CN104013657A - Post-fermentation extracting method of extracting saponin from American ginseng medicinal material - Google Patents
Post-fermentation extracting method of extracting saponin from American ginseng medicinal material Download PDFInfo
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Abstract
The invention relates to a microbial fermentation re-extracting method after extracting saponin from an American ginseng medicinal material. The method comprises the following steps: (1) drying and smashing the American ginseng medicinal material with saponin extracted, filling in a fermentation tank, adding an aqueous solution dissolved with ammonium sulfate and dipotassium phosphate, and uniformly mixing; (2) preparation of a zymophyte seed solution, namely carrying out activated and multiplication culture on Aspergillus niger and bacillus subtilis to obtain a mixed zymophyte seed solution; (3) inoculating the mixed zymophyte seed solution to the American ginseng medicinal material, mixing and pre-treating, fermenting for a fixed time under a stable condition, terminating fermentation and drying to obtain the fermentation extract of the American ginseng medicinal material. The American ginseng fermentation extract provided by the invention is rich in low peptide of proteins and American ginseng polysaccharide components, high in nutritional value and good in taste, and the extraction yield is remarkably higher than that of the conventional traditional Chinese medicine water-decocting extracting method.
Description
Technical field
The present invention relates to medical material fermented extracted field, refer to especially a kind of to carrying out microorganism fermentation extracting method again after American ginseng medicine saponin extraction.
Background technology
Radix Panacis Quinquefolii (Panax quinquefolium L.) is araliaceae ginseng plant, originates in northern US to the southern band of Canada, and China started to introduce a fine variety in the seventies in last century.The conventional underground rhizome of Radix Panacis Quinquefolii is used as medicine, and is the Chinese herbal medicine that China's Ministry of Public Health approval can be used for health food, has boosting qi and nourishing yin, and effect of clearing away heat and promoting production of body fluid is lost for the deficiency of vital energy is cloudy, interior-heat, and cough with asthma expectorant blood, deficiency-heat is tired tired, quenches one's thirst, dryness of the mouth and throat.
Radix Panacis Quinquefolii is rich in saponin, polysaccharide, starch, aminoacid, protein, crude fibre, trace element etc., wherein saponin component is considered to the most effective active substance in Radix Panacis Quinquefolii, thereby pharmacy corporation concentrates on the extraction of saponin constituent to the extraction of Radix Panacis Quinquefolii, conventionally adopt ethanol percolate extraction, the Radix Panacis Quinquefolii tailing that is rich in the water-soluble substanceses such as polysaccharide, protein, aminoacid after extraction as garbage processed fall.Modern study confirmation, Radix Panacis Quinquefolii polysaccharide, protide composition have good biological activity equally, and Radix Panacis Quinquefolii polysaccharide has reducing effect to the blood glucose extremely increasing, and can reduce the infringement of radioactive material confrontation body, promotes the synthetic of body nucleic acid and protein.Radix Panacis Quinquefolii albumen has equally the body's immunity, resisting fatigue, blood fat reducing, the anti-good fortune of enhancing of adjusting and penetrates and analgesic activity.Thereby the comprehensive utilization of American ginseng medicine is studied, improve product resource utilization rate, expand the product using value of Radix Panacis Quinquefolii, will there is wide market prospect, bring considerable economic benefit.
At present, fermentation process is applied in Chinese medicine extraction, has become important research direction and the approach of comprehensive utilization exploitation natural resources of Chinese medicinal materials.Chinese medicine is taking plant medical material as main, its effective ingredient is present in the endochylema of cell more, and the cell wall of plant cell is by cellulose, hemicelluloses etc. form, its compact structure, if thereby adopt direct instructions of taking, most functional components are wherein difficult to be absorbed by the body, bioavailability is low, adopt solvent extraction as decocted, the methods such as percolation, can only extract certain constituents, easily cause the loss of other functional components or nutritional labeling, abandon, adopt biofermentation method, microbial inoculant is fermented in target Chinese medicine, the cellulase that utilizes microorganism to produce in metabolic process, the multiple exoenzyme such as pectase enters culture medium, thereby cause Chinese crude drug cell rupture, its effective ingredient and nutritional labeling are released, the protease that macromolecular substances also can be produced by fermentation process as plant proteins simultaneously etc. is degraded, form the small-molecule active substance that human body easily absorbs.Compared with Chinese medicine processing mode, modern Chinese medicine fermentation process has high superiority.
Summary of the invention
The invention provides a kind of method of American ginseng medicine being carried out to microorganism fermented extracted, American ginseng medicine extracts after saponin through the present invention's extract obtained protein content that ferments can reach 25.1%, polyoses content reaches 3.76%, deduct blank microorganism fermentation gained, extract yield and can improve respectively 37%, 50% compared with Chinese medicine water boiling and extraction method, crude fiber content reduces by 56% simultaneously, makes products therefrom nutrition more balanced.The low peptide protein that in gained fermented product extract of the present invention, molecular weight is less than 6500Da accounts for 85% of total protein, and more conventional extraction method doubles, and products therefrom nutritive value is higher.
Fermentation technique is the traditional food-processing method of China, along with the development of modern biological method, fermentation technique is applied in the industries such as agricultural, food, medicine, petrochemical industry and the energy widely, microbial fermentation processes is applied in Chinese medicine extraction, being the Innovative Development of Chinese medicine processing, is the high-tech herbal pharmaceutical new method forming in conjunction with modern microbial project.
This patent is selected aspergillus niger and bacillus subtilis to carry out American ginseng medicine and is extracted saponin after fermentation extraction, aspergillus niger is common food fermentation bacterial strain, be widely used as production food additive, comprise production enzyme preparation and organic acid etc., U.S. food drug surveilance office (FDA) is defined as safe food additive amyloglucosidase, catalase, pectase, oxidase and the lipase in aspergillus niger source, and China also safe fermentation strain using aspergillus niger as extensive use is used so far.The abundant enzymes such as aspergillus niger can secreting amylase, cellulase, pectase, protease, phytase, these enzymes can impel the macromolecular compound such as starch, cellulose in raw material to be degraded to nutrient substance, need for growth of microorganism; Bacillus subtilis can produce strong active protease, and some high molecular weight protein of can degrading generates little peptide and the aminoacid with high nutritive value, produces and is used widely in the oligopeptide class of Semen sojae atricolor at present.The present invention passes through process for solid state fermentation, aspergillus niger and bacillus subtilis are carried out to combination and compatibility, extract tailing by fermentation American ginseng medicine, obtain and preferably produce enzyme and efficient degradation effect, not only significantly improve the extraction yield of polysaccharide, protein-based nutritive effect composition, and the low peptide content in extraction product is increased than also, has improved product nutritive value.
Extract American ginseng medicine by this patent, reaction condition gentleness, the protein, the polysaccharide component that improve in product extract yield, increase the content ratio of low peptide simultaneously, and its leaching process comprises the following steps:
(1) American ginseng medicine extracting after saponin discharging is dried, is pulverized, and packs in fermentation tank, adds that to be dissolved with the aqueous solution of ammonium sulfate, potassium hydrogen phosphate appropriate, mixes.
American ginseng medicine powder is 1:0.5-1:1.5 with the mass ratio that adds water, and preferably mass ratio is 1:0.75.
American ginseng medicine powder is 1:0.002:0.0025-1:0.08:0.015 with the mass ratio that adds ammonium sulfate, dipotassium hydrogen phosphate, and preferably mass ratio is 1:0.04:0.005.
(2) preparation of zymocyte seed liquid: the activation of aspergillus niger and bacillus subtilis, amplification cultivation, obtain mixed culture fermentation bacterium seed liquor.
Wherein the activation method of aspergillus niger is that picking one encircles from original strain inclined-plane, and method of scoring is inoculated on culture medium A inclined-plane, is placed in 28 DEG C of incubators, cultivates 3-4 days, obtains aspergillus strain seed.
The activation method of bacillus subtilis is that picking one encircles from original strain inclined-plane, and method of scoring is inoculated on culture medium B inclined-plane, is placed in 30 DEG C of incubators, cultivates 3-4 days, obtains bacillus subtilis thalline seed.
The amplification method of aspergillus niger, bacillus subtilis is that aspergillus strain seed is inoculated in the triangular flask that culture medium A is housed by the inoculum concentration of culture medium weight 3-6%, and 29 DEG C of constant temperature, 180rpm shaking table are cultivated 24-36h, obtain aspergillus niger seed.Equally bacillus subtilis thalline seed is inoculated in the triangular flask that culture medium B is housed by the inoculum concentration of culture medium weight 3-6%, 29 DEG C of constant temperature, 180rpm shaking table, cultivate 24-36h, obtains bacillus subtilis seed.Aspergillus niger seed is mixed in 3:1-1:3 ratio with bacillus subtilis seed, and preferred proportion 1:2, obtains mixed culture fermentation bacterium seed liquor.
The prescription of culture medium A feature is: potato ball 80%, and glucose 10%, agar 10%, pH value is 7,121 DEG C of sterilizing 20min.
Culture medium B feature prescription is beerwort 99.8%, agar 0.2%, pH nature, 121 DEG C of sterilizing 20min.
(3) American ginseng medicine pretreatment fluid inoculation mixed culture fermentation bacterium seed liquor, fermentation, obtains fermentation liquid.
Wherein American ginseng medicine pretreatment fluid inoculation method is, by mixed culture fermentation bacterium seed liquor by volume the inoculum concentration of 4-8% under aseptic condition, be inoculated in fermentation tank.
Fermentation process is controlled fermentation temperature 35-37 DEG C, and fermentation 4-6 days, stops fermentation, dries, and obtains Western Radix Panacis Quinquefolii tailing fermented product extract.
Gained zymocyte extract of the present invention can be by adding adjuvant, then through filling or canned or pelletizing press sheet, make the acceptable preparation such as capsule or powder or tablet.
Adjuvant is excipient, binding agent, disintegrating agent, lubricant, flavoring agent etc., and excipient is selected from one or more the combination in starch, mannitol, sorbitol, lactose, glucose, cellulose, microcrystalline Cellulose, calcium phosphate, calcium carbonate.Preferably microcrystalline cellulose and lactose.Binding agent is selected from one or more the combination in cellulose, methylcellulose, hydroxypropyl cellulose, hydroxypropyl emthylcellulose, hydroxy methocel, polyvidone, gelatin, arabic gum, Polyethylene Glycol, sucrose, starch.Preferably polyvidone and hydroxypropyl cellulose.Disintegrating agent is selected from one or more the combination in the hydroxypropyl cellulose, sodium bicarbonate, sodium phosphate, sodium citrate of starch, hydroxy methocel, polyvinylpolypyrrolidone, low degree of substitution.The preferably hydroxypropyl cellulose of polyvinylpolypyrrolidone and low degree of substitution.Lubricant is selected from one or more the combination in sodium stearate, magnesium stearate, light anhydrous silicic acid, Pulvis Talci and sodium lauryl sulphate.Preferably magnesium stearate.Flavoring agent is selected from one or more the combination in citric acid, menthol, glycerol, orange-seed oil.Optimization citric acid.
Brief description of the drawings
Fig. 1 is solid-state fermentation process route.
Detailed description of the invention
Embodiment mono-: Radix Panacis Quinquefolii extracts the optimization of fermentation condition
1. polysaccharide component assay in tunning:
Adopt sulfuric acid-phynol method, measure with reference to the assay method of water solublity crude polysaccharides in health food.
1. solid-state fermentation process route
As shown in Figure 1.
2 fermented bacteriums are selected
The solid fermentation experiment of 2.1 single cultures
Now selecting five kinds of food conventional fermentation mycete and bacillus cereus to carry out single culture Radix Panacis Quinquefolii tailing fermenting experiment investigates.Select root aspergillus oryzae, aspergillus niger, rhizopus, Bacillus licheniformis, bacillus subtilis, beer yeast and seven kinds of fermented bacteriums of Candida utilis, respectively Radix Panacis Quinquefolii medicine tailing is carried out to the experiment of single culture solid fermentation, investigate the impact of different strain on the rear polyoses content of Radix Panacis Quinquefolii medicine tailing fermentation.Five kinds of strains are inoculated in 20g culture medium by 5% inoculum concentration respectively, add one times of dry medical material moisture of amount, at 29 DEG C of temperature, ammonium sulfate addition 2% condition bottom fermentation 5 days, measure polyoses content in the rear butt of fermentation.The results are shown in Table 1:
The experiment of table 1 single culture solid fermentation
As can be seen from the above table, in mycete, in aspergillus niger, bacillus cereus, bacillus subtilis gained polyoses content is maximum, thereby selects these two kinds of bacterium to carry out mixed culture fermentation Radix Panacis Quinquefolii tailing.
The solid fermentation experiment of the different proportionings of 2.2 mixed bacterias
Aspergillus niger after cultivating is mixed according to 3:1,2:1,1:1,1:2,1:3 ratio with bacillus subtilis, be seeded in 20g culture medium by 5% inoculum concentration, add one times of water gaging of Radix Panacis Quinquefolii tailing, ammonium sulfate addition 2%, mixes, 29 DEG C of fermentation temperatures, pH nature, ferment 5 days, polyoses content in the 5th day sampling and measuring culture medium, the results are shown in Table 2:
The experiment of table 2 mixed bacteria solid fermentation
From upper table can with find out, in the time that aspergillus niger and bacillus subtilis ratio are 1:2, the crude polysaccharides content of fermentation finished product reaches the highest.If aspergillus niger is too much, tunning musty is larger, affects products taste.If access aspergillus niger is very few, saccharifying enzyme and the cellulase of secretion are less, are unfavorable for cellulosic decomposition in Radix Panacis Quinquefolii, thereby to select aspergillus niger and bacillus subtilis ratio be 1:2.
3. affect the single-factor experiment of fermentation condition
Taking polyoses content as index, investigate respectively inorganic nitrogen-sourced selection and addition, raw material particle size, solid-to-liquid ratio, original ph, fermentation time, single bacterium and mixed fermentation Experimental Comparison and mixed vaccine proportioning determine, mixed fermentation condition.
The investigation of 3.1 nitrogenous source additions
Chinese medicine mostly is herbaceous plant, and nitrogenous source lacks relatively, adds in right amount nitrogenous source and is conducive to Microbial puotein production and breeding.In solid medium, add water by one times of amount of Radix Panacis Quinquefolii tailing, select ammonium sulfate as nitrogenous source, add nitrogenous source in the ratio of the dry medical material of 0-8%, mixed bacteria (aspergillus niger and bacillus subtilis, in the proportioning ratio of 1:2, are added to 5% inoculum concentration) is seeded in 20g culture medium to 29 DEG C of fermentation temperatures, pH nature, ferment 5 days, polyoses content in the 5th day sampling and measuring culture medium, the results are shown in Table 3:
Table 3 nitrogenous source addition is investigated
Can be found out by table, nitrogenous source addition is too much or very few, all unfavorable to the extraction of crude polysaccharides: in the time that nitrogenous source addition is 4%, in tunning, crude polysaccharides content reaches the highest.During lower than this addition because the not enough thalli growth breeding of nitrogenous source is slow; During higher than this addition, the osmotic pressure of culture medium is too high causes adverse influence to microbial growth.
3.2K
2hPO
4addition is investigated
Potassium ion is one of cation important in biological cell, is the activator of many enzymes, can promote the metabolism of carbohydrate, and the important substance of the synthetic thalline nucleic acid backbone of phosphate radical, can promote the breeding of microorganism simultaneously, thereby to K
2hPO
4addition is investigated, and adds one times of water gaging in solid medium, adds ammonium sulfate by 4% Radix Panacis Quinquefolii tailing weight, and adds K in the ratio of the dry medical material of 0-1.5%
2hPO
4, mixed bacteria (aspergillus niger and bacillus subtilis, in the proportioning ratio of 1:2, are added to 5% inoculum concentration) is seeded in 20g culture medium, 29 DEG C of fermentation temperatures, pH nature, ferments 5 days, polyoses content in the 5th day sampling and measuring culture medium, the results are shown in Table 4:
Table 4K
2hPO
4addition is investigated
Can draw K from experimental result
2hPO
4addition while being greater than 0.25%, in tunning, crude polysaccharides content reaches high value, preferred addition is 0.5%.
3.3 fermentation times are investigated
In solid medium by adding 1 times of amount to be dissolved with the aqueous solution of the potassium hydrogen phosphate of ammonium sulfate, the 0.5% Radix Panacis Quinquefolii tailing weight of 4% Radix Panacis Quinquefolii tailing weight, by mixed bacteria (by aspergillus niger and bacillus subtilis in the proportioning ratio of 1:2, add 5% inoculum concentration) be seeded in 20g culture medium, 29 DEG C of fermentation temperatures, pH nature, ferment 7 days, polyoses content in the 2nd, 3,4,5,6 days difference sampling and measuring culture medium of fermentation, the results are shown in Table 5:
Table 5 fermentation time is investigated
Microorganism fermentation is along with the consumption of the nutrient substance in culture medium, micro organism quantity becomes logarithm level to increase, according to upper table result, while fermenting by the 4th day, polyoses content reaches peak value substantially, and along with the increase of time, in culture medium, nutrient substance, oxygen and space reduce gradually, microbial reproduction speed eases up gradually, and it is slow that the increase of microbial metabolic products is also tending towards thereupon.Because fermentation time is long, easily make culture medium be polluted and other strains of growing, simultaneously in order to reduce costs and energy consumption, select 4-6 days the bests of incubation time, be preferably 5 days.
3.4 adequate moisture Content inspects
For solid fermentation, the initial moisture of culture medium is a key factor that affects microorganism fungus kind Growth and reproduction, by mixed bacteria (by aspergillus niger and bacillus subtilis in the proportioning ratio of 1:2, add 5% inoculum concentration) be seeded in 20g culture medium, add respectively 25%, 50%, 75%, 100%, 150% moisture, add ammonium sulfate by 4% Radix Panacis Quinquefolii tailing weight, add K by 0.5% Radix Panacis Quinquefolii tailing weight
2hPO
4, 29 DEG C of fermentation temperatures, pH nature, ferments 5 days, and polyoses content in the 5th day sampling and measuring culture medium of fermentation, the results are shown in Table 6:
The impact of table 6 moisture on solid fermentation
According to upper table result, water content is in the time of 50%-100%, and polyoses content is maximum, while continuing to increase, polyoses content declines to some extent, this may be because Aspergillus niger is aerobic organism, and too much moisture causes the aeration of culture medium to decline, and is unfavorable for the growth of Aspergillus niger and produces enzyme.Thereby to select moisture be 50%-100%, preferably 75%.
Embodiment bis-: Radix Panacis Quinquefolii extracts saponin after fermentation and extracts
The activation method of aspergillus niger is that picking one encircles from original strain inclined-plane, and method of scoring is inoculated on culture medium A inclined-plane, is placed in 28 DEG C of incubators, cultivates 3-4 days, obtains aspergillus strain seed.
The activation method of bacillus subtilis is that picking one encircles from original strain inclined-plane, and method of scoring is inoculated on culture medium B inclined-plane, is placed in 30 DEG C of incubators, cultivates 3-4 days, obtains bacillus subtilis thalline seed.
The amplification method of aspergillus niger, bacillus subtilis is that aspergillus strain seed is inoculated in the triangular flask that culture medium is housed by the inoculum concentration of culture medium weight 3-6%, and 29 DEG C of constant temperature, 180rpm shaking table are cultivated 24h, obtain aspergillus niger seed.Equally bacillus subtilis thalline seed is inoculated into by the inoculum concentration of culture medium weight 5% in the triangular flask that culture medium is housed, 29 DEG C of constant temperature, 180rpm shaking table, cultivate 24h, obtains bacillus subtilis seed.Aspergillus niger seed is mixed in 1:2 ratio with bacillus subtilis seed, obtain mixed culture fermentation bacterium seed liquor.
Get Radix Panacis Quinquefolii and extract tailing oven dry, adopt pulverizer 30 orders of pulverizing and sieve, getting crushing rear material 5kg packs in fermentation tank, 0.2kg ammonium sulfate, 0.025kg dipotassium hydrogen phosphate are dissolved in 3.75L water, and add in fermentation tank, mix homogeneously, by mixed culture fermentation bacterium seed liquor by volume 6% inoculum concentration under aseptic condition, be inoculated in fermentation tank, 38 DEG C of controlled fermentation temperature, ferment 5 days, stop fermentation, dry, obtain Radix Panacis Quinquefolii and extract saponin after fermentation extract (Fermentation-Assisted Extraction Samples, FAES).
Embodiment tri-: Radix Panacis Quinquefolii Different Extraction Method contrast
By conventional extraction method extract (Traditional Extraction Samples after gained Radix Panacis Quinquefolii extraction saponin after fermentation extract (FAES) in example two and Radix Panacis Quinquefolii extraction saponin, TES) and with glucose substitute American ginseng medicine gained blank extract (the Control Samples that ferments, CS) contrast, three kinds of extract polysaccharide, protein, content of cellulose difference are contrasted, and FAES and two kinds of extracts of TES are further purified to protein, the variation of contrast range of molecular weight distributions.
The Radix Panacis Quinquefolii of getting drying and crushing extracts tailing dries, adopt pulverizer 30 orders of pulverizing and sieve, get crushing rear material 1kg, the 5kg that adds water directly carries out water-bath reflux, extract, 2 hours, gained extracting liquid filtering, concentrated, evaporated under reduced pressure, as the conventional extract of medical material (TES) after Radix Panacis Quinquefolii extraction saponin.
Separately substitute Radix Panacis Quinquefolii tailing 5kg with 0.5kg glucose and make an addition in the solid fermentation Mixed culture substrate of embodiment bis-, and microbe inoculation is with embodiment bis-fermented extracted operations, extract obtained as blank sample (CS).
To conventional extraction method extract (Traditional Extraction Samples after Radix Panacis Quinquefolii extraction saponin after fermentation extract (FAES) and Radix Panacis Quinquefolii extraction saponin, and blank extract (Control Samples TES), CS) protein, reducing sugar and crude fibre composition in are analyzed, protein content determination adopts Kjeldahl's method, with reference to the mensuration of GB5009.5-2010 Protein in Food; Content of reducing sugar is measured and is adopted 3,5-dinitrosalicylic acid method, with reference to the mensuration of reducing sugar in GB/T5009.7-2008 food; Crude fibre is measured and is adopted acid-alkali washing method to measure, with reference to the mensuration of dietary fiber in GB/T5009.88-2008 food.The results are shown in Table 7:
The component analysis of three kinds of extraction product nutrients of table 7
Upper table shows, in tunning, protein content reaches 25.10% with this understanding, the albumen that deduction blank extract produces, extract yield and increase by 37%, crude fiber content is 11.27%, and degradation rate is 56.60%, and the content of crude polysaccharides is 3.74, except the crude polysaccharides that blank extract produces, extracting yield increases approximately 50%.
The mensuration that polypeptide molecular weight distributes:
Get respectively 50mmol/L Tris-HCl (pH8.3) buffer that conventional extraction method extract (TES) and fermented extracted method extract (FAES) add 5 times of volumes in right amount, be slowly ground to evenly; At the centrifugal 30min of rotating speed of 5000r/min, get supernatant, in precipitation, add again 50mmol/L Tris-HCl (pH8.3) buffer of 5 times of amounts to carry out secondary lixiviate, centrifugal, supernatant merges; Add solid (NH4) 2SO4 to final concentration be 80%, stir, hold over night at 4 DEG C, with the centrifugal 30min of 5000r/min, distilled water dissolves for precipitation, and water is fully dialysed, lyophilizing, obtains American ginseng total albumen.
Adopt gel filtration chromatography molecular weight distribution.Chromatographic condition: chromatographic column is TSKgelG2000SWXL300mm × 7.8mm gel column; Mobile phase is acetonitrile: water: trifluoroacetic acid=45:55:0.1; Flow velocity is 0.5mL/min; Sampling volume is 50 μ L; Detection wavelength is 220nm.
Molecular weight mark product are bovine serum albumin (molecular weight MW67000Da), cytochrome (MW12500Da), aprotinin (MW6500Da), cobamide (VBl2, MW1355.38Da), glutathion (MW307.32Da).
Sample solution preparation: by American ginseng total protein sample 100mg, put in 10ml volumetric flask, the phosphate buffered solution dilution that adds 10mM pH7.4 is settled to scale.
Measurement result sees the following form:
According to upper table result, the protein that Radix Panacis Quinquefolii fermented product extract is purified, molecular weight is less than the ratio of 6500Da apparently higher than conventional extraction method gained, illustrate and adopt fermentation method to extract Radix Panacis Quinquefolii tailing, can not only improve the extraction yield of protein, holoside nutrient material, can also increase low molecular weight peptide class material proportion, optimize trophic structure, improve nutritive value.
Claims (8)
1. to carrying out a method for microorganism fermented extracted after American ginseng medicine extraction saponin, it is characterized in that extracting by following steps:
(1) American ginseng medicine extracting after saponin is dried, pulverized, pack in fermentation tank, add that to be dissolved with the aqueous solution of ammonium sulfate, dipotassium hydrogen phosphate appropriate, mix;
(2) preparation of zymocyte seed liquid: select aspergillus niger and bacillus subtilis is activated, amplification cultivation, obtain mixed culture fermentation bacterium seed liquor;
(3) American ginseng medicine powder inoculation mixed culture fermentation bacterium seed liquor, mixes after pretreatment, in the steady-state conditions bottom fermentation set time, stops fermentation, dries, and obtains American ginseng medicine fermented product extract.
2. a kind of American ginseng medicine according to claim 1 extracts saponin after fermentation extracting method, it is characterized in that: in step (1), American ginseng medicine powder is 1:0.5-1:1.5 with the mass ratio that adds water.
3. a kind of American ginseng medicine according to claim 1 extracts saponin after fermentation extracting method, it is characterized in that: in step (1), American ginseng medicine powder is 1:0.002:0.0025-1:0.08:0.015 with the mass ratio that adds ammonium sulfate, dipotassium hydrogen phosphate.
4. a kind of American ginseng medicine according to claim 1 extracts saponin after fermentation extracting method, it is characterized in that: in step (2), zymocyte selects one or more bacterium in aspergillus oryzae, aspergillus niger, rhizopus, Bacillus licheniformis, bacillus subtilis, beer yeast, Candida utilis to cultivate, obtain single or mixed culture fermentation bacterium seed liquor, preferably aspergillus niger and bacillus subtilis carry out respectively seed culture.
5. a kind of American ginseng medicine according to claim 4 extracts saponin after fermentation extracting method, it is characterized in that: in step (2), the preferred aspergillus niger of zymocyte carries out respectively mixing after seed culture with bacillus subtilis, obtains mixed culture fermentation bacterium seed liquor.
6. a kind of American ginseng medicine extracts saponin after fermentation extracting method according to claim 1 or 5, it is characterized in that: in step (2), select zymocyte aspergillus niger, bacillus subtilis activation, the amplification cultivation of warp respectively, obtain aspergillus niger seed and bacillus subtilis seed, aspergillus niger seed is mixed in 3:1-1:3 ratio with bacillus subtilis seed, obtain mixed culture fermentation bacterium seed liquor.
7. a kind of American ginseng medicine according to claim 1 extracts saponin after fermentation extracting method, it is characterized in that: after Radix Panacis Quinquefolii medicinal material extract saponin, pretreatment fluid inoculation method is in step (3), by mixed culture fermentation bacterium seed liquor by volume the inoculum concentration of 4-8% under aseptic condition, be inoculated in fermentation tank.
8. extract saponin after fermentation extracting method according to a kind of American ginseng medicine described in claim 1 or 7, it is characterized in that: in step (3), fermentation process is controlled fermentation temperature 35-37 DEG C, fermentation 4-6 days, stops fermentation, dry, obtain American ginseng medicine fermented product extract.
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