CN102676342A - Truffle wine and preparation method thereof - Google Patents
Truffle wine and preparation method thereof Download PDFInfo
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- CN102676342A CN102676342A CN2012101765020A CN201210176502A CN102676342A CN 102676342 A CN102676342 A CN 102676342A CN 2012101765020 A CN2012101765020 A CN 2012101765020A CN 201210176502 A CN201210176502 A CN 201210176502A CN 102676342 A CN102676342 A CN 102676342A
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of foods or medicine and provides a truffle wine and a preparation method thereof. The preparation method enables truffle mycelium to be improved by 69%, truffle extracellular polysaccharide yield is improved by 150%, and bacterial intracellular polysaccharide is improved by 57%. The preparation method is simple and easy to operate and suitable for industrial production.
Description
Technical field
The invention belongs to medical technical field, be specifically related to healthcare products that contain ferfas mycelium powder or ferfas hypha fluid and preparation method thereof.
Background technology
Ferfas (Truffle) is also claimed truffle, has unique fragrance, mouthfeel and nutritive value, the history of the existing more than one thousand years of human consumption ferfas.Ferfas is rich in kind of physiologically active ingredient surplus 17 seed amino acids, 8 kinds of VITAMINs, an amount of protein and androsterone, sterol, sphingolipid, lipid acid, amino acid and the trace element etc. 50.And essential nutrients such as containing human body self can not synthetic 8 seed amino acid, zinc, manganese, iron, calcium, phosphorus, selenium; Have pharmaceutical uses such as strengthening immunity, anti-ageing, beneficial stomach, clear god, hemostasis, treatment hemorrhoid; Has antitumour activity; Cancer cells there is certain restraining effect, can excites brain cell activity.
The health promoting wine fragrance of ferfas infusion is unique, mouthfeel is good, and green health and pharmacological action are obvious, have establishing-Yang, kidney tonifying, and reduction serum low-density LP, SUV, prevent atherosclerosis and remarkable efficacy such as antitumor at high density lipoprotein increasing.
Bibliographical information has the mouse of S-180 sarcoma, EAC sarcoma liquid continuously through administration in 10 days inoculation, find the fast granulose ability obvious suppression mouse S-180 sarcoma of 25mg/kg, 50mg/kg and the growth of EAC sarcoma.Find that through successive administration truffle polysaccharides has tangible increase effect to the weight of mouse spleen, leukocyte count and T lymphocyte percentage in the peripheral blood simultaneously; Can promote the T lymphocyte transformation, improve in the mice serum IgG level etc.Experimental result shows, truffle polysaccharides is as a kind of new fungus polysaccharide, and toxicity is low, good water solubility, function of tumor inhibition are obvious.
Truffle polysaccharides can directly extract from kames, also can obtain through the method for ferfas liquid submerged fermentation, but the former is inappropriate for industrialized production, and reason is:
1, ferfas grows in subterranean medicine-food two-purpose fungi as a kind of, requires the environment that grows in alkaline soil, have a moderate climate at occurring in nature, and with the root system symbiosis of seeds such as Oak Tree, harsh growth conditions has directly caused natural output, and supply falls short of demand simultaneously;
2, for remedying the deficiency of the natural egg laying amount of ferfas, once there were many scholars to carry out the research that half manual simulation cultivates, but generally needed the 7-9 year from being inoculated into results, the cycle is longer, needs the man power and material of labor;
3, because ferfas grows in undergroundly, and the artificial difficulty of finding is bigger, is vulnerable to damage in the gatherer process, ferfas grows in nature and receives the influence of environmental change bigger simultaneously, and quality cannot say for sure card;
4, the fresh ferfas of selling in the market costs an arm and a leg, therefore if directly then can't avoid the restriction that receives cost of material and quantity with kames as the raw materials for production of truffle polysaccharides.
At present; Chinese patent 200610166503, one Chinese patent application 200810172343 disclose the method for producing ferfas active mycelium body and truffle polysaccharides; All through slant culture, level liquid seed culture, secondary liquid seeds are cultivated and the process of liquid submerged fermentation obtains ferfas active mycelium body and truffle polysaccharides.
Summary of the invention
Through lot of test, we increase the content that solid state fermentation can be crossed effective raising truffle polysaccharides during the fermentation at surprised discovery.The product of fermentation process gained of the present invention is with the content of the vitriol oil-phynol method determination block granulose; Mensuration result shows; Method of the present invention is than existing batch culture; The black truffle mycelium has improved 20% than cultivate in batches in the cultured products, and especially truffles exopolysaccharide rate ratio batch culture method has improved 50%, and the ferfas intracellular polyse has increased by 24.8% than culture method in batches in the ferfas cell cultures.
Simultaneously; We also find in substratum, to add the hazel leaf juice of suitable weight ratio; Can obviously improve the mycelial output of ferfas; The black truffle mycelium has improved 69.2% than cultivate in batches in the cultured products, and especially truffles exopolysaccharide rate ratio batch culture method has improved 150%, and the ferfas intracellular polyse has increased by 57.3% than culture method in batches in the ferfas cell cultures.
The purpose of this invention is to provide a kind of ferfas wine and preparation method thereof.
The preparation method who the purpose of this invention is to provide a kind of ferfas mycelium powder or ferfas hypha fluid.
The purpose of this invention is to provide a kind of solid state fermentation that is suitable for cultivating ferfas.
The purpose of this invention is to provide a kind of prescription that is suitable for cultivating the nutritive water of ferfas.
Particularly, the invention provides:
A kind of ferfas wine, its raw material comprise the ferfas mycelium of 5-95 weight part, the cereal of 95-5 weight part.
Described ferfas is mycelial preparation method may further comprise the steps:
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh kames, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 20-35 ℃ of constant incubator and cultivates 2-30 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 15-20 minute mistake and was filtered filtrating; 100-300g hazel leaf is put into 1000ml water boiled to cross in about 10-15 minute and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 10-30g again, agar 10-30g is settled to 1000ml with zero(ppm) water, and adjust pH is 6-9;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 15-40 ℃; Rotating speed is on 50-300 rev/min the rotary or reciprocating type bottle swingging machine shaking culture 2-15 days, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 300-900, and sucrose 1-300, peptone 1-100, yeast extract paste 1-20, sal epsom 1-10, potassium primary phosphate 1-10, the pH value is 6-9;
3) the secondary liquid seeds is cultivated: with step 2) in the level liquid seed cultivated transfer according to the inoculum size of 5-50% volume ratio and shaking of secondary liquid seed culture medium is housed carries out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 15-40 ℃; Rotating speed is on 50-300 rev/min the rotary or reciprocating type bottle swingging machine shaking culture 2-15 days, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 300-900, and sucrose 1-300, peptone 1-100, yeast extract paste 1-20, sal epsom 1-10, potassium primary phosphate 1-10, the pH value is 6-9;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 10-50g, yeast extract paste 0-20g, peptone 0-20g, potassium primary phosphate 0-10g, sal epsom 0-10g is dissolved in the 700-900ml water, is settled to 1000ml, regulates pH to 6-9;
2) configuration solid-state fermentation culture medium: by weight for (0.8-2): 1 joins cereal in the nutritive water;
3) solid state fermentation: according to the 1-50% volume/weight of solid fermentation substratum dry weight; The secondary liquid seeds is inoculated in step 2) on the solid-state fermentation culture medium of gained; In 20-40 ℃ of fermentation culture 2-60 days; Bed thickness is 1-10cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
Described ferfas is mycelial preparation method also comprise:
III, liquid submerged fermentation method are produced ferfas mycelium powder or ferfas hypha fluid;
1) configuration liquid nutrient medium: the bright hazel leaf of 100-300g is put into 1000ml water boil 20-40 minute after-filtration, get filtrating, add the sucrose of 50-200g; The peptone of 10-100g, the yeast extract paste of 5-50g, the sal epsom of 1-40g; The potassium primary phosphate of 1-40g; Water is settled to 1000ml, and adjust pH is 6-9, and sterilization promptly gets;
2) fermentation condition is: according to the inoculum size of 5-50% volume ratio the secondary liquid seeds of gained among the I is inoculated in 1) on the liquid nutrient medium of gained; Liquid amount is volumetrical 60-80%; At 20-40 ℃; Ventilation Rate be the 30-60 liter/minute, mixing speed is to cultivate 2-30 days under 200-800 rev/min the condition, the ferfas hypha fluid;
3) with step 2) gained ferfas hypha fluid is centrifugal, filters, and drying is pulverized, and sieves, and sterilization promptly gets the ferfas mycelium powder.
A kind of preparation method of ferfas wine: with weight ratio is 1: cereal (0.1-2) and ferfas mycelium powder or ferfas hypha fluid are as raw material; After the koji of access 2-8% weight ratio and the activated yeast mixture of 1-5% weight ratio; At 20-40 ℃ of bottom fermentation 2-10 days; At 15-30 ℃ of standing for fermentation 20-50 days,, process the ferfas yellow rice wine then according to the making method of yellow rice wine.
A kind of preparation method of ferfas wine: the ferfas mycelium fermentation powder of claim 2 gained is used ultrasonic disruption, 120 ℃ boiling 1-3 hour, put to room temperature; Every 100g ferfas mycelium fermentation powder adds yeast-saccharifying enzyme-acetic acid premixed liquid of 50-150mL; 20-40 ℃ ferment at constant temperature 10-30 days, use filtered through gauze then, filtrate 4-8 ℃ the fermentation 20-80 days, get supernatant, the millipore filtration degerming, the sealing, low tempertaure storage promptly gets; It is the 3-8% Glacial acetic acid min. 99.5 that described yeast-saccharifying enzyme-acetic acid premixed liquid comprises volume(tric)fraction, yeast 5-15g/100mL, saccharifying enzyme 10-20g/100mL.
Described Tuber bacterial strain is selected from one or more in Chinese ferfas, India truffle, black truffle, white ferfas or the summer truffle.
Described cereal is rice, millet, corn, purple rice, red rice, black rice, one or more of wheat, barley, highland barley, buckwheat or Chinese sorghum.
Actication of culture is meant that the bacterial classification of preservation state is put into the suitable culture base to be cultivated, enlarged culturing step by step, obtain to flush, the enough cultures of inoculation quantity.Strain fermentation has generally needs the 2-3 rejuvenation process in generation because the condition when preserving often the condition when cultivating is inequality, so want activation, let bacterial classification adapt to culture environment gradually.
Described yellow rice wine is meant with grain to be the brewing wine of raw material, does not comprise the distillatory liquor.
The present invention compared with prior art has the following advantages and positively effect:
1, can obviously improve the mycelial output of ferfas, improve 69% than culture method in batches.
2, can obviously improve truffles exopolysaccharide output, improve 150% than culture method in batches;
3, can obviously improve the ferfas intracellular polyse, improve 57% than culture method in batches;
4, preparation method of the present invention is simple, is fit to industrial production.
Embodiment
Below description through embodiment the present invention is described further; But this is not to be limitation of the present invention; Those skilled in the art are according to basic thought of the present invention; Can make various modifications or improvement, but only otherwise break away from basic thought of the present invention, all within scope of the present invention.
China ferfas, India truffle, black truffle, white ferfas, summer truffle all converge precious garden ferfas ltd available from Kunming.
Test Example 1
Comparative Examples (batch culture method):
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh black truffle sporophore, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 25 ℃ of constant incubators and cultivates 10 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 17 minutes mistakes and was filtered filtrating; Add sucrose 15g again, agar 15g is settled to 1000ml with zero(ppm) water, and adjust pH is 7;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 20 ℃; Rotating speed is a shaking culture 5 days on 100 rev/mins the reciprocating type bottle swingging machine, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: sucrose 50, and peptone 10, yeast extract paste 5, sal epsom 3, potassium primary phosphate 7, the pH value is 7;
3) the secondary liquid seeds is cultivated: with step 2) in the level liquid seed cultivated transfer according to the inoculum size of 10% volume ratio and shaking of secondary liquid seed culture medium is housed carries out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 20 ℃; Rotating speed is a shaking culture 5 days on 150 rev/mins the rotary or reciprocating type bottle swingging machine, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: sucrose 100, and peptone 30, yeast extract paste 10, sal epsom 5, potassium primary phosphate 4, the pH value is 7;
II, liquid submerged fermentation method are produced the ferfas hypha fluid;
1) configuration liquid nutrient medium: get the sucrose of 100g, the peptone of 30g, the yeast extract paste of 15g, the sal epsom of 10g, the potassium primary phosphate of 5g is settled to 1000ml with zero(ppm) water, and adjust pH is 7, and sterilization promptly gets behind the packing test tube;
2) inoculation, cultivate: the ferfas mycelium fermentation powder of gained among the II is inoculated in 1 according to 10% weight ratio) on the liquid nutrient medium of gained; At 25 ℃; Ventilation Rate is 35 liters/minute, and rotating speed is a shaking culture 13 days on 250 rev/mins the reciprocating type bottle swingging machine, the ferfas hypha fluid.
2) fermentation condition is: according to the inoculum size of 10% volume ratio the secondary liquid seeds of gained among the I is inoculated in 1) on the liquid nutrient medium of gained; Liquid amount is a volumetrical 70%; At 30 ℃; Ventilation Rate is 50 liters/minute, and mixing speed is to cultivate 15 days under 600 rev/mins the condition, the ferfas hypha fluid.The truffle polysaccharides content detecting method: cell is weighed after 60 ℃ of oven dry, and the cell of oven dry is according to document (Tang, Y.J.; Zhu, L.L., Li.D.S.; Mi; Z.Y, Li, H.M..Significance ofinoculation density and carbon source on the mycelia growth and Tuberpolysaccharides production by submerged fermentation of Chinese truffle Tubersinense.Process Biochemistry 2008; 43:576-586.) the method break process after, with the content of the vitriol oil-phynol method determination block granulose.
Comparative Examples, embodiment 1, embodiment 3 are carried out following assay determination:
The mensuration of A, exopolysaccharides: under 30 μ m aperture membrane filtrations dried cell weight; Add the long-pending absolute ethyl alcohol of tetraploid in the fermented liquid, mixing is spent the night, and obtains truffle polysaccharides outside the born of the same parents in 13000rpm centrifugation.Through the 1mol/L dissolution of sodium hydroxide, adopt the vitriol oil-phynol method to measure exopolysaccharides.
The mensuration of B, intracellular polyse output: mycelium 100mg, add the 1mol/L dissolution of sodium hydroxide, adopt the vitriol oil-phynol method to measure intracellular polyse output, calculate intracellular polyse output (intracellular polyse content multiply by dried cell weight).
The different cultural methods of table 1 are to the influence of ferfas mycelium and polysaccharide yield
Compare * * p＜0.01 with control group, compare #p＜0.05 for 1 group with medicine
Can know from table 1; Compare with Comparative Examples, the active mycelium scale of construction of embodiment 1 and embodiment 3 has obtained to increase significantly, than Comparative Examples; Embodiment 1 has increased 20%; Embodiment 3 has increased 69.1%, and showing increases the solid state fermentation step and in substratum, add hazel leaf juice, can promote the production that rises of ferfas active mycelium body.
In addition, also can know from table 1 that the truffle polysaccharides among embodiment 1 and the embodiment 3 is compared opposition also significantly to be increased, and embodiment 3 increases obviously also than the truffle polysaccharides of embodiment 1, wherein exocellular polysaccharide increases by 66%, and intracellular polyse increases by 26%.
Test Example 2 improves immunity test
Experimental animal: BALB/C male mice, body weight 18-20g.
Trial drug:
1 group of medicine: the product that Comparative Examples obtains in the Test Example 1
2 groups of medicines: the product that embodiment 9 obtains
3 groups of medicines: the product that embodiment 10 obtains
TP: get 20 of mouse, random packet becomes 5 groups, is hypoimmunity with the mouse modeling, and not modeling of normal group, modeling successfully begin administration, and control group gives saline water 0.2ml/20g.Administration 1-3 organizes gastric infusion, dosage 30ml/kg, successive administration 30 days; 24 hours broken end sacrificed by exsanguination mouse after the drug withdrawal; Under aseptic condition, take out spleen, process splenocyte suspension, mouse boosting cell is adjusted to 5 * 10 with RPMI-1640 (containing 10% calf serum)
6Cell/ml is added in 96 well culture plates, every hole 100 μ l, and each hole adds double (promptly adds ConA, and does not add ConA).ConA 20 μ l (concentration is 1000 μ g/ml), RPMI-1640 80 μ l are every hole to 200 μ l volumes, in 5%CO
2Cultivated 72 hours in 37 ℃ of thermostat containers, stop cultivating preceding 24 hours, every hole adds 3H-TdR3.7 * 10
9(iucr).With cell harvestor (TITERTEK CELL HARVESTER 550) collecting cell on 49 type glass fiber filter paper; Use zero(ppm) water, 5% triamine acetic acid, absolute ethanol washing successively; Place 60 ℃ of incubators to dry filter paper, be added in the cup that contains the 5ml scintillation solution, survey through liquid scintillation instrument (BECKMEN LS 9800) and participate in activity (cpm); Calculate SI (SI) then, promptly SI=adds ConA hole cpm/ and does not add ConA hole cpm.
The influence that table 2 different pharmaceutical transforms the immunocompromised mouse spleen lymphocyte.
| Group | SI |
| Normal group | 0.689±0.071** |
| Control group | 0.125±0.125 |
| 1 group of medicine | 1.321±0.103** |
| 2 groups of medicines | 3.120±0.631**# |
| 3 groups of medicines | 3.096±0.583**# |
Compare * * p＜0.01 with control group, compare #p＜0.05 for 1 group with medicine
Can know that by table 2 the prepared ferfas wine of the present invention can make the immunocompromised mouse spleen lymphocyte transform significantly to be increased.
The experiment of Test Example 3 mouse anti-reflecting fatigues
Experimental animal: Kunming mouse, body weight 18-20g.
Trial drug:
1 group of medicine: the product that Comparative Examples obtains in the Test Example 1
2 groups of medicines: the product that embodiment 9 obtains
3 groups of medicines: the product that embodiment 10 obtains
TP: get 20 of mouse, random packet becomes 4 groups, and control group gives saline water 0.2ml/20g.Administration 1-3 organizes gastric infusion, dosage 30ml/kg, and administration is put into the swimming pool of getting ready in advance with mouse after 1 hour fast, and record mouse is gone into the pond and is begun to time of swimming no longer.The result sees table 3.
Table 3 anti-fatigue test result
| Group | Swimming time (min) |
| Control group | 2.5±0.8 |
| 1 group of medicine | 4.6±0.3** |
| 2 groups of medicines | 8.4±0.6**# |
| 3 groups of medicines | 8.6±0.8**# |
Compare * * p＜0.01 with control group, compare #p＜0.05 for 1 group with medicine
Can know by table 2, but the swimming time of the prepared ferfas wine significant prolongation mouse of the present invention explains that medicinal liquor of the present invention has fine antifatigue effect.
Embodiment 1 (preparation ferfas mycelium fermentation powder)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh black truffle sporophore, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 25 ℃ of constant incubators and cultivates 15 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 16 minutes mistakes and was filtered filtrating; 200g hazel leaf is put into 1000ml water boiled to cross in about 15 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 20g again, agar 20g is settled to 1000ml with zero(ppm) water, and adjust pH is 7;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 30 ℃; Rotating speed is a shaking culture 8 days on 200 rev/mins the rotary shaker, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 600, and sucrose 150, peptone 50, yeast extract paste 10, sal epsom 5, potassium primary phosphate 5, the pH value is 7;
3) the secondary liquid seeds is cultivated: with the step 2 of 75ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 25 ℃; Rotating speed is a shaking culture 8 days on 150 rev/mins the rotary shaker, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 500, and sucrose 100, peptone 45, yeast extract paste 8, sal epsom 6.5, potassium primary phosphate 4.7, the pH value is 7;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 30g, yeast extract paste 10g, peptone 15g, potassium primary phosphate 5g, sal epsom 5g is dissolved in the 800ml water, is settled to 1000ml, regulates pH to 7;
2) configuration solid-state fermentation culture medium: the 1500g Chinese sorghum is joined in the 1000g nutritive water;
3) solid state fermentation: 38ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 30 ℃ of fermentation culture 40 days, bed thickness was 5cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
Embodiment 2 (preparation ferfas mycelium fermentation powder)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh Chinese kames, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 33 ℃ of constant incubators and cultivates 25 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 17 minutes mistakes and was filtered filtrating; 150g hazel leaf is put into 1000ml water boiled to cross in about 12 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 15g again, agar 15g is settled to 1000ml with zero(ppm) water, and adjust pH is 7.5;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 20 ℃; Rotating speed is a shaking culture 5 days on 100 rev/mins the reciprocating type bottle swingging machine, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 400, and sucrose 10, peptone 90, yeast extract paste 5, sal epsom 3, potassium primary phosphate 6, the pH value is 7.5;
3) the secondary liquid seeds is cultivated: with the step 2 of 25ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 20 ℃; Rotating speed is a shaking culture 5 days on 100 rev/mins the reciprocating type bottle swingging machine, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 450, and sucrose 10, peptone 7, yeast extract paste 17, sal epsom 3.1, potassium primary phosphate 8.6, the pH value is 7.5;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 15g, yeast extract paste 5g, peptone 18g, potassium primary phosphate 3g, sal epsom 8g is dissolved in the 850ml water, is settled to 1000ml, regulates pH to 7.5;
2) configuration solid-state fermentation culture medium: the 1000g buckwheat is joined in the 1000g nutritive water;
3) solid state fermentation: 15ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 25 ℃ of fermentation culture 10 days, bed thickness was 3cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
Embodiment 3 (preparation ferfas mycelium powder)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh black truffle sporophore, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 25 ℃ of constant incubators and cultivates 15 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 16 minutes mistakes and was filtered filtrating; 200g hazel leaf is put into 1000ml water boiled to cross in about 15 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 20g again, agar 20g is settled to 1000ml with zero(ppm) water, and adjust pH is 7;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 30 ℃; Rotating speed is a shaking culture 8 days on 200 rev/mins the rotary shaker, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 600, and sucrose 150, peptone 50, yeast extract paste 10, sal epsom 5, potassium primary phosphate 5, the pH value is 7;
3) the secondary liquid seeds is cultivated: with the step 2 of 75ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 25 ℃; Rotating speed is a shaking culture 8 days on 150 rev/mins the rotary shaker, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 500, and sucrose 100, peptone 45, yeast extract paste 8, sal epsom 6.5, potassium primary phosphate 4.7, the pH value is 7;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 30g, yeast extract paste 10g, peptone 15g, potassium primary phosphate 5g, sal epsom 5g is dissolved in the 800ml water, is settled to 1000ml, regulates pH to 7;
2) configuration solid-state fermentation culture medium: the 1500g Chinese sorghum is joined in the 1000g nutritive water;
3) solid state fermentation: 38ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 30 ℃ of fermentation culture 40 days, bed thickness was 5cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
III, liquid submerged fermentation method are produced the ferfas mycelium powder
1) configuration liquid nutrient medium: the bright hazel leaf of 200g is put into 1000ml water boil 30 minutes after-filtration, get filtrating, add the sucrose of 150g, the peptone of 50g; The yeast extract paste of 25g, the sal epsom of 20g, the potassium primary phosphate of 25g; Water is settled to 1000ml, and adjust pH is 7, and sterilization promptly gets;
2) fermentation condition is: the secondary liquid seeds 5ml of gained among the I is inoculated in 1) on the liquid nutrient medium 20ml of gained; Liquid amount is a volumetrical 70%, and at 30 ℃, Ventilation Rate is 45 liters/minute; Mixing speed is to cultivate 15 days under 500 rev/mins the condition, the ferfas hypha fluid;
3) with step 2) gained ferfas hypha fluid is centrifugal, filters, and drying is pulverized, and sieves, and sterilization promptly gets the ferfas mycelium powder.
Embodiment 4 (preparation ferfas hypha fluid)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh white kames, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 28 ℃ of constant incubators and cultivates 20 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 18 minutes mistakes and was filtered filtrating; 250g hazel leaf is put into 1000ml water boiled to cross in about 13 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 25g again, agar 25g is settled to 1000ml with zero(ppm) water, and adjust pH is 8;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 25 ℃; Rotating speed is a shaking culture 10 days on 150 rev/mins the rotary or reciprocating type bottle swingging machine, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 700, and sucrose 100, peptone 10, yeast extract paste 15, sal epsom 7, potassium primary phosphate 2, the pH value is 8;
3) the secondary liquid seeds is cultivated: with the step 2 of 50ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 30 ℃; Rotating speed is a shaking culture 10 days on 200 rev/mins the rotary or reciprocating type bottle swingging machine, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 650, and sucrose 150, peptone 56, yeast extract paste 3, sal epsom 8.2, potassium primary phosphate 6.5, the pH value is 8;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 25g, yeast extract paste 15g, peptone 3g, potassium primary phosphate 8g, sal epsom 3g is dissolved in the 750ml water, is settled to 1000ml, regulates pH to 8;
2) configuration solid-state fermentation culture medium: the 1800g black rice is joined in the 1000g nutritive water;
3) solid state fermentation: 30ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 35 ℃ of fermentation culture 20 days, bed thickness was 8cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
III, liquid submerged fermentation method are produced the ferfas hypha fluid;
1) configuration liquid nutrient medium: the bright hazel leaf of 150g is put into 1000ml water boil 25 minutes after-filtration, get filtrating, add the sucrose of 100g, the peptone of 30g; The yeast extract paste of 15g, the sal epsom of 5g, the potassium primary phosphate of 30g; Water is settled to 1000ml, and adjust pH is 8, and sterilization promptly gets;
2) fermentation condition is: the secondary liquid seeds 2ml of gained among the I is inoculated in 1) on the liquid nutrient medium 20ml of gained; Liquid amount is a volumetrical 65%, and at 25 ℃, Ventilation Rate is 40 liters/minute; Mixing speed is to cultivate 5 days under 400 rev/mins the condition, the ferfas hypha fluid.
Embodiment 5 (preparation ferfas mycelium powder)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh summer truffle sporophore, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 30 ℃ of constant incubators and cultivates 10 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 19 minutes mistakes and was filtered filtrating; 280g hazel leaf is put into 1000ml water boiled to cross in about 14 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 13g again, agar 28g is settled to 1000ml with zero(ppm) water, and adjust pH is 8.5;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 35 ℃; Rotating speed is a shaking culture 13 days on 250 rev/mins the rotary or reciprocating type bottle swingging machine, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 850, and sucrose 250, peptone 6, yeast extract paste 17, sal epsom 8, potassium primary phosphate 1-10, the pH value is 8.5;
3) the secondary liquid seeds is cultivated: with the step 2 of 100ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 35 ℃; Rotating speed is a shaking culture 13 days on 250 rev/mins the rotary or reciprocating type bottle swingging machine, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 750, and sucrose 250, peptone 84, yeast extract paste 13, sal epsom 2.9, potassium primary phosphate 7.3, the pH value is 8.5;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 45g, yeast extract paste 18g, peptone 10g, potassium primary phosphate 6.4g, sal epsom 2.5g is dissolved in the 870ml water, is settled to 1000ml, regulates pH to 8.5;
2) configuration solid-state fermentation culture medium: the 1300g corn is joined in the 1000g nutritive water;
3) solid state fermentation: 60ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 28 ℃ of fermentation culture 50 days, bed thickness was 2cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
III, liquid submerged fermentation method are produced the ferfas mycelium powder;
1) configuration liquid nutrient medium: the bright hazel leaf of 250g is put into 1000ml water boil 35 minutes after-filtration, get filtrating, add the sucrose of 180g, the peptone of 80g; The yeast extract paste of 45g, the sal epsom of 30g, the potassium primary phosphate of 80g; Water is settled to 1000ml, and adjust pH is 8.5, and sterilization promptly gets;
2) fermentation condition is: the secondary liquid seeds 8ml of gained among the I is inoculated in 1) on the liquid nutrient medium 20ml of gained; Liquid amount is a volumetrical 75%, and at 35 ℃, Ventilation Rate is 50 liters/minute; Mixing speed is to cultivate 20 days under 600 rev/mins the condition, the ferfas hypha fluid;
3) with step 2) gained ferfas hypha fluid is centrifugal, filters, and drying is pulverized, and sieves, and sterilization promptly gets the ferfas mycelium powder.
Embodiment 6 (preparation ferfas mycelium powder)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh India truffle sporophore, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 35 ℃ of constant incubators and cultivates 2 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 15 minutes mistakes and was filtered filtrating; 100g hazel leaf is put into 1000ml water boiled to cross in about 15 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 10g again, agar 10g is settled to 1000ml with zero(ppm) water, and adjust pH is 6;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 15 ℃; Rotating speed is a shaking culture 15 days on 50 rev/mins the reciprocating type bottle swingging machine, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 300, and sucrose 1, peptone 1, yeast extract paste 1, sal epsom 1, potassium primary phosphate 1, the pH value is 6;
3) the secondary liquid seeds is cultivated: with the step 2 of 12.5ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 15 ℃; Rotating speed is a shaking culture 15 days on 50 rev/mins the reciprocating type bottle swingging machine, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 300, and sucrose 1, peptone 1, yeast extract paste 1, sal epsom 1, potassium primary phosphate 1, the pH value is 6;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 10g, potassium primary phosphate 10g, sal epsom 10g is dissolved in the 700ml water, is settled to 1000ml, regulates pH to 6;
2) configuration solid-state fermentation culture medium: the 800g highland barley is joined in the 1000g nutritive water;
3) solid state fermentation: 1.5ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 20 ℃ of fermentation culture 60 days, bed thickness was 10cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
III, liquid submerged fermentation method are produced the ferfas mycelium powder
1) configuration liquid nutrient medium: the bright hazel leaf of 300g is put into 1000ml water boil 20 minutes after-filtration, get filtrating, add the sucrose of 200g, the peptone of 100g; The yeast extract paste of 50g, the sal epsom of 40g, the potassium primary phosphate of 40g; Water is settled to 1000ml, and adjust pH is 6, and sterilization promptly gets;
2) fermentation condition is: the secondary liquid seeds 1ml of gained among the I is inoculated in 1) on the liquid nutrient medium 20ml of gained; Liquid amount is a volumetrical 80%, and at 20 ℃, Ventilation Rate is 60 liters/minute; Mixing speed is to cultivate 30 days under 800 rev/mins the condition, the ferfas hypha fluid;
3) with step 2) gained ferfas hypha fluid is centrifugal, filters, and drying is pulverized, and sieves, and sterilization promptly gets the ferfas mycelium powder.
Embodiment 7 (preparation ferfas hypha fluid)
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh black truffle and white kames; Remove surperficial silt; Meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 20 ℃ of constant incubators and cultivates 30 days, gets slant strains; Described slant culture based formulas is: with yam 200g peeling, section was boiled 20 minutes mistakes and was filtered filtrating; 300g hazel leaf is put into 1000ml water boiled to cross in about 10 minutes and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose 30g again, agar 30g is settled to 1000ml with zero(ppm) water, and adjust pH is 9;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 40 ℃; Rotating speed is a shaking culture 2 days on 300 rev/mins the rotary shaker, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium (g/L) is: hazel leaf juice 900, and sucrose 300, peptone 100, yeast extract paste 20, sal epsom 10, potassium primary phosphate 10, the pH value is 9;
3) the secondary liquid seeds is cultivated: with the step 2 of 125ml) in transfer 250ml that the secondary liquid seed culture medium is housed of the level liquid seed cultivated shake and carry out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 40 ℃; Rotating speed is a shaking culture 2 days on 300 rev/mins the rotary shaker, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium (g/L) is: hazel leaf juice 900, and sucrose 300, peptone 100, yeast extract paste 20, sal epsom 10, potassium primary phosphate 10, the pH value is 9;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 50g, yeast extract paste 20g, peptone 20g is dissolved in the 900ml water, is settled to 1000ml, regulates pH to 9;
2) configuration solid-state fermentation culture medium: the 2000g barley is joined in the 1000g nutritive water;
3) solid state fermentation: 75ml secondary liquid seeds is inoculated in step 2) dry weight of gained is on the 150g solid-state fermentation culture medium, and in 40 ℃ of fermentation culture 2 days, bed thickness was 10cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
III, liquid submerged fermentation method are produced the ferfas hypha fluid;
1) configuration liquid nutrient medium: the bright hazel leaf of 100g is put into 1000ml water boil 40 minutes after-filtration, get filtrating, add the sucrose of 50g, the peptone of 10g; The yeast extract paste of 5g, the sal epsom of 1g, the potassium primary phosphate of 1g; Water is settled to 1000ml, and adjust pH is 9, and sterilization promptly gets;
2) fermentation condition is: the secondary liquid seeds 1ml of gained among the I is inoculated in 1) on the liquid nutrient medium 2ml of gained; Liquid amount is a volumetrical 60%, and at 40 ℃, Ventilation Rate is 30 liters/minute; Mixing speed is to cultivate 2 days under 200 rev/mins the condition, the ferfas hypha fluid.
Embodiment 8 (preparation ferfas yellow rice wine)
With the purple rice of 100g and 10g ferfas mycelium powder as raw material, insert 8.8g koji and 5.5g activated yeast mixture after, 20 ℃ of bottom fermentations 10 days, 15 ℃ of standing for fermentation 50 days,, process the ferfas yellow rice wine then according to the making method of yellow rice wine.
Embodiment 9 (preparation ferfas yellow rice wine)
With the wheat of 100g and 200g ferfas hypha fluid as raw material, insert the koji and 3g activated yeast mixture of 6g after, 40 ℃ of bottom fermentations 2 days, 30 ℃ of standing for fermentation 20 days,, process the ferfas yellow rice wine then according to the making method of yellow rice wine.
Embodiment 10 (preparation ferfas yellow rice wine)
With the rice of 100g and 100g ferfas mycelium powder as raw material, insert the koji and 6g activated yeast mixture of 10g after, 30 ℃ of bottom fermentations 6 days, 20 ℃ of standing for fermentation 40 days,, process the ferfas yellow rice wine then according to the making method of yellow rice wine.
Embodiment 11 (preparation ferfas wine)
Embodiment 1 gained ferfas mycelium fermentation powder is used ultrasonic disruption, and 120 ℃ of boilings 1 hour are put to room temperature; Every 100g ferfas mycelium fermentation powder adds yeast-saccharifying enzyme-acetic acid premixed liquid of 50mL; 20 ℃ of ferment at constant temperature 30 days are used filtered through gauze then, and supernatant is got in the 4 ℃ of fermentations 80 days of filtrating, the millipore filtration degerming, and sealing, low tempertaure storage promptly gets; It is 8% Glacial acetic acid min. 99.5 that described yeast-saccharifying enzyme-acetic acid premixed liquid comprises volume(tric)fraction, yeast 15g/100mL, saccharifying enzyme 20g/100mL.
Embodiment 12 (preparation ferfas wine)
The ferfas mycelium fermentation powder of embodiment 2 gained is used ultrasonic disruption, and 120 ℃ of boilings 3 hours are put to room temperature; Every 100g ferfas mycelium fermentation powder adds yeast-saccharifying enzyme-acetic acid premixed liquid of 150mL; 40 ℃ of ferment at constant temperature 10 days are used filtered through gauze then, and supernatant is got in the 8 ℃ of fermentations 20 days of filtrating, the millipore filtration degerming, and sealing, low tempertaure storage promptly gets; It is 3% Glacial acetic acid min. 99.5 that described yeast-saccharifying enzyme-acetic acid premixed liquid comprises volume(tric)fraction, yeast 5g/100mL, saccharifying enzyme 10g/100mL.
Embodiment 13 (preparation ferfas wine)
The ferfas mycelium fermentation powder of embodiment 1 gained is used ultrasonic disruption, and 120 ℃ of boilings 2 hours are put to room temperature; Every 100g ferfas mycelium fermentation powder adds yeast-saccharifying enzyme-acetic acid premixed liquid of 100mL; 30 ℃ of ferment at constant temperature 20 days are used filtered through gauze then, and supernatant is got in the 6 ℃ of fermentations 50 days of filtrating, the millipore filtration degerming, and sealing, low tempertaure storage promptly gets; It is 5% Glacial acetic acid min. 99.5 that described yeast-saccharifying enzyme-acetic acid premixed liquid comprises volume(tric)fraction, yeast 10g/100mL, saccharifying enzyme 15g/100mL.
Claims (6)
1. ferfas wine, it is characterized in that: its raw material comprises the ferfas mycelium of 5-95 weight part, the cereal of 95-5 weight part.
2. ferfas wine according to claim 1, wherein said ferfas is mycelial preparation method may further comprise the steps:
I, strain cultivation:
1) slant strains is cultivated: with wild immature fresh kames, remove surperficial silt, meat bacteria organization's piece of getting middle part 2-3mm with scalper is inoculated in the slant medium, puts into 20-35 ℃ of constant incubator and cultivates 2-30 days, gets slant strains; Described slant culture based formulas is: with peeling potatoes, section was boiled 15-20 minute mistake and was filtered filtrating; The hazel leaf is put into water boiled to cross in about 10-15 minute and filter filtrating, merge above-mentioned two kinds of filtratings, add sucrose again, agar is used the zero(ppm) water constant volume, and adjust pH is 6-9;
2) level liquid seed culture: the broken bacterium piece that is cut into 2-4mm after the slant strains activation of cultivating in the step 1) transferred shaking of level liquid seed culture medium is housed carries out the level liquid seed culture in the bottle; In culture temperature is 15-40 ℃; Rotating speed is on 50-300 rev/min the rotary or reciprocating type bottle swingging machine shaking culture 2-15 days, must contain the level liquid seed of bacterial classification; The prescription of described level liquid seed culture medium is: hazel leaf juice 300-900g/L, and sucrose 1-300g/L, peptone 1-100g/L, yeast extract paste 1-20g/L, sal epsom 1-10g/L, potassium primary phosphate 1-10g/L, the pH value is 6-9;
3) the secondary liquid seeds is cultivated: with step 2) in the level liquid seed cultivated transfer according to the inoculum size of 5-50% volume ratio and shaking of secondary liquid seed culture medium is housed carries out the secondary liquid seeds in the bottle and cultivate; Culture temperature is 15-40 ℃; Rotating speed is on 50-300 rev/min the rotary or reciprocating type bottle swingging machine shaking culture 2-15 days, must contain the secondary liquid seeds of bacterial classification; The prescription of described secondary liquid seed culture medium is: hazel leaf juice 300-900g/L, and sucrose 1-300g/L, peptone 1-100g/L, yeast extract paste 1-20g/L, sal epsom 1-10g/L, potassium primary phosphate 1-10g/L, the pH value is 6-9;
II, solid state fermentation are produced ferfas mycelium fermentation powder:
1) configuration nutritive water: with sucrose 10-50g, yeast extract paste 0-20g, peptone 0-20g, potassium primary phosphate 0-10g, sal epsom 0-10g is dissolved in the 700-900ml water, is settled to 1000ml, regulates pH to 6-9;
2) configuration solid-state fermentation culture medium: by weight for 0.8-2: 1 joins cereal in the nutritive water;
3) solid state fermentation: according to the 1-50% volume/weight of solid fermentation substratum dry weight; The secondary liquid seeds is inoculated in step 2) on the solid-state fermentation culture medium of gained; In 20-40 ℃ of fermentation culture 2-60 days; Bed thickness is 1-10cm, and the mycelium on culture surface is when transferring tawny in vain, and fermentation culture finishes;
4) drying is pulverized, and sieves, and sterilization promptly gets ferfas mycelium fermentation powder.
3. the mycelial preparation method of ferfas according to claim 2, wherein said preparation process also comprises:
The liquid submerged fermentation method is produced ferfas mycelium powder or ferfas hypha fluid;
1) configuration liquid nutrient medium: bright hazel leaf is put into water boil 20-40 minute after-filtration, get filtrating, add sucrose, peptone, yeast extract paste, sal epsom, potassium primary phosphate, the water constant volume, adjust pH is 6-9, sterilization promptly gets;
2) fermentation condition is: according to the inoculum size of 5-50% volume ratio the secondary liquid seeds of gained among the I is inoculated in 1) on the liquid nutrient medium of gained; Liquid amount is volumetrical 60-80%; At 20-40 ℃; Ventilation Rate be the 30-60 liter/minute, mixing speed is to cultivate 2-30 days under 200-800 rev/min the condition, the ferfas hypha fluid;
3) with step 2) gained ferfas hypha fluid is centrifugal, filters, and drying is pulverized, and sieves, and sterilization promptly gets the ferfas mycelium powder.
4. a kind of ferfas wine according to claim 1, adopt following preparation method:
The preparation method one: with weight ratio is 1: the cereal of 0.1-2 and ferfas mycelium powder or ferfas hypha fluid are as raw material; After the koji of access 2-8% weight ratio and the activated yeast mixture of 1-5% weight ratio; At 20-40 ℃ of bottom fermentation 2-10 days; At 15-30 ℃ of standing for fermentation 20-50 days,, process the ferfas yellow rice wine then according to the making method of yellow rice wine; Or
The preparation method two: the ferfas mycelium fermentation powder of claim 2 gained is used ultrasonic disruption, 120 ℃ boiling 1-3 hour, put to room temperature; Every 100g ferfas mycelium fermentation powder adds yeast-saccharifying enzyme-acetic acid premixed liquid of 50-150mL; 20-40 ℃ ferment at constant temperature 10-30 days, use filtered through gauze then, filtrate 4-8 ℃ the fermentation 20-80 days, get supernatant, the millipore filtration degerming, the sealing, low tempertaure storage promptly gets; It is the 3-8% Glacial acetic acid min. 99.5 that described yeast-saccharifying enzyme-acetic acid premixed liquid comprises volume(tric)fraction, yeast 5-15g/100mL, saccharifying enzyme 10-20g/100mL.
5. a kind of ferfas wine according to claim 1 is characterized in that described Tuber bacterial strain is selected from one or more in Chinese ferfas, India truffle, black truffle, white ferfas or the summer truffle.
6. a kind of ferfas wine according to claim 1, wherein said cereal is rice, millet, corn, purple rice, red rice, black rice, one or more of wheat, barley, highland barley, buckwheat or Chinese sorghum.
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