CN105112487A - Method for extracting dioscorea zingiberensis saponin by utilizing biological enzyme fermentation technology - Google Patents
Method for extracting dioscorea zingiberensis saponin by utilizing biological enzyme fermentation technology Download PDFInfo
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Abstract
The invention relates to a method for extracting dioscorea zingiberensis saponin by utilizing a biological enzyme fermentation technology. The method is characterized in that an enzyme needed during dioscorea zingiberensis fermentation is any one or a mixed enzyme of more of amylase, cellulase, hemicellulase, pectinase and protease. The total quantity of the added enzyme accounts for 1-10% (w/w) the weight of dioscorea zingiberensis. A certain proportion of biological enzyme is added during the dioscorea zingiberensis fermentation, the product yield can be improved remarkably, the fermentation time is shortened, and wastewater discharge is reduced. The method is convenient and practical and suitable for large-scale production and can meet ever-increasing market requirements.
Description
Technical field
The invention belongs to plant biological fermentation technical field, relate to a kind of method being extracted turmeric saponin by fermented by biological enzyme.
Technical background
Yellow ginger, formal name used at school Rhizome of Peltate Yam is Dioscoreaceae yam, is the distinctive wild plant resource of China.Containing abundant diosgenin in the root stock of yellow ginger, diosgenin is commonly called as diosgenin.Diosgenin is extensively present in yam, and wherein, in yellow ginger, the content of saponin is the highest.
In China, the utilization of people to yellow ginger has long history, edible, can be used as medicine, and is China's traditional Chinese medicine material.Saponin in yellow ginger has estrogen effect, cholesterol, cough-relieving can be reduced, eliminate the phlegm, desensitize, recover pathological tissues and stimulate heparinocyte and choleresis, its structure is through transformation or after modifying, also be widely used in the semi-synthetic raw material of steroidal anti-inflammatory medicine, contraceptive steroid, as cortisone, prednisone, training ground hundreds of different steroid hormone class medicines such as meter Song, and expensive, having the good reputation of " medicine gold ", is a kind of plant resources having very much potentiality to be exploited.
At present, the direct acid-hydrolysis method that the factory that China carries out saponin extraction generally adopts Rothrok pioneering.Working method is:
1, add in fermentor tank after yellow ginger (water content is 63% ~ 70%) being cleaned pulverizing, steam heating, interval passes into water vapor holding temperature at 40 DEG C, maintains this temperature fermentation 3 ~ 5 days;
2, moved in hydrolysis kettle by the yellow ginger after fermentation, add the hydrochloric acid soln of 10% ~ 15% of about 2 times amount (accounting for 2 times of yellow ginger input quantity, V/W), airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4% ~ 5%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, extract after 10h and lower the temperature, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling at 95 DEG C.
This reaction formula is:
Though this method is remote for duration of service, but still there are problems:
1, extract saponin yield in this way lower, product productive rate of giving money as a gift is only 1.5%, indirectly improves production cost.
2, wastewater discharge is huge, and space occupancy rate is large, high to the requirement of production unit.
Waste water in production of saponin mainly comes from fermentation: in a steam ambient, and 40 DEG C of bottom fermentations 3 ~ 5 days, can produce a large amount of water of condensation during this period, account for more than 50% of waste water total amount.With raw material charging capacity 150 tons note, product is about 0.83 ton, and wastewater discharge can reach 700 tons ~ 800 tons, and wherein, the waste water come from fermenting process about has 400 tons.And sewage disposal expense is expensive, treatment facility requires high.This is main, the severeest problem faced in current production of saponin.
Summary of the invention
In view of above-mentioned variety of problems, my company adopts biological enzyme technology, not only increases the yield of product, also decreases the quantity discharged of waste water in source, greatly reduce the processing costs of sewage, shorten the production cycle, reduce production cost.
For achieving the above object, the invention provides a kind of method utilizing fermented by biological enzyme yellow ginger to extract turmeric saponin, this invention overcomes many drawbacks of original technique, simple, can be used for business-like scale operation, meets the ever-increasing demand in market.Technology contents of the present invention is as follows:
Extract the method for turmeric saponin by biological enzyme technology fermentation yellow ginger, it is characterized in that the enzyme that will add when fermenting yellow ginger is the mixed enzyme of any one enzyme in amylase, cellulase, hemicellulase, polygalacturonase or proteolytic enzyme or multiple enzyme;
As optimization, the total amount of the enzyme added accounts for 1% ~ 10% (w/w) of yellow ginger weight.
As optimization, temperature during fermentation is at 20 DEG C ~ 40 DEG C, and fermentation time is 1 ~ 3 day.
The described method being extracted turmeric saponin by biological enzyme technology fermentation yellow ginger, comprises step:
(1) ferment: yellow ginger is cleaned, pulverizes, add the biological enzyme of 1% ~ 10% (w/w) accounting for yellow ginger weight, stir, sealing, 20 DEG C ~ 40 DEG C bottom fermentations 1 ~ 3 day;
(2) acidolysis: the yellow ginger after fermentation is added the hydrochloric acid soln of 10% ~ 15%, be warming up to 100 DEG C ~ 110 DEG C, hydrolysis 7.5-8.5 hour, cooling, standing, suction filtration;
(3) in filter residue, slowly add sodium hydroxide solution, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
(4) hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, extract after 10h and lower the temperature, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
(5) gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight; Stirring is warming up to backflow, and reflux about 2h; Lower the temperature afterwards, leave standstill, suction filtration, dry sterling at 95 DEG C.
As optimization, the bulking value proportioning of the yellow ginger described in step (2) and hydrochloric acid soln is 1:2.
As optimization, the concentration of sodium hydroxide solution 4% ~ 5% described in step (3).
The following content of high spot reviews of the present invention:
(1) kind of enzyme is on the impact of experimental result;
(2) consumption of enzyme is on the impact of experimental result;
(3) fermentation time of enzyme is on the impact of experimental result;
(4) temperature during enzymic fermentation is on the impact of experimental result;
Key problems-solving of the present invention is:
(1) the production of saponin cycle is long, and productive rate is low;
(2) in the production process of saponin, wastewater discharge is huge;
The positively effect compared with prior art had by the method for fermented by biological enzyme technology extraction turmeric saponin disclosed by the invention is:
1, product yield promotes greatly, and the production cycle shortens.
A. fermentation is a process allowing plant tissue fragmentation, adding a certain amount of enzyme, can crack plant tissue more completely sooner, effective constituent is come out sooner, more fully when fermenting.
Compare with the production result of same batch of yellow ginger, in traditional technology, 150 tons of yellow gingers produce the saponin of about 0.83 ton; After using novel process instead, 150 tons of yellow gingers can produce about 1.2 tons of saponin, and yield rose to 2.1% by 1.5% of the past.
B. adding enzyme when fermenting, effective constituent can be made to come out faster.The time of fermentation in past is 3 ~ 5 days, and even longer, using the novel process secondary fermentation time instead is 1 ~ 3 day, and cycle during production obviously shortens.
2, the output of waste water greatly reduces.Adopt steam heating in the past, and a large amount of water of condensation can be introduced during the fermentation, account for more than 50% of waste water total amount.In novel process, add enzyme after being cleaned and crush by yellow ginger, ferment in thermostatic equipment, new water of condensation can not be introduced in the process.The quantity discharged of sewage is greatly reduced.The waste water that past every batch produces is about 700 tons ~ 800 tons, and after improving, waste water output is 400 tons ~ 500 tons.Greatly reduce the processing costs of sewage, reduce production cost.
accompanying drawing illustrates:
Fig. 1 is the process flow sheet being extracted turmeric saponin method by fermented by biological enzyme technology.
embodiment:
Below in conjunction with specific embodiment, the present invention will be further described: following each embodiment is not only limitation of the present invention for illustration of the present invention.Agents useful for same of the present invention and conversion unit are commercially available prod, and biological enzyme formulation used is commercially available prod.
Embodiment 1:
The extraction of turmeric saponin
1, fermenting: 150g(water content is about 63%) yellow ginger cleans, pulverizes, add cellulase 3g, stir, sealing, 40 DEG C of bottom fermentations 3 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 10% hydrochloric acid soln 300ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 1.14g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.05%.
Embodiment 2:
The extraction of turmeric saponin
1, fermenting: 150g(water content is about 63%) yellow ginger cleans, pulverizes, add cellulase 7.5g, stir, sealing, 30 DEG C of bottom fermentations 1 day.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add the hydrochloric acid soln 300ml of 15%, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 1.17g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.11%.
Embodiment 3:
The extraction of turmeric saponin
1, fermenting: 600g(water content is about 63%) yellow ginger cleans, pulverizes, add hemicellulase 48g, stir, sealing, 35 DEG C of bottom fermentations 3 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 10% hydrochloric acid soln 1200ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 5%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 4.46g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.01%.
Embodiment 4:
The extraction of turmeric saponin
1, fermenting: 150g(water content is about 63%) yellow ginger cleans, pulverizes, add amylase 3g, stir, sealing, 30 DEG C of bottom fermentations 3 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add the hydrochloric acid soln 300ml of 10%, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 1.20g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.16%.
Embodiment 5:
The extraction of turmeric saponin
1, fermenting: 300g(water content is about 63%) yellow ginger cleans, pulverizes, add amylase 15g, stir, sealing, 36 DEG C of bottom fermentations 2 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 15% hydrochloric acid soln 600ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 2.32g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.09%.
Embodiment 6:
The extraction of turmeric saponin
1, fermenting: 600g(water content is about 63%) yellow ginger cleans, pulverizes, add polygalacturonase 48g, stir, sealing, 28 DEG C of bottom fermentations 2 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 15% hydrochloric acid soln 1200ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 5%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 4.55g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.05%.
Embodiment 7:
The extraction of turmeric saponin
1, fermenting: 150g(water content is about 63%) yellow ginger is cleaned, pulverizing, adds amylase 6g, cellulase 3g.Stir, sealing, 30 DEG C of bottom fermentations 1 day.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 10% hydrochloric acid soln 300ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 5%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 1.15g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.07%.
Embodiment 8:
The extraction of turmeric saponin
1, fermenting: 150g(water content is about 63%) yellow ginger is cleaned, pulverizing, adds amylase 3g, cellulase 6g.Stir, sealing, 20 DEG C of bottom fermentations 3 days.
2, acidolysis: moved in hydrolysis kettle by the yellow ginger after fermentation, add 10% hydrochloric acid soln 300ml, airtight, be warming up to 110 DEG C, be hydrolyzed about 8 hours, process discontinuous is exitted.Lower the temperature afterwards, leave standstill, suction filtration;
3, in filter residue, slowly add the sodium hydroxide solution of 4%, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
4, hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, lower the temperature after refluxing extraction 10h, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
5, gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight.Stirring is warming up to backflow, and reflux about 2h.Lower the temperature afterwards, leave standstill, suction filtration, dry sterling 1.12g at 95 DEG C.HPLC external standard method is surveyed content and is about 92%, and fusing point is 203 DEG C ~ 206 DEG C, and yield of giving money as a gift is 2.02%.
Embodiment 9
Simultaneous test:
Conclusion:
(1) method produces saponin to adopt the present invention to mention, saponin yield has obvious lifting, rose to 2.1%, and indirectly reduced production cost by 1.5% of the past.
(2) method produces saponin to adopt the present invention to mention, the quantity discharged of waste water reduces greatly, and 700 ~ 800 tons/batches by the past reduce to 400 ~ 500 tons/batches, reduce the pollution to environment.
Claims (6)
1. extract the method for turmeric saponin by biological enzyme technology fermentation yellow ginger, it is characterized in that the enzyme that will add when fermenting yellow ginger is the mixed enzyme of any one enzyme in amylase, cellulase, hemicellulase, polygalacturonase or proteolytic enzyme or multiple enzyme.
2. extracted the method for turmeric saponin described in claim 1 by biological enzyme technology fermentation yellow ginger, it is characterized in that the total amount of added enzyme accounts for 1% ~ 10% (w/w) of yellow ginger weight.
3. extract the method for turmeric saponin by biological enzyme technology fermentation yellow ginger described in claim 1, it is characterized in that temperature when fermenting is at 20 DEG C ~ 40 DEG C, fermentation time is 1 ~ 3 day.
4. the method being extracted turmeric saponin by biological enzyme technology fermentation yellow ginger according to claim 1, be is characterized in that, comprise step:
(1) ferment: yellow ginger is cleaned, pulverizes, add the biological enzyme of 1% ~ 10% (w/w) accounting for yellow ginger weight, stir, sealing, 20 DEG C ~ 40 DEG C bottom fermentations 1 ~ 3 day;
(2) acidolysis: the yellow ginger after fermentation is added the hydrochloric acid soln of 10% ~ 15%, be warming up to 100 DEG C ~ 110 DEG C, hydrolysis 7.5-8.5 hour, cooling, standing, suction filtration;
(3) in filter residue, slowly add sodium hydroxide solution, adjust ph is 7, standing, suction filtration, and gained hydrolyzate is dry at 95 DEG C, obtains dry hydrolyzate;
(4) hydrolyzate is dropped in extractor, adds white spirit, stir and heat up, extract after 10h and lower the temperature, after concentrated, distillation solid distillment, dry saponin crude product at 95 DEG C;
(5) gained crude product is dropped into recrystallization in sherwood oil, sherwood oil consumption is 2 ~ 3 times (V/W) of crude product weight; Stirring is warming up to backflow, and reflux about 2h; Lower the temperature afterwards, leave standstill, suction filtration, dry sterling at 95 DEG C.
5. the method being extracted turmeric saponin by biological enzyme technology fermentation yellow ginger according to claim 4, it is characterized in that, the bulking value proportioning of the yellow ginger described in step (2) and hydrochloric acid soln is 1:2.
6. the method being extracted turmeric saponin by biological enzyme technology fermentation yellow ginger according to claim 4, be is characterized in that, the concentration of sodium hydroxide solution 4% ~ 5% described in step (3).
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106046108A (en) * | 2016-04-01 | 2016-10-26 | 合肥工业大学 | Novel process for extracting diosgenine through fermentation of yellow ginger via Mortierella elongate PFY |
| CN106086150A (en) * | 2016-07-25 | 2016-11-09 | 陕西科技大学 | A kind of method utilizing enzyme to combine microorganism conversion production turmeric saponin |
| CN107522767A (en) * | 2017-10-13 | 2017-12-29 | 陈增光 | A kind of preparation method of turmeric saponin |
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| CN1544649A (en) * | 2003-11-11 | 2004-11-10 | 姜中兴 | Process for extracting saponin |
| CN101397582A (en) * | 2007-09-30 | 2009-04-01 | 咸阳宏华生化科技有限公司 | Method for producing diosgenin by environment friendly pollution-free holoenzyme method |
| CN101857855A (en) * | 2010-07-05 | 2010-10-13 | 沅江浣溪沙酶技术有限公司 | Complex enzyme preparation and application in producing diosgenin |
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| CN101397582A (en) * | 2007-09-30 | 2009-04-01 | 咸阳宏华生化科技有限公司 | Method for producing diosgenin by environment friendly pollution-free holoenzyme method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106046108A (en) * | 2016-04-01 | 2016-10-26 | 合肥工业大学 | Novel process for extracting diosgenine through fermentation of yellow ginger via Mortierella elongate PFY |
| CN106086150A (en) * | 2016-07-25 | 2016-11-09 | 陕西科技大学 | A kind of method utilizing enzyme to combine microorganism conversion production turmeric saponin |
| CN107522767A (en) * | 2017-10-13 | 2017-12-29 | 陈增光 | A kind of preparation method of turmeric saponin |
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