CN104013657B - A kind of American ginseng medicine extracts after saponin(e microbial fermentation extracting method again - Google Patents
A kind of American ginseng medicine extracts after saponin(e microbial fermentation extracting method again Download PDFInfo
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Abstract
American ginseng medicine is extracted the present invention relates to one kind microbial fermentation extracting method again is carried out after saponin(e, comprised the following steps：（1）The American ginseng medicine drying after saponin(e will be extracted, crushed, be fitted into fermentation tank, addition is appropriate dissolved with the aqueous solution of ammonium sulfate, dipotassium hydrogen phosphate, mixes；（2）The preparation of zymophyte seed liquor：Aspergillus niger and bacillus subtilis are activated, amplification cultivation, obtain mixed fermentation bacterium seed liquor；（3）American ginseng medicine powder is inoculated with mixed fermentation bacterium seed liquor, after mixing pretreatment, is fermented the set time under stable condition, terminates fermentation, drying, produces American ginseng medicine extractive from fermentative.Gained American Ginseng extractive from fermentative of the invention is rich in the low peptide of albumen and Radix Panacis Quinquefolii polysaccharide constituents, is of high nutritive value, in good taste, extracts yield and is significantly higher than Chinese medicine traditional water decoction and boils extraction method.
Description
Technical field
The present invention relates to medicinal material fermentation extraction field, micro- life is carried out after particularly relating to a kind of saponin extraction to American ginseng medicine
Thing fermentation extracting method again.
Background technology
American Ginseng（Panax quinquefolium L.）For araliaceae ginseng plant, originate in northern US to plus take
A big southern band, China starts to introduce a fine variety in the seventies in last century.American Ginseng is often used as medicine with underground rhizome, is the approval of the Ministry of Public Health of China
Available for the Chinese herbal medicine of health food, there is a boosting qi and nourishing yin, effect of clearing heat and promoting fluid, for deficiency of vital energy yin deficiency, interior heat, cough and asthma phlegm blood,
Abnormal heat is tired of tired, quenches one's thirst, dryness of the mouth and throat.
American Ginseng rich in saponin(e, polysaccharide, starch, amino acid, protein, crude fibre, trace element etc., wherein saponins into
It is considered as maximally effective active material in American Ginseng to divide, thus extraction of the pharmacy corporation to American Ginseng concentrates on saponin constituent
Extract, generally extracted using ethanol percolation, the American Ginseng tail rich in water-soluble substanceses such as polysaccharide, protein, amino acid after extraction
Material is then disposed off as discarded object.Modern study confirms that Radix Panacis Quinquefolii polysaccharide, proteinaceous components equally have good biology living
Property, Radix Panacis Quinquefolii polysaccharide has reduction effect to the abnormal blood glucose increased, and can reduce the infringement of radioactive material confrontation body, promotees
Enter the synthesis of body nucleic acid and protein.American Ginseng albumen equally has regulation body's immunity, antifatigue, reducing blood lipid, increasing
Qiang Kangfu is penetrated and analgesic activity.Thus the comprehensive utilization to American ginseng medicine is studied, and improves product resource utilization rate, is expanded
The products application value of American Ginseng, will have wide market prospects, bring considerable economic benefit.
At present, fermentation process is applied in traditional Chinese medicine extraction, it has also become the important research of comprehensive utilization exploitation natural resources of Chinese medicinal materials
Direction and approach.Chinese medicine is present in the endochylema of cell more based on plant medicinal material its active ingredient, and plant cell is thin
Cell wall is made up of cellulose, hemicellulose etc., its compact structure, thus if using direct instructions of taking, it is therein many
Number functional components are difficult to be absorbed by the body, and bioavilability is low, is such as decocted using solvent extraction, diacolation method, then can only carry
Certain constituents is taken, loss, the discarding of other functional components or nutritional ingredient are easily caused, using biofermentation method, by microorganism
It is inoculated in target Chinese medicine to be fermented, cellulase, pectase for producing in metabolic process using microorganism etc. are a variety of extracellular
Enzyme enters culture medium, so as to cause Chinese medicine cell rupture, its active ingredient and nutritional ingredient are released, while macromolecular complex
Protease that matter such as plant proteinses can also be produced by fermentation process etc. is degraded, and it is small that formation human body easily absorbs
Molecular activity material.Compared with traditional Chinese medicine processing mode, modern Chinese herbal medicine fermentation process has high superiority.
The content of the invention
The invention provides a kind of method that microbial fermentation extraction is carried out to American ginseng medicine, American ginseng medicine extracts soap
Extract obtained protein content is fermented up to 25.1% through the present invention after glycosides, and polyoses content deducts blank microorganism hair up to 3.76%
Obtained by ferment, 37%, 50% can be respectively increased compared with traditional Chinese medicine water boiling and extraction method by extracting yield, while crude fiber content reduction by 56%,
Make products therefrom nutrition more balanced.The low peptide albumen that gained extractive from fermentative middle-molecular-weihydroxyethyl of the invention is less than 6500Da accounts for total protein
85%, more conventional extraction method is doubled, and products therefrom nutritive value is higher.
Fermentation technique is the traditional food-processing method of China, and with the development of modern biotechnology method, fermentation technique is wide
General is applied in the industries such as agricultural, food, medicine, petrochemical industry and the energy, and microbial fermentation processes are carried applied to Chinese medicine
It is the Innovative Development of Chinese medicine preparation in taking, is the high-tech herbal pharmaceutical new method with reference to formed by Modern microbiological engineering.
This patent carries out American ginseng medicine from aspergillus niger and bacillus subtilis and extracts saponin(e after fermentation extraction, aspergillus niger
It is common food fermentation bacterial strain, is widely used as producing food additives, including production enzyme preparation and organic acid etc., U.S.'s food
Product drug surveilance office (FDA) is the amyloglucosidase of Aspergillus niger origin, catalase, pectase, oxidizing ferment and fat
Fat enzyme is defined as safe food additives, and China also uses aspergillus niger as wide variety of safe fermentation strain so far.
The enzyme that aspergillus niger can be enriched with secreting amylase, cellulase, pectase, protease, phytase etc., these enzymes can promote
The high-molecular compound such as starch, cellulose is degraded to nutriment in raw material, and being grown for microorganism needs；Bacillus subtilis
More strongly active protease can be produced, small peptide and amino of some high molecular weight protein generations that can degrade with high nutritive value
Acid, is used widely in the oligomeric peptides production of soybean at present.The present invention by process for solid state fermentation, by aspergillus niger and
Bacillus subtilis is combined compatibility, and by fermenting, American ginseng medicine extracts tailing, obtains preferably producing enzyme and efficient degradation
Effect, not only significantly improves polysaccharide, the extraction yield of protein-based nutritive effect composition, and extracts the low peptide in product and contain
Amount is increased than also, improves product nutritive value.
American ginseng medicine is extracted by this patent, reaction condition is gentle, improve the protein in product, polysaccharide component and extract
Yield, while increasing the content ratio of low peptide, its extraction process comprises the following steps：
（1）Extract the American ginseng medicine drying after saponin(e discharging, crush, be fitted into fermentation tank, add dissolved with ammonium sulfate, phosphorus
The aqueous solution of potassium hydrogen phthalate is appropriate, mixes.
The mass ratio of American ginseng medicine powder and addition water is 1：0.5-1:1.5, preferably mass ratio is 1：0.75.
The mass ratio of American ginseng medicine powder and addition ammonium sulfate, dipotassium hydrogen phosphate is 1:0.002:0.0025-1:0.08:
0.015, preferably mass ratio is 1：0.04：0.005.
（2）The preparation of zymophyte seed liquor：The activation of aspergillus niger and bacillus subtilis, amplification cultivation, obtain mixing hair
Yeast-like fungi seed liquor.
Wherein the activation method of aspergillus niger is that it is oblique that method of scoring is inoculated in culture medium A from the ring of picking one on original strain inclined-plane
On face, it is placed in 28 DEG C of incubators, cultivates 3-4 days, obtain aspergillus strain seed.
The activation method of bacillus subtilis is that method of scoring is inoculated in culture medium B from the ring of picking one on original strain inclined-plane
On inclined-plane, it is placed in 30 DEG C of incubators, cultivates 3-4 days, obtain bacillus subtilis thalline seed.
Aspergillus niger, the amplification method of bacillus subtilis are the inoculation that aspergillus strain seed is pressed to culture medium weight 3-6%
Amount is inoculated into the triangular flask equipped with culture medium A, 29 DEG C of constant temperature, 180rpm shaking tables, is cultivated 24-36h, is obtained aspergillus niger seed.
Equally bacillus subtilis thalline seed is inoculated into the triangular flask equipped with culture medium B by culture medium weight 3-6% inoculum concentration,
29 DEG C of constant temperature, 180rpm shaking tables, cultivate 24-36h, obtain bacillus subtilis seed.By aspergillus niger seed and bacillus subtilis
Strain presses 3:1-1:3 ratios are mixed, preferred proportion 1:2, obtain mixed fermentation bacterium seed liquor.
The prescription of culture medium A feature is：Potato ball 80%, glucose 10%, agar 10%, pH value is 7,121 DEG C of sterilizings
20min。
Culture medium B features prescription is brewer's wort 99.8%, agar 0.2%, pH naturally, 121 DEG C of sterilizing 20min.
（3）American ginseng medicine pretreatment fluid is inoculated with mixed fermentation bacterium seed liquor, and fermentation obtains zymotic fluid.
Wherein American ginseng medicine pretreatment fluid inoculation method is, by mixed fermentation bacterium seed liquor 4-8% by volume inoculation
Amount is aseptically inoculated into fermentation tank.
Fermentation process is 35-37 DEG C of fermentation temperature of control, is fermented 4-6 days, terminates fermentation, and drying obtains Western American Ginseng tail
Expect extractive from fermentative.
Gained fermentation fungus extract of the invention can be by adding auxiliary material, then by filling or canned or pelletizing press sheet, is made
The acceptable preparation such as capsule or pulvis or tablet.
Auxiliary material is excipient, adhesive, disintegrant, lubricant, flavor enhancement etc., and excipient is selected from starch, mannitol, mountain
Combination more than one or both of pears alcohol, lactose, glucose, cellulose, microcrystalline cellulose, calcium phosphate, calcium carbonate.It is preferred that
Microcrystalline cellulose and lactose.Adhesive is selected from cellulose, methylcellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxyl first
Combination more than one or both of base cellulose, PVP, gelatin, Arabic gum, polyethylene glycol, sucrose, starch.It is preferred that
PVP and hydroxypropyl cellulose.Disintegrant is selected from starch, hydroxymethyl cellulose, PVPP, the hydroxypropyl of low degree of substitution
Combination more than one or both of cellulose, sodium acid carbonate, sodium phosphate, sodium citrate.It is preferred that PVPP and low taking
Dai Du hydroxypropyl cellulose.Lubricant is selected from odium stearate, magnesium stearate, light anhydrous silicic acid, talcum powder and dodecyl
Combination more than one or both of sodium sulphate.It is preferred that magnesium stearate.Flavor enhancement is selected from citric acid, menthol, glycerine, orange
Combination more than one or both of oil.Optimization citric acid.
Brief description of the drawings
Fig. 1 is solid-state fermentation process route.
Embodiment
Embodiment one：American Ginseng extracts the optimization of fermentation condition
1. polysaccharide component assay in tunning：
Using sulfuric acid-phynol method, it is measured with reference to the assay method of Water-soluble polysccharide in health food.
1. solid-state fermentation process route
As shown in Figure 1.
2 fermented bacteriums are selected
The solid state fermentation experiment of 2.1 single cultures
Now select the conventional fermentation mould of five kinds of food to carry out single culture American Ginseng tailing fermenting experiment with bacillus to examine
Examine.Select root aspergillus oryzae, aspergillus niger, head mold, bacillus licheniformis, bacillus subtilis, brewer's yeast and candida utili
Seven kinds of fermented bacteriums, carry out single culture solid state fermentation experiment to American Ginseng medicine tailing respectively, investigate different strain to American Ginseng
The influence of polyoses content after the fermentation of medicine tailing.Five kinds of strains are inoculated into 20g culture mediums by 5% inoculum concentration respectively, one times is added
Dry medicinal material moisture is measured, is fermented 5 days under the conditions of 29 DEG C of temperature, ammonium sulfate addition 2%, polyoses content in butt after measure fermentation.
It the results are shown in Table 1：
The single culture solid state fermentation of table 1 is tested
As can be seen from the above table, polyoses content is most obtained by bacillus subtilis in aspergillus niger, bacillus in mould, because
And select both bacterium to carry out mixed fermentation American Ginseng tailing.
The solid state fermentation experiment of 2.2 mixed bacteria different ratios
By the aspergillus niger after cultivation and bacillus subtilis according to 3:1、2:1、1：1、1:2、1:3 ratios are mixed, and are pressed
5% inoculum concentration is seeded in 20g culture mediums, adds one times of amount water of American Ginseng tailing, and ammonium sulfate addition 2% is mixed, fermentation temperature
29 DEG C, pH are naturally, ferment 5 days, and polyoses content in the 5th day sampling and measuring culture medium the results are shown in Table 2：
The mixed bacteria solid state fermentation of table 2 is tested
Can be with finding out, when aspergillus niger and bacillus subtilis ratio are 1 from upper table:When 2, the Thick many candies content of fermented product
Reach highest.If aspergillus niger is excessive, tunning musty is larger, influences products taste.If it is very few to access aspergillus niger, secrete
Carbohydrase and cellulase it is less, be unfavorable for the decomposition of cellulose in American Ginseng, thus selection aspergillus niger and bacillus subtilis
Bacterium ratio is 1:2.
3. influence the single factor experiment of fermentation condition
Using polyoses content as index, inorganic nitrogen-sourced selection and addition, raw material particle size, solid-to-liquid ratio, initial pH are investigated respectively
Value, fermentation time, single bacterium and mixed fermentation Experimental Comparison and Mixed Microbes proportioning are determined, mixed fermentation condition.
The investigation of 3.1 nitrogen source additions
Chinese medicine is generally herbaceous plant, and nitrogen source is relative to be lacked, and appropriate addition nitrogen source is conducive to Microbial puotein production and breeding.
Added water in solid medium by one times of amount of American Ginseng tailing, selection ammonium sulfate is added as nitrogen source in the ratio of the dry medicinal materials of 0-8%
Plus nitrogen source, aspergillus niger and bacillus subtilis (are pressed 1 by mixed bacteria:2 ratio, adds 5% inoculum concentration) it is seeded to
In 20g culture mediums, 29 DEG C of fermentation temperature, pH are naturally, ferment 5 days, and as a result polyoses content in the 5th day sampling and measuring culture medium is shown in
Table 3：
The nitrogen source addition of table 3 is investigated
Nitrogen source addition is excessive or very few it can be seen from table, and the extraction to Thick many candies is all unfavorable：When nitrogen source addition is
When 4%, Thick many candies content reaches highest in tunning.Because the not enough thalli growth breeding of nitrogen source is slow during less than this addition；
During higher than this addition, the too high growth to microorganism of osmotic pressure of culture medium is adversely affected.
3.2K2HPO4Addition is investigated
Potassium ion is one of important cation in biological cell, is the activator of many enzymes, can promote carbohydrate
Metabolism, while the important substance of the synthesis thalline nucleic acid backbone of phosphate radical, can promote the breeding of microorganism, thus to K2HPO4
Addition is investigated, and one times of amount water is added in solid medium, ammonium sulfate is added by 4% American Ginseng tailing weight, and by 0-
The ratio addition K of 1.5% dry medicinal material2HPO4, aspergillus niger and bacillus subtilis (are pressed 1 by mixed bacteria:2 ratio, plus
Enter 5% inoculum concentration) be seeded in 20g culture mediums, 29 DEG C of fermentation temperature, pH naturally, fermentation 5 days, the 5th day be measured by sampling culture medium
Middle polyoses content, the results are shown in Table 4：
Table 4K2HPO4Addition is investigated
K can be drawn from experimental result2HPO4Addition when being more than 0.25%, Thick many candies content reaches height in tunning
Value, preferably addition are 0.5%.
3.3 fermentation times are investigated
Ammonium sulfate, 0.5% American Ginseng tailing in solid medium by 1 times of amount of addition dissolved with 4% American Ginseng tailing weight
The aqueous solution of the potassium hydrogen phosphate of weight, (1 is pressed by mixed bacteria by aspergillus niger and bacillus subtilis:2 ratio, is added
5% inoculum concentration) be seeded in 20g culture mediums, 29 DEG C of fermentation temperature, pH naturally, fermentation 7 days, in fermentation the 2nd, 3,4,5,6 days
Polyoses content in separately sampled measure culture medium, the results are shown in Table 5：
The fermentation time of table 5 is investigated
Microbial fermentation is with the consumption of the nutriment in culture medium, and micro organism quantity increases into Logarithmic degree, according to upper
Table result, when fermenting by the 4th day, polyoses content basically reaches peak value, increases over time, nutriment, oxygen in culture medium
Gas and space are gradually decreased, and microbial reproduction speed gradually eases up, and the increase of microbial metabolic products also tends to be slow therewith
Slowly.Because fermentation time is long, easily it is contaminated culture medium and grows other strains, while in order to reduce cost and energy
Consumption, selects incubation time 4-6 days most preferably, preferably 5 days.
3.4 adequate moisture contents are investigated
For solid fermentation, the initial moisture of culture medium is to influence a key factor of microorganism fungus kind growth and breeding,
Aspergillus niger and bacillus subtilis (are being pressed 1 by mixed bacteria:2 ratio, adds 5% inoculum concentration) it is seeded to 20g cultures
In base, 25%, 50%, 75%, 100%, 150% moisture is added respectively, ammonium sulfate is added by 4% American Ginseng tailing weight, by 0.5%
American Ginseng tailing weight adds K2HPO4, 29 DEG C of fermentation temperature, pH are naturally, ferment 5 days, and culture is measured by sampling in the 5th day in fermentation
Polyoses content in base, the results are shown in Table 6：
Influence of the moisture of table 6 to solid fermentation
According to upper table result, water content in 50%-100%, polyoses content for maximum, continue increase when polyoses content
Decline, this is probably that excessive moisture causes the aeration of culture medium to decline, is unfavorable for black song because black-koji mould is aerobic organism
The growth of mould and producing enzyme.Thus selection moisture is 50%-100%, preferably 75%.
Embodiment two：American Ginseng extracts saponin(e after fermentation and extracted
The activation method of aspergillus niger is that method of scoring is inoculated on culture medium A inclined-plane from the ring of picking one on original strain inclined-plane,
It is placed in 28 DEG C of incubators, cultivates 3-4 days, obtain aspergillus strain seed.
The activation method of bacillus subtilis is that method of scoring is inoculated in culture medium B from the ring of picking one on original strain inclined-plane
On inclined-plane, it is placed in 30 DEG C of incubators, cultivates 3-4 days, obtain bacillus subtilis thalline seed.
Aspergillus niger, the amplification method of bacillus subtilis are the inoculation that aspergillus strain seed is pressed to culture medium weight 3-6%
Amount is inoculated into the triangular flask equipped with culture medium, 29 DEG C of constant temperature, 180rpm shaking tables, is cultivated 24h, is obtained aspergillus niger seed.Equally
Bacillus subtilis thalline seed is inoculated into the triangular flask equipped with culture medium by the inoculum concentration of culture medium weight 5%, 29 DEG C of perseverances
Temperature, 180rpm shaking tables, cultivate 24h, obtain bacillus subtilis seed.Aspergillus niger seed and bacillus subtilis seed are pressed 1:
2 ratios are mixed, and obtain mixed fermentation bacterium seed liquor.
Take American Ginseng to extract tailing drying, crushed using pulverizer and 30 mesh that sieve, take crushing rear material 5kg to load hair
In fermentation tank, 0.2kg ammonium sulfate, 0.025kg dipotassium hydrogen phosphates are dissolved in 3.75L water, and added in fermentation tank, be well mixed,
By mixed fermentation bacterium seed liquor by volume 6% inoculum concentration be aseptically inoculated into fermentation tank, control fermentation temperature 38
DEG C, ferment 5 days, terminate fermentation, drying obtains American Ginseng and extracts saponin(e after fermentation extract (Fermentation-Assisted
Extraction Samples,FAES)。
Embodiment three：American Ginseng Different Extraction Method is contrasted
Gained American Ginseng in example two is extracted into saponin(e after fermentation extract (FAES) and routine after American Ginseng extraction saponin(e
Extraction method extract (Traditional Extraction Samples, TES) and with glucose substitute American ginseng medicine sent out
Blank control extract (Control Samples, CS) is contrasted obtained by ferment, by three kinds of extract polysaccharide, protein, fibers
Cellulose content difference is contrasted, and protein is further purified to FAES and two kinds of extracts of TES, contrasts range of molecular weight distributions
Change.
Take the American Ginseng of drying and crushing to extract tailing drying, crushed using pulverizer and 30 mesh that sieve, take crushing rear material
1kg, the 5kg that adds water directly carries out water-bath refluxing extraction 2 hours, the filtering of gained extract solution, concentration, evaporated under reduced pressure, is used as American Ginseng
Extract medicinal material conventional extract after saponin(e（TES）.
It is another that the solid fermentation mixed culture matrix that American Ginseng tailing 5kg makes an addition to embodiment two is substituted with 0.5kg glucose
In, and operation is extracted in the fermentation of microbe inoculation be the same as Example two, it is extract obtained to be used as blank control sample（CS）.
Saponin(e after fermentation extract (FAES) is extracted to American Ginseng and extracts traditional extraction method extract after saponin(e with American Ginseng
In (Traditional Extraction Samples, TES) and blank control extract (Control Samples, CS)
Protein, reduced sugar and coarse fiber content are analyzed, and protein content determination uses Kjeldahl's method, with reference to GB5009.5-
The measure of 2010 Protein in Food；Content of reducing sugar, which is determined, uses 3,5- dinitrosalicylic acid systems, with reference to GB/T5009.7-
The measure of reduced sugar in 2008 food；Crude fibre is determined to be determined using acid-alkali washing method, with reference to GB/T5009.88-2008 food
The measure of middle dietary fiber.It the results are shown in Table 7：
7 three kinds of extraction product nutrient constituent analyses of table
Upper table shows that protein content reaches 25.10% in tunning on this condition, deducts blank control and extracts produce
Raw albumen, extracts yield increase by 37%, crude fiber content is 11.27%, and degradation rate is 56.60%, and the content of Thick many candies is
3.74, except the Thick many candies that blank control extract is produced, extract yield increase about 50%.
The measure of polypeptide molecular weight distribution：
Traditional extraction method extract is taken respectively（TES）And fermentation extraction method extract（FAES）Appropriate 5 times of volumes of addition
50mmol/L Tris-HCl (pH8.3) buffer solution, is slowly ground to uniform；30min is centrifuged in 5000r/min rotating speed, is taken
Clear liquid, 50mmol/L Tris-HCl (pH8.3) buffer solution that 5 times of amounts are added into precipitation carries out secondary extraction, centrifuges, on
Clear liquid merges；Add solid (NH4) 2SO4 to final concentration of 80%, stir, stood overnight at 4 DEG C, with 5000r/min from
Heart 30min, precipitation distilled water is dissolved, and water is fully dialysed, and is freezed, is obtained American ginseng total albumen.
Using gel filtration chromatography molecular weight distribution.Chromatographic condition：Chromatographic column is TSKgelG2000SWXL300m
M × 7.8mm gel columns；Mobile phase is acetonitrile：Water：Trifluoroacetic acid=45:55:0.1；Flow velocity is 0.5mL/min；Sampling volume is
50μL；Detection wavelength is 220nm.
Molecular weight mark product are bovine serum albumin(BSA) (molecular weight MW67000Da), cytochromes (MW12500Da), Aprotinin
(MW6500Da), cobamide (VBl2, MW1355.38Da), glutathione (MW307.32Da).
It is prepared by sample solution：American ginseng total protein sample 100mg is put in 10ml volumetric flasks, plus 10mM pH7.4 phosphorus
The dilution of hydrochlorate cushioning liquid is settled to scale.
Measurement result see the table below：
According to upper table result, the protein that American Ginseng extractive from fermentative is purified, molecular weight is less than 6500Da ratio
Obtained by example apparently higher than traditional extraction method, illustrate to extract American Ginseng tailing using fermentation method, protein, polysaccharide battalion can not only be improved
Support the extraction yield of material, moreover it is possible to increase low molecular weight peptide class material proportion, optimize trophic structure, improve nutritive value.
Claims (7)
1. a kind of extract the method that microbial fermentation extraction is carried out after saponin(e to American ginseng medicine, it is characterised in that passes through following step
Suddenly extracted：
(1) the American ginseng medicine drying after saponin(e will be extracted, crushed, be fitted into fermentation tank, add dissolved with ammonium sulfate, phosphoric acid hydrogen two
The aqueous solution of potassium is appropriate, mixes；
(2) preparation of zymophyte seed liquor：Select that aspergillus niger and bacillus subtilis be activated, amplification cultivation, obtain mixing hair
Yeast-like fungi seed liquor；
(3) American ginseng medicine powder inoculation mixed fermentation bacterium seed liquor, after mixing pretreatment, when fermenting fixed under stable condition
Between, fermentation, drying are terminated, American ginseng medicine extractive from fermentative is produced.
2. a kind of American ginseng medicine according to claim 1 extracts saponin(e after fermentation extracting method, it is characterised in that：
The mass ratio of American Ginseng medicinal material powder and addition water is 1 in step (1)：0.5-1:1.5.
3. a kind of American ginseng medicine according to claim 1 extracts saponin(e after fermentation extracting method, it is characterised in that：
The mass ratio of American Ginseng medicinal material powder and addition ammonium sulfate, dipotassium hydrogen phosphate is 1 in step (1):0.002:0.0025-1:
0.08:0.015。
4. a kind of American ginseng medicine according to claim 1 extracts saponin(e after fermentation extracting method, it is characterised in that：
The preferred aspergillus niger of zymophyte is mixed after carrying out seed culture respectively with bacillus subtilis in step (2), is mixed
Zymophyte seed liquor.
5. a kind of American ginseng medicine according to claim 1 or 4 extracts saponin(e after fermentation extracting method, it is characterised in that：
Selection zymophyte aspergillus niger, bacillus subtilis pass through respectively in step (2) activation, amplification cultivation, obtain aspergillus niger seed and
Bacillus subtilis seed, 3 are pressed by aspergillus niger seed and bacillus subtilis seed:1-1:3 ratios are mixed, and are mixed
Zymophyte seed liquor.
6. a kind of American ginseng medicine according to claim 1 extracts saponin(e after fermentation extracting method, it is characterised in that：
Pretreatment fluid inoculation method is after American Ginseng medicinal material extract saponin(e in step (3), and mixed fermentation bacterium seed liquor is pressed into volume
Inoculum concentration than 4-8% is aseptically inoculated into fermentation tank.
7. a kind of American ginseng medicine according to claim 1 or 6 extracts saponin(e after fermentation extracting method, it is characterised in that：
Fermentation process is 35-37 DEG C of fermentation temperature of control in step (3), is fermented 4-6 days, terminates fermentation, and drying obtains American ginseng medicine hair
Ferment extract.
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| CN104643062A (en) * | 2015-01-26 | 2015-05-27 | 邱芳萍 | American ginseng ferment |
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| CN111574607B (en) * | 2020-05-12 | 2023-09-05 | 武汉市华甜生物科技有限公司 | Method for extracting thaumatin from African arrowroot based on microbial fermentation |
| CN112772913A (en) * | 2021-01-25 | 2021-05-11 | 昆明生物制造研究院有限公司 | American ginseng rose enzyme and preparation method thereof |
| CN113072599A (en) * | 2021-03-10 | 2021-07-06 | 武汉市华甜生物科技有限公司 | Method for preparing natural hesperidin by biological fermentation and extraction |
| CN113350428A (en) * | 2021-06-16 | 2021-09-07 | 山东中医药大学 | Fermentation method of radix tetrastigme |
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