CN107987178A - A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide - Google Patents
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide Download PDFInfo
- Publication number
- CN107987178A CN107987178A CN201711246490.3A CN201711246490A CN107987178A CN 107987178 A CN107987178 A CN 107987178A CN 201711246490 A CN201711246490 A CN 201711246490A CN 107987178 A CN107987178 A CN 107987178A
- Authority
- CN
- China
- Prior art keywords
- radix isatidis
- parts
- polysaccharide
- mass fraction
- soak
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/04—Plant cells or tissues
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Sustainable Development (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to the extracting method of technical field of traditional Chinese medicines, more particularly to a kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide.The present invention carries out the pulverization process of Radix Isatidis using air-flow crushing, Radix Isatidis structure can more be refined, extraction polysaccharide will not be impacted, by the mechanical support wall membrane structure for destroying Radix Isatidis, later stage can make the more complete of polysaccharide dissolution, steam blasting carries out pre-treatment, using high temperature saturated vapor rapid osmotic into Radix Isatidis cell tissue, and component separation and the structure change of raw material are realized by moment pressure release, so as to destroy the cell tissue of Radix Isatidis, destroy the interconnection between fibre bundle, make cell wall rupture, form loose and porous structure, fibre bundle is separated from each other, cell wall rupture, so that being wrapped in fibre bundle or intracellular functional components are released, significantly reduce resistance to mass tranfer, achieve the purpose that to improve polysaccharide extract rate.
Description
Technical field
The present invention relates to the extracting method of technical field of traditional Chinese medicines, more particularly to a kind of it is used for the extraction Banlangen Polysaccharide that ferments
Radix Isatidis preprocess method.
Background technology
Radix Isatidis, alias indigo-blue, indigo root, indigo root, be cruciferae isatis dry root, there is heat-clearing solution
The effect of poison, cool blood relieving sore-throat, be used clinically for poison during warm epidemic disease, fever pharyngalgia, febrile virulent maculae, mumps, scarlet fever, infection with swollen head
Epidemic disease, erysipelas, carbuncle swells etc..As the Typical Representative of China's anti virus herb, Radix Isatidis is first to obtain National Institutes of Health
The traditional Chinese medicine research project of subsidy.
Polysaccharide is a kind of baroque large biological molecule being formed by connecting by multiple monosaccharide molecules by glycosidic bond, natural
It is distributed in medicine very extensive.Polysaccharide biosynthesis and intercellular identification, fertilization, embry ogenesis, neural cell development,
Played in many basic life processes such as hormone activation, cell Proliferation, virus and the infection of bacterium, Nasopharyngeal neoplasms important
Effect, therefore become the research hotspot of medicine, nutrition and field of food science.It is reported that Banlangen Polysaccharide is to specificity
Immune and nospecific immunity is respectively provided with certain facilitation, is a kind of more satisfactory innate immunity reinforcing agent, as guarantor
Health food and medicine have extensive development prospect.
Banlangen Polysaccharide has antivirus action, has certain humidification to nospecific immunity and humoral immunity.
Its crude polysaccharide extract contains the impurity such as a large amount of protein, pigment, and the purity and drug effect to polyose have considerable influence, especially
It is that it is one essential to remove albumen contained in Thick many candies and pigment when being developed into the higher medicine of purity
Purification step.
Traditional polysaccharide method for removing protein has Seavag methods, trichloroacetic acid method, HCl methods, enzyme process etc., wherein Seavag methods most
It is common, Seavag methods are that trichloroacetic acid and n-butanol are mixed in a certain ratio and polysaccharide liquid extracted, and are repeated several times, until
Take off albumen.Using the method, not only polysaccharide loss amount is big, and the organic solvent in product is difficult to cleared；And organic solvent usage amount
Greatly, of high cost, pollution environment.The active charcoal absorptionization of polysaccharide discoloration method, macroreticular resin absorbing method, oxidizing process etc..Use with
Upper method, usual decolorizing efficiency is not high, or necessarily loss can be caused to polysaccharide, is destroyed.In traditional handicraft, it is normal with decoloration to take off albumen
Complete in two steps, complex process, time-consuming.
The method of traditional extraction Banlangen Polysaccharide is mainly water extraction and alcohol precipitation method, this method also by《Chinese Pharmacopoeia》Receive
Carry, mainly using thermophilic digestion, ethanol precipitation.This method is not required to special installation, but very long with the operating time, and energy consumption is big, has
The shortcomings of effect component high temperature easily decomposes, and recovery rate is very low, and the dregs of a decoction after extraction are not disposable, it is clear that inadaptable industry metaplasia
Production.The hot fields of research are become using probiotics fermention Chinese herbal medicine in recent years, at present lactic acid bacteria, Bacillus, yeast
The report of bacterium fermentation of Chinese herbal medicine.Research shows that microorganism has very abundant enzyme system, has powerful decomposition and inversion material
Ability, after Chinese herbal medicine is fermented, its active ingredient and active material are discharged to greatest extent, while can also be produced
New active material.But these method fermentation times are longer, process is more extensive, and recovery rate is not high, and it is even more impossible to carry out industry
Metaplasia is produced.
In summary, the preparation method of present Banlangen Polysaccharide, it is overall there is it is following the defects of：It is easy to cause more
The loss late of sugar is excessive, and extraction efficiency is too low, environment easy to pollute, it is impossible to as large-scale industrial production mode.
Therefore the high Banlangen Polysaccharide extracting method of a kind of high income, purity how is obtained, is those skilled in the art
Technical problem urgently to be resolved hurrily.
The content of the invention
The technical problems to be solved by the invention：Asked for current microbial fermentation Banlangen Polysaccharide recovery rate is not high
A kind of topic, there is provided Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide.
In order to solve the above technical problems, the present invention is using technical solution as described below：
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, this method comprises the following steps：
（1）The lateral bud of internode is taken to be placed in the ethanol solution that mass fraction is 70% and soak as explant from Radix Isatidis plant
Bubble, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak, taking-up rinsed with sterile water, is seeded in plant culture
Illumination cultivation, it is to be soaked in 0.01mol/L citric acid solutions to take culture back plate indigo plant root explant to add activated carbon and concentration, must be soaked
Radix Isatidis after bubble；
（2）Radix Isatidis takes dried object to be placed in air-flow crushing, crosses 100 mesh sieves, collect sieving in 40 ~ 50 DEG C of dryings after taking immersion
Grain, takes sieving particle to be placed in 55 ~ 60 DEG C of constant weights, takes constant weight sieving particle Steam explosion treatment, Radix Isatidis after must handling；
（3）According to the mass fraction, 7 ~ 10 parts of rhamnolipid is taken, 20 ~ 25 parts of cellulase, 5 ~ 8 parts of phenylalanine, polyethylene glycol 5 ~
8 parts, mass fraction is 70% 60 ~ 90 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, Radix Isatidis adds leaching after taking processing
Soaked in bubble liquid, take out Radix Isatidis, it is dry, obtain the pretreatment Radix Isatidis for the extraction Banlangen Polysaccharide that ferments；
The step（1）Middle plant culture：According to the mass fraction, 60 ~ 70 parts of ammonium nitrate, 100 ~ 110 parts of potassium nitrate, phosphoric acid are taken
120 ~ 130 parts of potassium dihydrogen, 200 ~ 250 parts of magnesium sulfate, 140 ~ 150 parts of calcium chloride, 6-BA0.02 ~ 0.06 part, methyl α-naphthyl acetate 0.02 ~
0.05 part, 20 ~ 30 parts of sucrose, 20 ~ 25 parts of agar, 400 ~ 500 parts of water, obtains plant culture.
The step（1）Middle Radix Isatidis explant, which is placed in the ethanol solution that mass fraction is 70%, soaks 30 ~ 40s, is placed in
5 ~ 6min is soaked in the mercuric chloride solution that mass fraction is 0.1%；Illumination cultivation condition：23 ~ 27 DEG C of 5 ~ 7d of culture, intensity of illumination
For 2000lx, daily light application time is 12h；It is 0.01mol/L citric acids that back plate indigo plant root explant, which is cultivated, with activated carbon and concentration
The mass ratio of solution is 10：1：0.3 ~ 0.5, soak 2 ~ 2.5h.
The step（2）Middle steam blasting condition is：150MPa, pressurize 150s.
The step（3）The mass ratio of Radix Isatidis and soak is 1 after middle processing：5~7；30 ~ 35 DEG C of 30 ~ 40min of immersion.
Compared with other methods, advantageous effects are the present invention：
（1）The present invention carries out tissue cultures, on the one hand can quickly be bred using Radix Isatidis explant is cut, and shortens plate
The time of blue root cells propagation, variation is reduced, quality is kept, meets the needs of market is to Radix Isatidis, on the other hand leave parent
Radix Isatidis incision easily brown stain afterwards, influences the activity of later stage extraction Banlangen Polysaccharide, and prevent-browning is carried out to Radix Isatidis notch
Processing, protection incision membrane structure and the destruction for mitigating Physical Zoneization distribution in cell, it is therefore prevented that for extracting the work of polysaccharide
Property material exudation tissue, enhance cytoactive, improve later stage fermentation extraction polysaccharide efficiency；
（2）The present invention carries out the pulverization process of Radix Isatidis using air-flow crushing, can more refine Radix Isatidis structure, will not be to carrying
Polysaccharide is taken to impact, by destroying the mechanical support wall membrane structure of Radix Isatidis, the later stage can make the more complete of polysaccharide dissolution,
Steam blasting carries out pre-treatment, using high temperature saturated vapor rapid osmotic into Radix Isatidis cell tissue, and passes through moment pressure release
Realize component separation and the structure change of raw material, so as to destroy the cell tissue of Radix Isatidis, destroy the mutually interconnection between fibre bundle
Connect, make cell wall rupture, form loose and porous structure, fibre bundle is separated from each other, cell wall rupture so that be wrapped in fibre bundle or
Intracellular functional components are released, hence it is evident that are reduced resistance to mass tranfer, achieveed the purpose that to improve polysaccharide extract rate；
（3）After steam blasting, iuntercellular hole is opened, takes cellulase to carry out compounding and soak is made, by Radix Isatidis
It is soaked in soak, the active ingredient of soak enters hole, and cellulase can improve the content of soluble sugar, cellulose
Enzyme is diffused into intramolecule hydrolysis crystalline cellulose, the function influence substrate structure between polyethylene glycol and substrate, and reduction finishes
The reaction activity of crystalline region, can be to avoid the ineffective adsorption of enzyme, and phenylalanine and rhamnolipid can also promote the activity of enzyme, reduce
The loss of enzyme, improves the utilization rate of enzyme, makes the more dissolutions of energy under the catalysis of enzyme of later stage fermentation active material, improves polysaccharide and carry
Take rate.
Embodiment
Plant culture：According to the mass fraction, 60 ~ 70 parts of ammonium nitrate, 100 ~ 110 parts of potassium nitrate, potassium dihydrogen phosphate 120 are taken
~ 130 parts, 200 ~ 250 parts of magnesium sulfate, 140 ~ 150 parts of calcium chloride, 6-BA0.02 ~ 0.06 part, 0.02 ~ 0.05 part of methyl α-naphthyl acetate, sucrose
20 ~ 30 parts, 20 ~ 25 parts of agar, 400 ~ 500 parts of water, obtains plant culture.
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, includes the following steps：
（1）The lateral bud that internode is taken from Radix Isatidis plant is explant, is placed in the ethanol solution that mass fraction is 70% and soaks
30 ~ 40s, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak 5 ~ 6min, take out Radix Isatidis rinsed with sterile water 3 ~ 4
It is secondary, it is seeded in plant culture, 23 ~ 27 DEG C of 5 ~ 7d of culture, intensity of illumination 2000lx, daily light application time is 12h, takes training
Support back plate indigo plant root explant in mass ratio 10：1：0.3 ~ 0.5 addition activated carbon and concentration are to be soaked in 0.01mol/L citric acid solutions
Steep 2 ~ 2.5h, Radix Isatidis after must soaking；
（2）Radix Isatidis takes dried object air-flow crushing, sets 3 kg/h of charging rate, work pressure in 40 ~ 50 DEG C of dryings after taking immersion
0.8 MPa of power, equipment tolerance 2m3/ min, crosses 100 mesh sieves, collects sieving particle, takes sieving particle to be placed in 55 ~ 60 DEG C of constant weights, takes
Constant weight sieving particle be placed in the processing chamber of high density steam blasting machine, then pass to saturated vapor, set vapour pressure as
150MPa, pressurize 150s processing, Radix Isatidis after must handling；
（3）According to the mass fraction, 7 ~ 10 parts of rhamnolipid is taken, 20 ~ 25 parts of cellulase, 5 ~ 8 parts of phenylalanine, polyethylene glycol 5 ~
8 parts, mass fraction is 70% 60 ~ 90 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, Radix Isatidis is by quality after taking processing
Than 1：5 ~ 7 add in soak in 30 ~ 35 DEG C of 30 ~ 40min of immersion, take out Radix Isatidis, dry, obtain and extract Radix Isatidis for fermenting
The pretreatment Radix Isatidis of polysaccharide；
（4）Bacillus subtilis is taken to be inoculated in nutrient broth medium, 33 ~ 37 DEG C of 1 ~ 2d of culture, take saccharomyces cerevisiae to be inoculated in
In malt extract medium, 35 ~ 37 DEG C of 1 ~ 2d of culture, take bacillus subtilis culture in mass ratio 2：1 mixing, obtains Mixed Microbes
Liquid, according to the mass fraction, takes 80 ~ 85 parts of wheat bran, and 9 ~ 15 parts of bean cake powder, 10 ~ 12 parts of corn flour, is made fermentation substrate, take in
The pretreatment Radix Isatidis and mixed bacteria liquid and fermentation substrate in mass ratio 20 of fermentation extraction Banlangen Polysaccharide：3：50 ~ 60 mixing, in
27 ~ 37 DEG C of 3 ~ 5d of fermentation, obtain fermentate, centrifuge, take supernatant in mass ratio 1：9 add the ethanol that mass fraction is 95%, stand
3 ~ 5h, centrifugation, takes precipitation to be washed 2 ~ 3 times with the ethanol solution of mass fraction 80%, is freeze-dried, measures carrying for Banlangen Polysaccharide
Take rate to reach 45 ~ 47%, take unprocessed Radix Isatidis to repeat microbial fermentation, extraction process, measures the recovery rate of Banlangen Polysaccharide
For 23 ~ 27%.
Embodiment 1
Plant culture：According to the mass fraction, 60 parts of ammonium nitrate, 100 parts of potassium nitrate, 120 parts of potassium dihydrogen phosphate, magnesium sulfate are taken
200 parts, 140 parts of calcium chloride, 6-BA0.02 parts, 0.02 part of methyl α-naphthyl acetate, 20 parts of sucrose, 20 parts of agar, 400 parts of water, obtains plant training
Support base.
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, includes the following steps：
（1）The lateral bud that internode is taken from Radix Isatidis plant is explant, is placed in the ethanol solution that mass fraction is 70% and soaks
30s, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak 5min, takes out Radix Isatidis rinsed with sterile water 3 times, inoculation
Into plant culture, 23 DEG C of culture 5d, intensity of illumination 2000lx, daily light application time is 12h, is taken after cultivating outside Radix Isatidis
Implant in mass ratio 10：1：0.3 addition activated carbon and concentration are to soak 2h in 0.01mol/L citric acid solutions, obtain immersion back plate
Lan Gen；
（2）Radix Isatidis takes dried object air-flow crushing, sets 3 kg/h of charging rate, operating pressure in 40 DEG C of dryings after taking immersion
0.8 MPa, equipment tolerance 2m3/ min, crosses 100 mesh sieves, collects sieving particle, takes sieving particle to be placed in 55 DEG C of constant weights, takes constant weight
Sieving particle is placed in the processing chamber of high density steam blasting machine, then passes to saturated vapor, sets vapour pressure as 150MPa, guarantor
Press 150s processing, Radix Isatidis after must handling；
（3）According to the mass fraction, 7 parts of rhamnolipid, 20 parts of cellulase, 5 parts of phenylalanine, 5 parts of polyethylene glycol, quality point are taken
Number is 70% 60 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, take Radix Isatidis in mass ratio 1 after processing：5 add immersion
30min is soaked in 30 DEG C in liquid, takes out Radix Isatidis, it is dry, obtain the pretreatment Radix Isatidis for the extraction Banlangen Polysaccharide that ferments；
（4）Bacillus subtilis is taken to be inoculated in nutrient broth medium, 33 DEG C of culture 1d, take saccharomyces cerevisiae to be inoculated in brewer's wort
In culture medium, 35 DEG C of culture 1d, take bacillus subtilis culture in mass ratio 2：1 mixing, obtains mixed bacteria liquid, by mass fraction
Meter, takes 80 parts of wheat bran, and 9 parts of bean cake powder, 10 parts of corn flour, is made fermentation substrate, takes in the pre- of fermentation extraction Banlangen Polysaccharide
Handle Radix Isatidis and mixed bacteria liquid and fermentation substrate in mass ratio 20：3：50 mixing, ferment 3d in 27 DEG C, obtains fermentate, centrifuges,
Take supernatant in mass ratio 1：9 add the ethanol that mass fraction is 95%, stand 3h, centrifugation, takes precipitation mass fraction 80%
Ethanol solution washs 2 times, freeze-drying, measures the extraction rate reached of Banlangen Polysaccharide to 45%, takes unprocessed Radix Isatidis to repeat micro-
Biofermentation, extraction process, the recovery rate for measuring Banlangen Polysaccharide are 23%.
Embodiment 2
Plant culture：According to the mass fraction, 65 parts of ammonium nitrate, 105 parts of potassium nitrate, 125 parts of potassium dihydrogen phosphate, magnesium sulfate are taken
225 parts, 145 parts of calcium chloride, 6-BA0.04 parts, 0.04 part of methyl α-naphthyl acetate, 25 parts of sucrose, 23 parts of agar, 450 parts of water, obtains plant training
Support base.
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, includes the following steps：
（1）The lateral bud that internode is taken from Radix Isatidis plant is explant, is placed in the ethanol solution that mass fraction is 70% and soaks
35s, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak 5min, takes out Radix Isatidis rinsed with sterile water 3 times, inoculation
Into plant culture, 24 DEG C of culture 6d, intensity of illumination 2000lx, daily light application time is 12h, is taken after cultivating outside Radix Isatidis
Implant in mass ratio 10：1：0.4 addition activated carbon and concentration are to soak 2.4h in 0.01mol/L citric acid solutions, after obtaining immersion
Radix Isatidis；
（2）Radix Isatidis takes dried object air-flow crushing, sets 3 kg/h of charging rate, operating pressure in 45 DEG C of dryings after taking immersion
0.8 MPa, equipment tolerance 2m3/ min, crosses 100 mesh sieves, collects sieving particle, takes sieving particle to be placed in 57 DEG C of constant weights, takes constant weight
Sieving particle is placed in the processing chamber of high density steam blasting machine, then passes to saturated vapor, sets vapour pressure as 150MPa, guarantor
Press 150s processing, Radix Isatidis after must handling；
（3）According to the mass fraction, 8 parts of rhamnolipid, 23 parts of cellulase, 7 parts of phenylalanine, 7 parts of polyethylene glycol, quality point are taken
Number is 70% 70 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, take Radix Isatidis in mass ratio 1 after processing：6 add immersion
35min is soaked in 33 DEG C in liquid, takes out Radix Isatidis, it is dry, obtain the pretreatment Radix Isatidis for the extraction Banlangen Polysaccharide that ferments；
（4）Bacillus subtilis is taken to be inoculated in nutrient broth medium, 34 DEG C of culture 1d, take saccharomyces cerevisiae to be inoculated in brewer's wort
In culture medium, 36 DEG C of culture 1d, take bacillus subtilis culture in mass ratio 2：1 mixing, obtains mixed bacteria liquid, by mass fraction
Meter, takes 83 parts of wheat bran, and 11 parts of bean cake powder, 11 parts of corn flour, is made fermentation substrate, takes in the pre- of fermentation extraction Banlangen Polysaccharide
Handle Radix Isatidis and mixed bacteria liquid and fermentation substrate in mass ratio 20：3：55 mixing, ferment 4d in 35 DEG C, obtains fermentate, centrifuges,
Take supernatant in mass ratio 1：9 add the ethanol that mass fraction is 95%, stand 4h, centrifugation, takes precipitation mass fraction 80%
Ethanol solution washs 2 times, freeze-drying, measures the extraction rate reached of Banlangen Polysaccharide to 46%, takes unprocessed Radix Isatidis to repeat micro-
Biofermentation, extraction process, the recovery rate for measuring Banlangen Polysaccharide are 24%.
Embodiment 3
Plant culture：According to the mass fraction, 70 parts of ammonium nitrate, 110 parts of potassium nitrate, 130 parts of potassium dihydrogen phosphate, magnesium sulfate are taken
250 parts, 150 parts of calcium chloride, 6-BA0.06 parts, 0.05 part of methyl α-naphthyl acetate, 20 ~ 30 parts of sucrose, 20 ~ 25 parts of agar, water 400 ~ 500
Part, obtain plant culture.
A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, includes the following steps：
（1）The lateral bud that internode is taken from Radix Isatidis plant is explant, is placed in the ethanol solution that mass fraction is 70% and soaks
40s, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak 6min, takes out Radix Isatidis rinsed with sterile water 4 times, inoculation
Into plant culture, 27 DEG C of culture 7d, intensity of illumination 2000lx, daily light application time is 12h, is taken after cultivating outside Radix Isatidis
Implant in mass ratio 10：1：0.5 addition activated carbon and concentration are to soak 2.5h in 0.01mol/L citric acid solutions, after obtaining immersion
Radix Isatidis；
（2）Radix Isatidis takes dried object air-flow crushing, sets 3 kg/h of charging rate, operating pressure in 50 DEG C of dryings after taking immersion
0.8 MPa, equipment tolerance 2m3/ min, crosses 100 mesh sieves, collects sieving particle, takes sieving particle to be placed in 60 DEG C of constant weights, takes constant weight
Sieving particle is placed in the processing chamber of high density steam blasting machine, then passes to saturated vapor, sets vapour pressure as 150MPa, guarantor
Press 150s processing, Radix Isatidis after must handling；
（3）According to the mass fraction, 10 parts of rhamnolipid, 25 parts of cellulase, 8 parts of phenylalanine, 8 parts of polyethylene glycol, quality are taken
Fraction is 70% 90 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, take Radix Isatidis in mass ratio 1 after processing：7 add leaching
Steep in liquid and soak 40min in 35 DEG C, take out Radix Isatidis, it is dry, obtain the pretreatment Radix Isatidis for the extraction Banlangen Polysaccharide that ferments；
（4）Bacillus subtilis is taken to be inoculated in nutrient broth medium, 37 DEG C of culture 2d, take saccharomyces cerevisiae to be inoculated in brewer's wort
In culture medium, 37 DEG C of culture 2d, take bacillus subtilis culture in mass ratio 2：1 mixing, obtains mixed bacteria liquid, by mass fraction
Meter, takes 85 parts of wheat bran, and 15 parts of bean cake powder, 12 parts of corn flour, is made fermentation substrate, takes in the pre- of fermentation extraction Banlangen Polysaccharide
Handle Radix Isatidis and mixed bacteria liquid and fermentation substrate in mass ratio 20：3：60 mixing, ferment 5d in 37 DEG C, obtains fermentate, centrifuges,
Take supernatant in mass ratio 1：9 add the ethanol that mass fraction is 95%, stand 3 ~ 5h, centrifugation, takes precipitation mass fraction 80%
Ethanol solution wash 3 times, freeze-drying, measures the extraction rate reached of Banlangen Polysaccharide to 47%, takes unprocessed Radix Isatidis to repeat
Microbial fermentation, extraction process, the recovery rate for measuring Banlangen Polysaccharide are 27%.
Comparative example
Bacillus subtilis is taken to be inoculated in nutrient broth medium, 37 DEG C of culture 2d, take saccharomyces cerevisiae to be inoculated in brewer's wort training
Support in base, 37 DEG C of culture 2d, take bacillus subtilis culture in mass ratio 2：1 mixing, obtains mixed bacteria liquid, by mass fraction
Meter, takes 85 parts of wheat bran, and 15 parts of bean cake powder, 12 parts of corn flour, is made fermentation substrate, take untreated Radix Isatidis and mixed bacteria liquid and
Fermentation substrate in mass ratio 20：3：60 mixing, ferment 5d in 37 DEG C, obtains fermentate, centrifuges, take supernatant in mass ratio 1：9 add
Enter the ethanol that mass fraction is 95%, stand 3 ~ 5h, centrifugation, takes precipitation to be washed 3 times with the ethanol solution of mass fraction 80%, freezes
It is dry, the extraction rate reached of Banlangen Polysaccharide is measured to 35%.
From the foregoing, it will be observed that the Radix Isatidis preprocess method for being used to extract Banlangen Polysaccharide that the present invention is invented is for Radix Isatidis
The recovery rate of polysaccharide is higher, and the method for the present invention is a kind of safe and efficient processing method, is worthy to be popularized and uses.
Claims (5)
1. a kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide, it is characterised in that this method includes as follows
Step：
（1）The lateral bud of internode is taken to be placed in the ethanol solution that mass fraction is 70% and soak as explant from Radix Isatidis plant
Bubble, then be placed in the mercuric chloride solution that mass fraction is 0.1% and soak, Radix Isatidis rinsed with sterile water is taken out, is seeded to plant training
Illumination cultivation in base is supported, it is to be soaked in 0.01mol/L citric acid solutions to take culture back plate indigo plant root explant to add activated carbon and concentration
Bubble, Radix Isatidis after must soaking；
（2）Radix Isatidis takes dried object to be placed in air-flow crushing, crosses 100 mesh sieves, collect sieving in 40 ~ 50 DEG C of dryings after taking immersion
Grain, takes sieving particle to be placed in 55 ~ 60 DEG C of constant weights, takes constant weight sieving particle Steam explosion treatment, Radix Isatidis after must handling；
（3）According to the mass fraction, 7 ~ 10 parts of rhamnolipid is taken, 20 ~ 25 parts of cellulase, 5 ~ 8 parts of phenylalanine, polyethylene glycol 5 ~
8 parts, mass fraction is 70% 60 ~ 90 parts of ethanol solution, and 1000 parts of mixing of water, obtain soak, Radix Isatidis adds leaching after taking processing
Soaked in bubble liquid, take out Radix Isatidis, it is dry, obtain the pretreatment Radix Isatidis for the extraction Banlangen Polysaccharide that ferments.
2. the Radix Isatidis preprocess method according to claim 1 for being used for fermentation extraction Banlangen Polysaccharide, it is characterised in that
The step（1）Middle plant culture：According to the mass fraction, 60 ~ 70 parts of ammonium nitrate, 100 ~ 110 parts of potassium nitrate, biphosphate are taken
120 ~ 130 parts of potassium, 200 ~ 250 parts of magnesium sulfate, 140 ~ 150 parts of calcium chloride, 6-BA0.02 ~ 0.06 part, NAA0.02 ~ 0.05 part, sugarcane
20 ~ 30 parts of sugar, 20 ~ 25 parts of agar, 400 ~ 500 parts of water, obtain plant culture.
3. the Radix Isatidis preprocess method according to claim 1 for being used for fermentation extraction Banlangen Polysaccharide, it is characterised in that
The step（1）Middle Radix Isatidis explant, which is placed in the ethanol solution that mass fraction is 70%, soaks 30 ~ 40s, is placed in mass fraction
To soak 5 ~ 6min in 0.1% mercuric chloride solution；Illumination cultivation condition：23 ~ 27 DEG C of cultures 5 ~ 7d, intensity of illumination 2000lx,
Daily light application time is 12h；Cultivate the quality of back plate indigo plant root explant and activated carbon and concentration for 0.01mol/L citric acid solutions
Than for 10：1：0.3 ~ 0.5, soak 2 ~ 2.5h.
4. the Radix Isatidis preprocess method according to claim 1 for being used for fermentation extraction Banlangen Polysaccharide, it is characterised in that
The step（2）Middle steam blasting condition is：150MPa, pressurize 150s.
5. the Radix Isatidis preprocess method according to claim 1 for being used for fermentation extraction Banlangen Polysaccharide, it is characterised in that
The step（3）The mass ratio of Radix Isatidis and soak is 1 after middle processing：5~7；30 ~ 35 DEG C of 30 ~ 40min of immersion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711246490.3A CN107987178A (en) | 2017-12-01 | 2017-12-01 | A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711246490.3A CN107987178A (en) | 2017-12-01 | 2017-12-01 | A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107987178A true CN107987178A (en) | 2018-05-04 |
Family
ID=62034991
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711246490.3A Pending CN107987178A (en) | 2017-12-01 | 2017-12-01 | A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107987178A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109608558A (en) * | 2019-01-10 | 2019-04-12 | 山东农业大学 | A kind of extracting method of natural plant polyose |
| CN111304266A (en) * | 2019-12-12 | 2020-06-19 | 武汉新华扬生物股份有限公司 | Biological enzymolysis fermentation process for aloe |
| CN114790338A (en) * | 2022-04-26 | 2022-07-26 | 安徽和美生态农业科技有限公司 | Preparation method and application of antibacterial edible composite film |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1994331A (en) * | 2006-12-30 | 2007-07-11 | 中国人民解放军第三军医大学第一附属医院 | Super micronization method for botanical medicinal, product obtained thereby and use thereof |
-
2017
- 2017-12-01 CN CN201711246490.3A patent/CN107987178A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1994331A (en) * | 2006-12-30 | 2007-07-11 | 中国人民解放军第三军医大学第一附属医院 | Super micronization method for botanical medicinal, product obtained thereby and use thereof |
Non-Patent Citations (6)
| Title |
|---|
| COSTAS G. BILIADERIS等著，王元凤等翻译: "《功能性食品碳水化合物》", 30 April 2009, 中国轻工业出版社 * |
| JIHONG LI等: ""the mechanism of poly(ethylene glycol) 4000 effect on enzymatic hydrolysis of lignocellulose"", 《COLLOIDS AND SURFACES B:BIOINTEFACES》 * |
| 张晓阳等: "《纤维素生物质水解与应用》", 31 December 2012, 郑州大学出版社 * |
| 曹墨菊主编: "《植物生物技术概论》", 31 October 2014, 中国农业大学出版社 * |
| 梅家训等: "《组培快繁技术及其应用》", 30 April 2003, 中国农业出版社 * |
| 陆强等: "《液体生物燃料技术与工程》", 31 January 2013, 上海科学技术出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109608558A (en) * | 2019-01-10 | 2019-04-12 | 山东农业大学 | A kind of extracting method of natural plant polyose |
| CN111304266A (en) * | 2019-12-12 | 2020-06-19 | 武汉新华扬生物股份有限公司 | Biological enzymolysis fermentation process for aloe |
| CN114790338A (en) * | 2022-04-26 | 2022-07-26 | 安徽和美生态农业科技有限公司 | Preparation method and application of antibacterial edible composite film |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101596220B (en) | Wall-breaking method of ganoderma lucidum spore | |
| CN111317694B (en) | Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics | |
| CN104187746B (en) | The technique that fermentation method prepares Testa Phaseoli radiati water soluble dietary fiber | |
| CN106581083A (en) | Extraction method for ganoderma lucidum components, biological feed and preparation method thereof | |
| CN106491662B (en) | A method of polysaccharide and flavones are extracted simultaneously from Phellinus cultivation waste material | |
| CN108823261A (en) | A kind of Ultra-low molecular weight Dendrobium officinale polysaccharide and its preparation and application | |
| CN107987178A (en) | A kind of Radix Isatidis preprocess method for being used for fermentation extraction Banlangen Polysaccharide | |
| CN103689550A (en) | Ganoderma lucidum functional food and preparation method thereof | |
| CN106916861A (en) | It is a kind of while the method for producing Auricularia polysaccharide and melanin | |
| CN102488087A (en) | Biological detoxification method for camellia seed cakes | |
| CN107893029B (en) | Composite wall-breaking treatment method of ganoderma lucidum spore powder | |
| CN112869179A (en) | Process for extracting dietary fiber from sweet potatoes | |
| CN109576324A (en) | A kind of astragalus polyose and its biological extraction method | |
| CN104357332B (en) | Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid | |
| CN112608949B (en) | Preparation method and application of pseudo-ginseng flower extract | |
| CN104844583B (en) | A kind of production method of Puerarin | |
| CN114085778A (en) | Rhizopus oryzae JYH-4-23 and application thereof in extraction of chlorogenic acid in honeysuckle | |
| CN110628846B (en) | Method for preparing xylooligosaccharide by high-temperature high-pressure treatment | |
| CN112773752A (en) | Whitening anti-aging raw material and preparation method and application thereof | |
| CN102533565B (en) | Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed | |
| US20230148629A1 (en) | Traditional chinese medicine polysaccharide extract freeze-drying protective agent, direct-to-vat starter and preparation methods thereof | |
| CN108796027A (en) | A method of producing carotenoid | |
| CN115820783A (en) | Preparation method for obtaining multiple triterpenoids by fermenting rosa roxburghii tratt | |
| CN106947796A (en) | A kind of D trehaloses purifying technique | |
| CN109206475B (en) | Method for simultaneously extracting oil body and protein by single-step enzymolysis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180504 |
|
| RJ01 | Rejection of invention patent application after publication |