ/g after 60～70 ℃; Attemperation is to add the saccharifying enzyme processing 2h that the 0.10ml activity is 100000 μ/g after 60～70 ℃; Filtration is placed in the sulfuric acid of 3mol/L of 400mL, and carries out acid hydrolysis 5h in 30 ℃; With 3% the sodium hydrogen phosphate aqueous solution that the hydrolyzate washing is extremely neutral then, then in 50～60 ℃ of dryings; 6h refluxes in extractor with the hexanaphthene of 800mL and Skellysolve A double solvents (volume proportion is 1: 1) with dried Dioscorea nipponica Mak. Ningpo Yam Rhizome powder, use the decompression evaporator concentrated extracting solution then and with the product crystallization, get diosgenin 1.35g at last in 80～100 ℃ of vacuum drying ovens after the drying, extraction yield can reach 96%.Product is through infrared measurement, result and the document (IR (KBr): vcm that matches
～11232,1046,980,921,896,859); Fusing point is 203 ℃; Results of elemental analyses is C:78.15%, and H:10.12% is consistent with calculated value (78.21% and 10.21%).
Embodiment three: the 50g Rhizome of Peltate Yam is ground into 20～30 orders places bio-reactor, add quantitative water and add the complex cellulase processing 2h that the 0.05ml activity is 15000 μ/g down 40～50 ℃ of temperature; Holding temperature is to add the piperylene prozyme processing 2h that the 0.05ml activity is 2500 μ/g after 50～60 ℃; Attemperation is to add the saccharifying enzyme processing 2h that the 0.05ml activity is 100000 μ/g after 60～70 ℃; Filtration is placed in the nitric acid of 3mol/L of 300mL, and carries out acid hydrolysis 5h in 25 ℃; With 3% sodium phosphate aqueous solution that the washing of Rhizome of Peltate Yam powder is extremely neutral then, then in 50～60 ℃ of dryings; Dried hydrolyzate with 120 gasoline of the 400mL 8h that refluxes, is reclaimed gasoline with decompression evaporator and then with the product crystallization in extractor, dry diosgenin 1.30g in 80～100 ℃ of vacuum drying ovens at last, extraction yield can reach 97%.Product is through infrared measurement, result and the document (IR (KBr): v cm that matches
～11240,1045,975,917.899,863); Fusing point is 204 ℃; Results of elemental analyses is C:79%, and H:10.01% is consistent with calculated value (78.21% and 10.21%).
Claims (4)
1. the method for an extracting diosgenin by bioenzyme gradient catalysis, it is characterized in that comprising following process, with Rhizome of Peltate Yam (Dioscorea zingiberensis Wight) and Dioscorea nipponica Mak. Ningpo Yam Rhizome (Dioscorea nipponica Makino) is raw material, being ground into 20～50 orders joins in the bio-reactor, ml volumes ratio according to raw materials quality gram number and enzyme liquid is 1000: 1～1000: 3, in turn add amylase processing 1～3h that activity is 20000 μ/g at 60～80 ℃, add saccharifying enzyme processing 1～3h that activity is 100000 μ/g at 60～70 ℃, and add polygalacturonase processing 1～3h that activity are 15000 μ/g at 50～60 ℃; Perhaps in turn adding activity at 70～80 ℃ is that the synaptase of 10000 μ/g is handled 1～3h, and 60～70 ℃ add activity is that the thick hesperidinase of 5000 μ/g and saccharifying enzyme that activity is 100000 μ/g are handled 1～3h; Perhaps in turn add complex cellulase processing 1～3h that activity is 15000 μ/g at 40～50 ℃, 50～60 ℃ add piperylene prozyme processing 1～3h that activity is 2500 μ/g, and 60～70 ℃ add saccharifying enzyme processing 1～3h that activity is 100000 μ/g; Add the acid solution that concentration is acid solution 2～6mL of 1～5mol/L according to the 1g raw material then, and carry out acid hydrolysis 1～6h in 18～30 ℃; The subsalt solution washing of employing 2～5% is to neutral; Adopt non-polar solvent and be that 1: 15～60 quantity of solvent refluxes and leaches 4～10h by the ratio of hydrolysate quality and non-polar solvent volume; Final evaporation concentrates, and crystallization obtains product.
2. extract the method for the diosgenin in the Chinese yam plant by the catalysis of the described biological enzyme ladder of claim 1, it is characterized in that acid solution is the aqueous solution of hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid.
3. extract the method for the diosgenin in the Chinese yam plant by the catalysis of the described biological enzyme ladder of claim 1, it is characterized in that the subsalt aqueous solution is the aqueous solution of yellow soda ash, salt of wormwood, sodium phosphate, sodium hydrogen phosphate or potassium hydrogen phosphate.
4. extract the method for the diosgenin in the Chinese yam plant by the catalysis of the described biological enzyme ladder of claim 1, it is characterized in that non-polar solvent is normal hexane, sherwood oil, gasoline, butane, Skellysolve A or hexanaphthene.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100439511C (en) * | 2006-11-07 | 2008-12-03 | 天津大学 | Process for catalytic extraction of yam saponin by using modified cellulase |
| CN100572553C (en) * | 2006-09-14 | 2009-12-23 | 中国科学院武汉植物园 | Method with preparing dioscin with dioscin penicillium notatum |
| CN101230378B (en) * | 2008-02-22 | 2011-08-10 | 西北农林科技大学 | Method for extracting axogenin by biological enzyme process |
| CN102336805A (en) * | 2011-01-07 | 2012-02-01 | 万绍平 | Method for extracting diosgenin from protodioscin |
| CN105238840A (en) * | 2015-10-27 | 2016-01-13 | 遵义市倍缘化工有限责任公司 | Method for preparing diosgenine by enzymatic hydrolysis |
| CN109651482A (en) * | 2019-01-23 | 2019-04-19 | 荆门市锦绣天成医药技术服务有限公司 | Furans steroid saponin in plant is transformed into the method for spiral steroid sapogenin |
-
2005
- 2005-07-11 CN CN 200510014456 patent/CN1724685A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100572553C (en) * | 2006-09-14 | 2009-12-23 | 中国科学院武汉植物园 | Method with preparing dioscin with dioscin penicillium notatum |
| CN100439511C (en) * | 2006-11-07 | 2008-12-03 | 天津大学 | Process for catalytic extraction of yam saponin by using modified cellulase |
| CN101230378B (en) * | 2008-02-22 | 2011-08-10 | 西北农林科技大学 | Method for extracting axogenin by biological enzyme process |
| CN102336805A (en) * | 2011-01-07 | 2012-02-01 | 万绍平 | Method for extracting diosgenin from protodioscin |
| CN102336805B (en) * | 2011-01-07 | 2013-03-27 | 万绍平 | Method for extracting diosgenin from protodioscin |
| CN105238840A (en) * | 2015-10-27 | 2016-01-13 | 遵义市倍缘化工有限责任公司 | Method for preparing diosgenine by enzymatic hydrolysis |
| CN109651482A (en) * | 2019-01-23 | 2019-04-19 | 荆门市锦绣天成医药技术服务有限公司 | Furans steroid saponin in plant is transformed into the method for spiral steroid sapogenin |
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