CN104956925A - Production method for inonotus obliquus continuous sumbmerged fermentation liquid and ferment powder - Google Patents
Production method for inonotus obliquus continuous sumbmerged fermentation liquid and ferment powder Download PDFInfo
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Abstract
The invention relates to the technical field of inonotus obliquus, and specifically relates to a production method for inonotus obliquus continuous sumbmerged fermentation liquid and ferment powders. Provided is the production method for inonotus obliquus continuous sumbmerged fermentation liquid and ferment powders, the method combining fermentation facility technology with Chinese herbal medicine. The method comprises the following steps: a, mother spawn purification; b, inonotus obliquus primary fermentation; c, continuous mixing submerged fermentation; d, manufacturing of fermentation broth; e, manufacturing of ferment powders. The method combines the fermentation facility technology with Chinese herbal medicine, the biological activity, nutritional ingredients, and chemical characteristics which the inonotus obliquus strain specially has are combined with the Chinese herbal medicine, so as to further continuously perform sumbmerged fermentation. The method is mainly applied in human body care and treatment.
Description
Technical field
The present invention relates to Inonotus obliquus technical field, more specifically, relate to the production method of the continuous submerged fermentation liquid of Inonotus obliquus and yeast powder.
Background technology
Inonotus obliquus belongs to Basidiomycotina, Hymenomycetes, non-brown Zoopagales, Polyporaceae, brown pore fungi genus, and be a kind of medicinal fungi be born on birch, its fruit body presents the black block-shape morphology being similar to carbon.When Inonotus obliquus played drug effect after 10 ~ 15 years, it can blot the marrow of birch, makes trees withered or died.Inonotus obliquus is mainly distributed in Russia the north, Northern Europe, China Dark Longjiang, Japan (Hokkaido), grows the area of north latitude 40 ° ~ 50 ° on the Northern Hemisphere.Inonotus obliquus chemical composition has polysaccharide, inonotus obliquus, Inonotus obliquus alcohol, multiple oxidation triterpene compound, the acid of bolt bacterium, multiple lanosterol type triterpene compound, folic acid derivatives, aromatic vanillic acid, syringic acid and γ hydroxybenzoic acid, reports in addition and has isolated tannin compound, steroids, alkaloid compound, melanin class, low molecule Polyphenols and lignin compound. and Inonotus obliquus contains a large amount of anticancer, hypotensive, hypoglycemic, vegetable fiber class polysaccharide body of bringing back to life immunization.Can improve the vigor of immunocyte, inhibition cancer cell diffusion and recurrence, prevent the absorption of the harmful substances such as carcinogen in stomach and intestine, and promote excretion.The refining bacterium powder of Inonotus obliquus reaches 93% to diabetes cure rate.
Summary of the invention
The invention provides the production method of a kind of continuous submerged fermentation liquid of Inonotus obliquus of fermentation installation technology and medium-height grass being combined and yeast powder.
The technical solution used in the present invention is:
The production method of the continuous submerged fermentation liquid of Inonotus obliquus and yeast powder, carry out according to following steps:
A, female kind purifying;
B, Inonotus obliquus primary fermentation;
C, mix submerged fermentation continuously;
The preparation of d, zymotic fluid;
The preparation of e, yeast powder.
Described a step is carried out in such a way: 1) preparation raw material; 2) medium is made; 3) packing test tube; 4) Sai Pisai; 5) test tube wrapping; 6) medium sterilization; 7) inclined-plane is put; 8) sterilization effect is checked; 9) female kind is inoculated; 10) inonotus obliquus mother seed kind is cultivated;
Above-mentioned preparation raw material is specially: be configured respectively according to the following component of mass percent, Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, the water of magnesium sulfate 0.05% and surplus, PH is 7;
Above-mentioned making medium is specially: Inonotus obliquus sporophore wears into 80 object fine powders, Inonotus obliquus sporophore and potato are added in the water of 1000ml, boil rear low baking temperature and keep 30 minutes, then by double-deck adhesional wetting filtered through gauze, add agar afterwards and continue heating, and continuous glass bar is stirred to agar dissolves completely, add glucose, potassium dihydrogen phosphate, peptone, magnesium sulfate successively again, continue to heat and be stirred to all to dissolve, finally complement to 1000 milliliters with clear water, complete medium and make.
Described b step is carried out in such a way: 1) preparation raw material; 2) culture fluid makes and sterilizing; 3) culture fluid inoculation; 4) cultivation of liquid spawn;
Above-mentioned preparation raw material is specially: be configured respectively according to the following component of mass percent, Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, corn flour 5%, wheat bran 3%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, the water of magnesium sulfate 0.05% and surplus, PH is 7;
Above-mentioned culture fluid makes and is specially with sterilizing: Inonotus obliquus sporophore wears into 80 object fine powders, Inonotus obliquus sporophore and potato are added in the water of 1000ml, boil rear low baking temperature and keep 30 minutes, then by double-deck adhesional wetting filtered through gauze, add agar afterwards and continue heating, and continuous glass bar is stirred to agar dissolves completely, add glucose, potassium dihydrogen phosphate, corn flour, wheat bran, peptone, magnesium sulfate successively again, continue to heat and be stirred to all to dissolve, finally complement to 1000 milliliters with clear water, complete culture fluid and make; To be divided in fermentor after culture fluid boils, charge weight is 60% of population size, then with sealing paper or sealed membrane, utensil mouth package is lived, packaged liquid appliance ad is loaded in pressure cooker and carries out autoclaving, treat that temperature rises to 121 DEG C and starts timing, temperature controls at 121-123 DEG C, maintains and can reach sterilization effect in 30 minutes.
Described step c is carried out in such a way: 1) continuously ferment liquid nutrient medium; 2) sterilization of continuous fermentation tank; 3) charging sterilizing; 4) inoculate; 5) cultivation of fermented liquid;
The above-mentioned liquid nutrient medium that continuously ferments is specially: form according to the following component of mass percent, Inonotus obliquus sporophore dry powder 2%, Portugal's glucose 0.5%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.075%, potato concentrate 5%, peptone 0.05%, defoamer 60mL/100L, 50 object corn flour 3%, wheat bran 5%, Herba Ecliptae 0.15%, the raw bighead atractylodes rhizome 0.15%, radix bupleuri 0.1%, stir-baked FRUCTUS GARDENIAE 0.1%, moutan bark 0.15%, the fruit of glossy privet 0.15% processed, Poria cocos 0.15%, cogongrass rhizome 0.15%, grow radix paeoniae alba 0.15%, fructus viticis 0.1%, prepared rhizome of rehmannia 0.15%, Radix Codonopsis 0.15%, reed rhizome 0.15%, the water composition of stir-baked SEMEN ARMENIACAE AMARUM 0.1% and surplus.
Described Step d is carried out in such a way: use filter to start to filter temperature 25 ~ 28 DEG C, reflux and filter liquid 10 minutes, collects the filtrate of clarification, obtain Inonotus obliquus zymotic fluid; Filter the liquid of gained through super high temperature instantaneous sterilizing machine sterilization, sterilization temperature (110 ± 5) DEG C, 10 seconds time, sterilized liquid is put and is cooled to 50 ~ 55 DEG C rapidly to temporary storage tank, concentrates; Concentrated under vacuum 0.096 ~ 0.099 MPa, fluid temperature 50 ~ 55 DEG C, steam pressure 0.196 ~ 0.294 MPa, be concentrated into after soluble solid reaches 42% ~ 45% and carry out re-pasteurization; Liquid is loaded in special-purpose bottle (bag) by the liquid liquid-filling machine concentrated, then 90 DEG C of sterilizations 30 minutes; By finished product constant refrigeration in-18 ~-25 DEG C of freezers.
Described step e is carried out in such a way: after the whole drying of fermentation solid content low-temperature freeze-dry equipment after filtration, then wear into 120 object fine powders with low temperature milling equipment, obtain Inonotus obliquus yeast powder, then carry out vacuum packaging preservation.
The beneficial effect that the present invention has is: fermentation installation technology and medium-height grass combine by the present invention, with the distinctive biologically active of its Inonotus obliquus bacterial classification itself and nutrient component and chemical property in conjunction with Chinese herbal medicine submerged fermentation continuously further, the liquid produced and solid content have balance blood sugar, reduce high fat of blood, hypertension, improve angiocardiopathy, improve gastrointestinal function adjusting blood lipid, analgesia, removing toxic substances, antiallergy, improve blood oxygen ability, strengthen the activity of brain cell, strengthen memory, anti-tampon, vascular sclerosis and apoplexy and improve dementia symptom.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
Technological process of the present invention is carried out in the following manner: 1) female kind purifying; 2) Inonotus obliquus primary fermentation; 3) submerged fermentation is mixed continuously; 4) preparation of zymotic fluid; 5) preparation of yeast powder.
One, female kind purifying
1, fill a prescription: 1000ml is example
Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, magnesium sulfate 0.05%, PH7.
2, medium makes
Select the potato of high-quality, peeling, cuts out eye, reams dark green part, and be cut into the thin slice of long 0.5 millimeter, take 200 grams, Inonotus obliquus sporophore will wear into 80 object fine powders in advance.Add water 1 000 milliliters, boil rear low baking temperature and keep 30 minutes, potato is boiled and not crisp, i.e. 8 ripe left and right.Then by 2 layers of adhesional wetting filtered through gauze, add agar afterwards and continue heating, and continuous glass bar is stirred to agar dissolves completely, finally adds glucose, potassium dihydrogen phosphate, peptone, magnesium sulfate etc. successively, continuation is heated and is stirred to and all dissolves.Finally complement to 1000 milliliters with clear water, can packing be carried out.
3, packing test tube.Left hand holds 5 test tube mouths of pipe upward, and lower end is neat, above moves test tube to funnel flexible pipe, inserts in vitro about 3 centimetres.The right hand clutches flexible pipe flow-stopping clip, makes medium liquid flow into test tube, and often prop up that test tube loading amount is about test tube length 1/5.Now unclamp flow-stopping clip liquid to stop flowing into.Flexible pipe is moved away by under test tube, then packing the 2nd test tube, other test tubes of Using such method packing, until all install.Packing test tube requires that speed wants fast, especially at low temperature season, to prevent culture medium solidifying.Loading amount is wanted accurately, because the loading amount inclined-plane be put on the high side shortens, waste medium again, charge weight is on the low side, and make the inclined-plane that is put into too thin, nutrition is few, and mycelia is difficult to normal growth.Note avoiding medium to be attached to the test tube mouth of pipe, in order to avoid cause the phenomenon of adhesion skin plug, also easily cause skin to multiply miscellaneous bacteria beyond the Great Wall, leave hidden danger to strain quality.
4, Sai Pisai, fills in test tube with hand special skin plug, and skin plug will be close to the wrinkle resistant line of tube wall, and the skin plug part filled in pipe is about 2.5 centimetres, and exposed parts accounts for 1/3 of Pi Sai overall length.
5, test tube wrapping.Test tube is stoppered Pi Saihou, gets the test tube of 10 same specifications, is encased by tampon, and extends downwardly in the middle part of test tube, be bundled into one with bonnet cotton with brown paper or double-deck newspaper.
6, medium sterilization.Mother culture Quito adopts portable high-pressure sterilizing pot to carry out sterilizing, and sterilization process is: flexibly whether the manometer of inspection high-pressure steam sterilizing pan and each position, reliably; Especially note checking whether safety valve blocks or become rusty dead.In pot, add about 3 kilograms of running water, add to support lower edge.The medium tampon tied upwards vertically is put into bacterium basket (bucket), and inserted in the wireway of sleeve lining by the blast pipe that pot cover hangs down, then both hands are exerted oneself simultaneously, respectively by two of diagonal screw tightenings.Close air bleeding valve after capping and start heating, when being heated to pressure and reaching 0.05 MPa, open air bleeding valve, drain cold air, after Pressure Drop to 0, can air bleeding valve be closed.Preferably arrange 2 times according to upper method.Continue heating, start timing when pressure increase to 0.11 MPa, keep 30 minutes under 0.11 ~ 0.14 MPa, stop heating, make it cool voluntarily and be down to 0 DEG C.Open air bleeding valve, then pot door is opened the gap heat radiation of 10 cm.
7, inclined-plane is put.After observing newspaper drying, open pot cover, take out in time when medium temperature drops to about 60 DEG C and put inclined-plane.Notice that medium will cover bottom test tube, chamfer length should with 3/5ths of test tube length.When in vitro medium is packed up for subsequent use after solidifying completely.The medium prepared should be placed on aeration-drying place and preserve.If just put inclined-plane when temperature is very high, culture medium solidifying rear surface there will be more condensed water, unfavorable mycelial growth.After being well placed inclined-plane, cover medium with abundant cotton, medium slow cooling is cooled, also can reduce condensed water and occur.
8, sterilization effect inspection.Get part test tube slant at random, carry out blank at being placed on 28 DEG C and cultivate, after 3 days, check on inclined-plane with or without bacterium and mold colony.If find that there is miscellaneous bacteria, illustrate that sterilizing is not thorough, sterilizing again.
9. female kind is inoculated.Before inoculation, medium is put into inoculating hood to carry out disinfection, sterilization requires to be undertaken by aforementioned.During inoculation, hand and inoculation hook with 75% cotton ball soaked in alcohol wipe, light alcolhol burner, flame periphery space is made to form aseptic area, left hand is parallel picks up female kind test tube and the slant tube for inoculation side by side, and upwards, the mouth of pipe will flush in two test tube slants, right hand thumb and forefinger hold inoculation hook, sterilizing that the flame of alcolhol burner burns.The part of test tube that inoculation hook is allowed for access all will be carried out calcination.Left hand test tube is moved to by flame, right-handed little finger of toe, the third finger and palm, under the other folder respectively of flame, the tampon of two test tubes, bakes on flame a little by test tube mouth, to kill the miscellaneous bacteria on the mouth of pipe, subsequently the mouth of pipe is moved to apart from flame 1 ~ 2 centimeters, the inoculation of calcination rake is stretched in strain tube, first contacts inboard wall of test tube, make it cool, the aerial hyphae hook of being climbed wall again falls, and is cut off by medium front end thinnest part, goes out outside pipe together with aerial hyphae in the lump hook.With inoculation hook (shovel, rake), mother's kind is slit into many blockages, then picking one fritter in length and breadth again, moves into rapidly the middle front part of the test tube slant of inoculation, extract inoculation hook gently out, roasting test tube mouth once, crosses rapidly the tampon of flame beyond the Great Wall again, and a mother plants generally can expand and connects 30 ~ 40, test tube.
Technical essential: whole process must carry out sterile working on flame side, the inoculation block in female kind and aging will the removing of periphery, the mycelia block selecting cell age short is produced.Before test tube takes out from inoculating hood, should be labelled by the top of a tube facing side, write bacterium numbering, inoculation date exactly.
10. the cultivation of inonotus obliquus mother seed kind
Inoculate rear test tube is put in incubator (room) and carry out heat insulating culture.Temperature is adjusted to 25 DEG C, and within general 15 ~ 20 days, mycelium can cover with test tube, and because Inonotus obliquus mycelial growth rate is comparatively slow, so 2 bacterium blocks of can transferring on 1 female kind inclined-plane, the time of covering with test tube can shorten half.
The mother of 11. Inonotus obliquus plants quality standard
Require that test tube is complete, without damaged, tampon is dry, clean, degree of tightness is suitable for, and can meet ventilative and bacteriological filtration requirement; Beveled top end is apart from tampon 45 centimetres; White mycelium or micro-yellow, robust growth, dense, even, colony edge is neat; Medium is drying shrinkage not, without blackening, without miscellaneous bacterias such as green mold bacterium.
The reason of 12. inonotus obliquus mother seed kind growth failures and solution
(1) around inoculation block, mycelial growth is unable, even becomes without mycelia region.This is because inoculation block is bigger than normal, provenance cell age is not suitable for, cultivation temperature is too high, medium uses caused by the reasons such as formula, a Medium's PH Value >8 continuously.Processing method is: inoculation block is got smaller as far as possible, and select of the right age provenance, regulation culture temperature is about 24 DEG C, replaced medium formula, and several formula is used alternatingly, and makes the pH value after medium sterilization reach about 7.2.
(2) in vitro aerial hyphae there is the kermesinus globule.This be due to cultivation temperature too high caused by.Treating method is regulation culture temperature to 22 ~ 24 DEG C.
(3) mycelia lodging, jaundice, aerial hyphae obviously grows irregular, and branch is unintelligible, grows unable, dead mycelia.This is because provenance is aging, inoculation time medium is chosen too thick, cultivation temperature is too high or caused by the reason such as excessive temperature differentials, culture environment improper ventilation, fungi serious uneven for hypoxgia, the nutrition of medium ratio or virus infections.
Two, Inonotus obliquus primary fermentation
1, fill a prescription: 50000mL is example
Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, corn flour 5%, wheat bran 3%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, magnesium sulfate 0.05%, PH7.
2. culture fluid makes and sterilizing
Boil culture fluid according to making female method of planting solid culture medium, will be divided in after culture fluid boils in fermentation fermenting device, charge weight is 60% of population size, is then lived by utensil mouth package with special sealing paper or sealed membrane.Loaded in pressure cooker by packaged liquid appliance ad and carry out autoclaving, treat that temperature rises to 121 DEG C and starts timing, temperature controls at 121-123 DEG C, maintains and can reach sterilization effect in 30 minutes.
3, culture fluid inoculation
The fermented liquid utensil after sterilizing at special inoculation indoor inoculation, its method chooses new cultured bacterial classification one (need shift to an earlier date activation in 24 hours if take out from refrigerator), loosen the sealing packing material of utensil, convenient in order to inoculation, tampon is turned clockwise loosening on alcolhol burner flame side, be placed on one side, then on alcolhol burner side, dull and stereotyped sealed membrane is torn, pick up inoculation be hooked in flame burns red, and calcination of slowly tilting, during inoculation, plate lid is opened and be placed on one side, (middle inoculation block does not include inoculation part in) the average inoculation block that dull and stereotyped bacterial classification carries out rule is 0.5*0.5cm, after line, inoculation hook is placed on rack for test tube temporarily.The right hand is extracted in triangular flask tampon is held in the hand, and with inoculation hook picking 4-5 block inoculation block, puts into utensil rapidly.
4, the cultivation of liquid spawn:
A, postvaccinal liquid appliance ad are placed on shaking table or on magnetic stirring apparatus and cultivate, and temperature controls at 23-24 DEG C, rotating speed or frequency 126/ minute.
Culture fluid color, concentration and pellet form are observed in B, timing.
C, liquid cell age general control were at 7 days, and now occur a large amount of particulate bacterium ball in culture fluid, culture fluid is brown, with fragrant and sweet taste.
Three, submerged fermentation is mixed continuously
1, continuously ferment liquid nutrient medium
Inonotus obliquus sporophore dry powder 2%, Portugal's glucose 0.5%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.075%, potato concentrate 5%, peptone 0.05%, defoamer 60mL/100L, 50 object corn flour 3%, wheat bran 5%, Herba Ecliptae 0.15%, the raw bighead atractylodes rhizome 0.15%, radix bupleuri 0.1%, stir-baked FRUCTUS GARDENIAE 0.1%, moutan bark 0.15%, the fruit of glossy privet processed 0.15%, Poria cocos 0.15%, cogongrass rhizome 0.15%, grow radix paeoniae alba 0.15%, fructus viticis 0.1%, prepared rhizome of rehmannia 0.15%, Radix Codonopsis 0.15%, reed rhizome 0.15%, stir-baked SEMEN ARMENIACAE AMARUM 0.1%.
2, the sterilization of continuous fermentation tank
Sterilizing feed supplement tank is added water to 70% of cumulative volume.Open feed supplement tank power switch, temperature is set to 121 DEG C-125 DEG C, after getting to this temperature, close air header road valve, open steam valve, allow steam enter three grades, level Four successively by secondary filter, finally arrive fermentation tank, after fermentation jar temperature reaches ll8 DEG C-120 DEG C, valve, sample valve will be inoculated, air bleeding valve is opened a bit, steam is allowed slowly to discharge a bit, sterilizing (will arrange the condensed water of a filter bottom generation for every 3-5 minutes in whole process) is carried out to it, keep after 30 minutes, first open fermentation tank drain tap below, then sterilizing feed supplement tank discharging opening valve is slowly opened a bit, high-temperature high pressure water is allowed to flow out a little, after its threeway preheating, open the steam valve entering punishment in advance pipeline, after drain tap after fermentation tank has a large amount of steam to discharge, close fermentation tank drain tap below, open fermentation tank charging aperture valve below, then sample valve is closed, the rapid metal tube head that will sample inserts in formaldehyde bottle, close inoculation and drain tap on fermentation tank deck, first open inoculation and rob controlled valve, again fermentation tank is gone out kind of a mouth valve slowly to open a bit, high-temperature high pressure water is allowed to flow out a little, after its threeway preheating, open the steam valve entering seed output pipe road, when inoculation snatch away kind of a mouth have a large amount of steam discharge after, gas output is turned down, have a small amount of steam to discharge, sterilizing is closed inoculating gun switch and is gone out bacterial classification mouth valve after 30 minutes, for subsequent use with aseptic pouch seal.Temperature be set to lower than room temperature, open air pump, close steam sub valve, open air total valve, dry up filter core, allow filtrated air enter in tank, adjustment tank pressure, at 0.03-0.05MPa, closes the drain tap below filter, allows whole system keep malleation.With 75% medicinal alcohol cotton balls, the inoculation mouth valve of fermentation tank top center is sealed after terminating.
3, charging sterilizing
First sterilizing feed supplement tank steam total valve and discharging opening valve is closed, the water (if the too poor need of water quality carry out purified treatment) of required loading amount 70% will be added in feed supplement tank, open sterilizing feed supplement tank power switch, temperature is set to 95 DEG C, all raw materials with hot water dissolving together, when in feed supplement tank, temperature is raised to 950 DEG C, open discharging opening valve and pass into a small amount of air, first defoamer is added under the stirring of air, when feed temperature is close to 95 DEG C, close discharging opening valve, after more than 95 DEG C, close all valves on feed supplement tank; Temperature is set to 121 DEG C, opens timer (sterilization time sets 30 minutes); Connected by heat exchanger circulating water pipeline, after reaching sterilising temp, by the charge door valve at sterilization tank top, drain tap is all opened a bit, allows steam discharge a little, to its sterilizing, after reaching sterilization time, closes heating power supply and all valves in sterilization tank top.First discharge 500mL-1000mL feed liquid from the sewage draining exit below sterilizing feed supplement tank, again down cycles water is led to, then open feed supplement tank discharging opening valve, by the composts or fertilisers of cultivating of bacterium of having gone out, enter fermentation tank after being lowered the temperature by heat exchanger, sterilizing temperature controller temperature is controlled below 25 DEG C.
4, inoculate
Within 2 hours, with ozone, all for culturing room environment are carried out disinfection in advance, then cultured shake-flask seed is taken out, from inoculation valve, bacterial classification is accessed under Alcohol Flame protection,--as two people coordinate carry out, be enclosed within inoculation mouth with the fire ring soaking alcohol, close air intake valve, open drain tap, allow pressure zero in fermentation tank, the firstly light fire ring, the cotton ball soaked in alcohol of inoculation valve port is taken away with tweezers, open inoculation valve, the hand-held shaking flask of second people, metal pipe mouth is put into flame, the first tweezers pull out the tampon in tube head, pipe inserts in inoculation valve by the second people, tiltedly carry shaking flask access bacterial classification, pipe is extracted after having connect, valve-off, remove fire ring, inoculate salty new use 75% medical cotton ball to seal, open air intake valve, pass into filtrated air, fermented and cultured.
5, the cultivation of zymotic fluid
With the impeller speed stir culture 72 hours of 180 revs/min after inoculation, middlely detect sugar, nitrogen, pH value, liquid asepsis degree every sampling in 12 hours.Putting tank standard is that zymotic fluid pH value drops to 5, and residual sugar content is down to about 2.5, and ammoniacal nitrogen is no more than 30 mg/litre, and bacterium ball concentration reaches 1000 ~ 1500/milliliter, or through 3000 revs/min centrifugal 10 points, solid content weight in wet base is at 20 ~ 25 grams/100 milliliters.Check that zymotic fluid qualitative items comprises: purity test, microscopy or slat chain conveyor are without bacterium, mould; Vigor checks, microscopy bacterium ball edge mycelia branch is fine and closely woven, and color depth with during violet staining, hyphal cell protoplasm not yet occurs agglutination phenomenon, static 5 points of mash, and solid content does not sink; Bacterium ball size, 80% bacterium bulb diameter is less than 1 millimeter; Temperature controller temperature is set to 23-25 DEG C, and in tank, pressure keeps 0.03-0.04MPa, throughput: material: gas is in 1:(0.2-0.5) v/v ferments 10 days.
Four, Inonotus obliquus fluid separation applications filters (preparation of zymotic fluid)
1, use special filtering machine to start to filter temperature 23 ~ 25 DEG C, require that filter screen reaches 120 orders, reflux and filter liquid 10 minutes, collect the filtrate of clarification.Obtain Inonotus obliquus zymotic fluid.
2, a sterilization: filter the liquid of gained through super high temperature instantaneous sterilizing machine sterilization, sterilization temperature (110 ± 5) DEG C, 10 seconds time.Sterilized liquid is put and is cooled to 50 ~ 55 DEG C rapidly to temporary storage tank, concentrates.
3, Vacuum Concentration: concentrated under vacuum 0.096 ~ 0.099 MPa, fluid temperature 50 ~ 55 DEG C, steam pressure 0.196 ~ 0.294 MPa, is concentrated into after soluble solid reaches 42% ~ 45% and carries out re-pasteurization.
4, re-pasteurization: liquid is loaded in special-purpose bottle (bag) by the liquid liquid-filling machine concentrated, then 90 DEG C of sterilizations 30 minutes.
5, refrigerate: by finished product constant refrigeration in-18 ~-25 DEG C of freezers.
6, product quality indicator
(1). organoleptic indicator
Color and luster: pitchy liquid is translucent; Flavour and smell: there is the distinctive fragrance of Inonotus obliquus and smell, without being charred taste when being diluted to Normal juice concentration, free from extraneous odour; Tissue morphology: be organized as translucent uniform liquid, five layerings, deposited phenomenon; Impurity: without exogenous impurity.
(2). physical and chemical index
Soluble solid 42% ± 1%; PH value 5.0 ~ 7.0.
(3). microbiological indicator
Total number of bacteria≤3000/milliliter; Coliform, pathogenic bacteria must not detect.
Five, solid content freeze-drying abrasive dust (preparation of yeast powder) is filtered
1, be laid in lyophilized plate by Inonotus obliquus filtering fermentation liquor solid, should keep its high uniformity when thing is laid in lyophilized plate in filter, to reduce batch interpolation of water content after freeze-drying, height can not be greater than 2cm, in order to avoid affect effect;
2, treat in lyophilized plate that lyophilized products height can not lower than 1.5cm;
3, open freeze dryer, ruuning situation sees the following form:
4, after the whole drying of fermentation solid content low-temperature freeze-dry equipment after filtration, then wear into 120 object fine powders with low temperature milling equipment, then carry out vacuum packaging preservation.
Claims (6)
1. the production method of the continuous submerged fermentation liquid of Inonotus obliquus and yeast powder, is characterized in that, carry out according to following steps:
A, female kind purifying;
B, Inonotus obliquus primary fermentation;
C, mix submerged fermentation continuously;
The preparation of d, zymotic fluid;
The preparation of e, yeast powder.
2. the production method of the continuous submerged fermentation liquid of Inonotus obliquus according to claim 1 and yeast powder, is characterized in that, described a step is carried out in such a way: 1) preparation raw material; 2) medium is made; 3) packing test tube; 4) Sai Pisai; 5) test tube wrapping; 6) medium sterilization; 7) inclined-plane is put; 8) sterilization effect is checked; 9) female kind is inoculated; 10) inonotus obliquus mother seed kind is cultivated;
Above-mentioned preparation raw material is specially: be configured respectively according to the following component of mass percent, Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, the water of magnesium sulfate 0.05% and surplus, PH is 7;
Above-mentioned making medium is specially: Inonotus obliquus sporophore wears into 80 object fine powders, Inonotus obliquus sporophore and potato are added in the water of 1000ml, boil rear low baking temperature and keep 30 minutes, then by double-deck adhesional wetting filtered through gauze, add agar afterwards and continue heating, and continuous glass bar is stirred to agar dissolves completely, add glucose, potassium dihydrogen phosphate, peptone, magnesium sulfate successively again, continue to heat and be stirred to all to dissolve, finally complement to 1000 milliliters with clear water, complete medium and make.
3. the production method of the continuous submerged fermentation liquid of Inonotus obliquus according to claim 1 and yeast powder, it is characterized in that, described b step is carried out in such a way: 1) preparation raw material; 2) culture fluid makes and sterilizing; 3) culture fluid inoculation; 4) cultivation of liquid spawn;
Above-mentioned preparation raw material is specially: be configured respectively according to the following component of mass percent, Inonotus obliquus sporophore 0.5%, potato 5%, glucose 0.05%, corn flour 5%, wheat bran 3%, peptone 0.1%, agar 0.4%, potassium dihydrogen phosphate 0.012%, the water of magnesium sulfate 0.05% and surplus, PH is 7;
Above-mentioned culture fluid makes and is specially with sterilizing: Inonotus obliquus sporophore wears into 80 object fine powders, Inonotus obliquus sporophore and potato are added in the water of 1000ml, boil rear low baking temperature and keep 30 minutes, then by double-deck adhesional wetting filtered through gauze, add agar afterwards and continue heating, and continuous glass bar is stirred to agar dissolves completely, add glucose, potassium dihydrogen phosphate, corn flour, wheat bran, peptone, magnesium sulfate successively again, continue to heat and be stirred to all to dissolve, finally complement to 1000 milliliters with clear water, complete culture fluid and make; To be divided in fermentor after culture fluid boils, charge weight is 60% of population size, then with sealing paper or sealed membrane, utensil mouth package is lived, packaged liquid appliance ad is loaded in pressure cooker and carries out autoclaving, treat that temperature rises to 121 DEG C and starts timing, temperature controls at 121-123 DEG C, maintains and can reach sterilization effect in 30 minutes.
4. the production method of the continuous submerged fermentation liquid of Inonotus obliquus according to claim 1 and yeast powder, it is characterized in that, described step c is carried out in such a way: 1) continuously ferment liquid nutrient medium; 2) sterilization of continuous fermentation tank; 3) charging sterilizing; 4) inoculate; 5) cultivation of fermented liquid;
The above-mentioned liquid nutrient medium that continuously ferments is specially: form according to the following component of mass percent, Inonotus obliquus sporophore dry powder 2%, Portugal's glucose 0.5%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.075%, potato concentrate 5%, peptone 0.05%, defoamer 60mL/100L, 50 object corn flour 3%, wheat bran 5%, Herba Ecliptae 0.15%, the raw bighead atractylodes rhizome 0.15%, radix bupleuri 0.1%, stir-baked FRUCTUS GARDENIAE 0.1%, moutan bark 0.15%, the fruit of glossy privet 0.15% processed, Poria cocos 0.15%, cogongrass rhizome 0.15%, grow radix paeoniae alba 0.15%, fructus viticis 0.1%, prepared rhizome of rehmannia 0.15%, Radix Codonopsis 0.15%, reed rhizome 0.15%, the water composition of stir-baked SEMEN ARMENIACAE AMARUM 0.1% and surplus.
5. the production method of the continuous submerged fermentation liquid of Inonotus obliquus according to claim 1 and yeast powder, it is characterized in that, described Step d is carried out in such a way: use filter to start to filter temperature 25 ~ 28 DEG C, reflux and filter liquid 10 minutes, collect the filtrate of clarification, obtain Inonotus obliquus zymotic fluid; Filter the liquid of gained through super high temperature instantaneous sterilizing machine sterilization, sterilization temperature (110 ± 5) DEG C, 10 seconds time, sterilized liquid is put and is cooled to 50 ~ 55 DEG C rapidly to temporary storage tank, concentrates; Concentrated under vacuum 0.096 ~ 0.099 MPa, fluid temperature 50 ~ 55 DEG C, steam pressure 0.196 ~ 0.294 MPa, be concentrated into after soluble solid reaches 42% ~ 45% and carry out re-pasteurization; Liquid is loaded in special-purpose bottle (bag) by the liquid liquid-filling machine concentrated, then 90 DEG C of sterilizations 30 minutes; By finished product constant refrigeration in-18 ~-25 DEG C of freezers.
6. the production method of the continuous submerged fermentation liquid of Inonotus obliquus according to claim 1 and yeast powder, it is characterized in that, described step e is carried out in such a way: after the whole drying of fermentation solid content low-temperature freeze-dry equipment after filtration, 120 object fine powders are worn into again with low temperature milling equipment, obtain Inonotus obliquus yeast powder, then carry out vacuum packaging preservation.
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