CN105733971A - Saccharomyces cerevisiae strain as well applications and culture method thereof - Google Patents
Saccharomyces cerevisiae strain as well applications and culture method thereof Download PDFInfo
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Abstract
The invention aims at providing a novel saccharomyces cerevisiae strain as well applications and a culture method of the saccharomyces cerevisiae strain. According to the saccharomyces cerevisiae strain as well the applications and the culture method, rhizoma dioscoreae starch in rhizoma dioscoreae zingiberensis can be converted into alcohol, so that the purpose of preparing diosgenin through extraction in a self-sufficiency manner is achieved, and further, the problems that in the prior art, the common commercial yeast can not convert rhizoma dioscoreae starch in rhizoma dioscoreae zingiberensis into alcohol, consequently, the diosgenin producing cost is high, the comprehensive utilization ratio is low, the resources are wasted, and the pollution is serious, are solved.
Description
Technical field
The present invention relates to brewing fermentation technical field, especially relate to a kind of novel Wine brewing yeast strain and application thereof and cultural method.
Background technology
Diosgenin (diosgenin) is the aglucon of dioscin in yam, it has the multiple pharmacological effect such as antiinflammatory, antibacterial, anticancer and blood fat reducing, the primary raw material of synthesizing steroid hormone medicine especially, accounts for the 70% of required steroid total amount.Diosgenin is generally prepared from by Dioscorea zingiberensis, consumption is made in order to what reduce strong acid in preparation process, reduce acid waste water pollutant emission etc., current environment-friendlyenergy is extracted for the method many employings ethanol (ethanol) of diosgenin, it is necessary to ethanol (ethanol) amount of consumption is relatively big, relatively costly, and Dioscorea zingiberensis generally can only extract the diosgenin of 10%, all the other compositions are all used as waste material and are discarded, and cause great waste, making a low multiple use of Dioscorea zingiberensis.
Under normal circumstances, the dioscin in Dioscorea zingiberensis " is wrapped up " layer by layer by substantial amounts of starch and cellulose and, and when polysaccharide such as having substantial amounts of starch, cellulose exists, can disturb the acidic hydrolysis of Rhizoma Dioscoreae protosaponin class, and reaction is not easily complete, and productivity is low.In order to release the starch " parcel " to Saponin, existing generally starch is all adopted to be directly separating, Starch Hydrolysis become glucose after separate produce glucose or other, starch is consumed by aging process, all there is wretched insufficiency in these methods, some starch separation are not thorough, and what have cannot be used for commercial production, and what have wastes starch resource.It is not efficiently used additionally, due to starch and results in substantial amounts of polluter, reduce the comprehensive utilization ratio of Rhizoma Dioscoreae.Owing to commercial yeast of the prior art all cannot be survived in Dioscorea zingiberensis sugar liquid, therefore in the urgent need to a kind of can be the Wine brewing yeast strain of ethanol by Rhizoma Dioscoreae Starch Conversion, with realize by Rhizoma Dioscoreae Starch Conversion for ethanol be conducive to extract diosgenin.
Summary of the invention
It is an object of the invention to provide a kind of Wine brewing yeast strain and application thereof, can being ethanol by the Rhizoma Dioscoreae Starch Conversion in Dioscorea zingiberensis and then be conducive to extracting and produce diosgenin, solving general commercial yeast in prior art cannot be the problem that ethanol causes producing that diosgenin cost is high, makes a low multiple use, the wasting of resources, environmental pollution are serious by the Rhizoma Dioscoreae Starch Conversion in Dioscorea zingiberensis.
This invention address that the technical scheme is that present invention of technical problem provides a kind of novel Wine brewing yeast strain (Saccharomycescerevisaie), its deposit number is CGMCCNo.9518.
The Wine brewing yeast strain of the present invention prepares the application in ethanol in production, it is possible to for preparing ethanol by Dioscorea zingiberensis production, may be used for preparing ethanol by Rhizoma Dioscoreae Starch Production or may be used for preparing ethanol by Dioscorea zingiberensis sugar liquid fermenting and producing.
The cultural method of the Wine brewing yeast strain of the present invention, comprises the steps:
S1, bacterial strain cultivation preserve: being heated by strain cultures, agar powder injects in culture dish and test tube after dissolving, cooling after 120 DEG C of-130 DEG C of autoclavings 30-120 minute, makes flat board and slant tube；Aseptically, in inoculation to plating medium, cultivating 3-4 days at 28-32 DEG C, the bigger colony inoculation of picking, to slant tube, was cultivated then through 3-4 days, and slant tube puts into cold compartment of refrigerator cryopreservation；
S2, bacterial strain activation culture obtain yeast wine: be aseptically inoculated in the liquid tube equipped with 10ml yeast wine culture fluid by the bacterial strain preserved in step S1, when 30-32 DEG C, after cultivating 10-16 hour, then it is transferred in the liquid tube equipped with 10ml yeast wine culture fluid and continues cultivation 10-16 hour.
In the cultural method of the present invention, in step S1, the formula of strain cultures is: Dioscorea zingiberensis sugar liquid 450ml-550ml, MgSO40.01g-0.02g, Co (NH2)20.17g-0.18g, (NH4)2HPO40.12g-0.13g, agar powder is 9-11g.
In the cultural method of the present invention, the formula of the yeast wine culture fluid in step S2 is: Dioscorea zingiberensis sugar liquid 450ml-550ml, MgSO40.01g-0.02g, Co (NH2)20.17g-0.18g, (NH4)2HPO40.12g-0.13g。
In the cultural method of the present invention, also include step S3 upon step s 2, the yeast wine obtained in step S2 for seed by inoculum concentration 8%-15% constantly amplification culture step by step, until required yeast wine amount.
In the cultural method of the present invention, in step s3 in the process of yeast wine amplification culture step by step, including the step A that the little yeast wine for producing is cultivated, its medium and small yeast wine culture fluid formula is Dioscorea zingiberensis sugar liquid 150L-240L, MgSO45g-7g, Co (NH2)260g-70g, (NH4)2HPO440g-60g；Condition of culture: cultivation temperature 28 DEG C-30 DEG C, incubation time 8-12 hour, per hour logical filtrated air 1-5 minute.
In the cultural method of the present invention, in step s3 in the process of yeast wine amplification culture step by step, including the step A that the big yeast wine for producing is cultivated, wherein big yeast wine culture fluid formula is Dioscorea zingiberensis sugar liquid 1500L-2400L, MgSO415g-25g, Co (NH2)2120g-200g, (NH4)2HPO4100g-180g；Condition of culture: cultivation temperature 30 DEG C-32 DEG C, incubation time 8-12 hour, per hour logical filtrated air 1-5 minute.
In the cultural method of the present invention, described Dioscorea zingiberensis sugar liquid is prepared as follows: take Dioscorea zingiberensis by pulverizing and/or grind to form the screened stock of particle diameter≤1mm, amylase is added by the amount of 8u/g-16u/g starch, liquefy 1-2 hour at 90 DEG C-105 DEG C, it is cooled to 60 DEG C-65 DEG C, it is 4-4.5 with acid for adjusting pH, the amount pressing 120u/g-200u/g starch again adds saccharifying enzyme, saccharifying 40-60 minute at 55 DEG C-65 DEG C, separate 15-20 minute with the centrifuge of at least 4000 revs/min after saccharifying, obtain the Dioscorea zingiberensis sugar liquid of centrifugal clear liquid.
Implement the Wine brewing yeast strain of the present invention and application thereof and cultural method, have the advantages that the fermentation of the novel Wine brewing yeast strain in present invention Dioscorea zingiberensis sugar liquid can tolerate pol and reaches 20Bx, can grow in the Dioscorea zingiberensis sugar liquid of different pols, breed, cell proliferation speed is fast, and sugar utilization is high-new, the direct defibrination even if cadmium yellow Rhizoma Zingiberis Recens does not add a little water, can ferment after liquefaction, saccharifying, inoculation, and fermentability is strong, fermenting speed is fast, only has 20-48 hour during general ferment；The bacterial strain reducing sugar utilization rate of the present invention is high, and fermentation liquid residual reducing sugar only has 0.1-0.3g/100ml, and remaining total sugar only has 0.5-1.0g/100ml, and the rate of getting alcohol, up to 45-50%, has reached cereal starch distillation yield level；The bacterial strain of the present invention can reach to make full use of Dioscorea zingiberensis, Rhizoma Dioscoreae starch or Dioscorea zingiberensis sugar liquid and produce the purpose of ethanol, improves Saponin yield simultaneously, improves the added value of Dioscorea zingiberensis, decreases the acid amount extracted used by diosgenin, decreases environmental pollution.
Detailed description of the invention
Below in conjunction with embodiment, the Wine brewing yeast strain of the present invention and application thereof and cultural method are described further:
Adopting different commercial yeast to inoculate in Dioscorea zingiberensis sugar liquid in an embodiment of the present invention, increase during along with ferment, living cells is fewer and feweri, and all dead after couple of days, fermentation cannot be carried out at all, and repeatedly fermentation test all terminates with failure.Owing to existing containing the material suppressing yeast growth in Dioscorea zingiberensis, general Alcohol Plant is usually used in alcohol fermentation commercial yeast (Angel, Dan Baoli etc.) and cannot be used for Dioscorea zingiberensis sugar liquid fermenting and producing ethanol.Cannot ferment the problem of Dioscorea zingiberensis sugar liquid to solve commercial yeast, to yeast through long-term mutation, raise and train, the process such as separation, finally obtain the novel saccharomyces cerevisiae new strains adapting to Dioscorea zingiberensis sugar liquid fermenting and producing ethanol, this bacterial strain can grow in the Dioscorea zingiberensis sugar liquid of different pols, propagation, cell proliferation speed is fast, and sugar utilization is high, can reach to make full use of Dioscorea. zingiberensis Wright Starch and produce the purpose of ethanol, simultaneously to improving Saponin yield, reduce environmental pollution and also there is important function.
The saccharomyces cerevisiae new strains of the present invention is through long-term mutation to yeast existing in prior art, raise and train, the bacterial strain that the processes such as separation obtain, this saccharomyces cerevisiae (Saccharomycescerevisaie), code name SDSW-HJ-1, its deposit number is CGMCCNo.9518, this bacterial strain on August 15th, 2014 in (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, China Committee for Culture Collection of Microorganisms's common micro-organisms center, Institute of Microorganism, Academia Sinica) preservation, Classification And Nomenclature is saccharomyces cerevisiae (Saccharomycescerevisaie).
Wine brewing yeast strain structure and physiological cells structure: 1. this bacterial strain belongs to saccharomyces cerevisiae, the same with general saccharomyces cerevisiae, there are the construction featuress such as cell wall, Cytoplasm, nucleus, mitochondrion, this bacterial strain bacterium colony is bigger and thick than cell, diameter about 2mm；
2. bacterium colony: bacterium colony smooth surface, is generally oval, circular or cylindrical, and its head is bigger than antibacterial, about 1～5 μ m 5～30 μm.Bacterium colony is moistening, thickness, it is easy to provoking, bacterium colony is homogeneous, and the color at positive and negative and edge, central part is homogeneous, is of different shades with culture medium, and bacterium colony is generally the light brown of different depth；
3. breathe: battalion's amphimicrobian life, the environment of aerobic carries out aerobic respiration, saccharide can be completely decomposed into CO2And H2O obtains substantial amounts of energy；Carry out anaerobic respiration under anoxic conditions, carbohydrate breakdown can be become C2H5OH (is commonly called as ethanol) and CO2, obtain a small amount of energy；It is in that to survive with Dioscorea zingiberensis for the converted mash of waste with the difference that general saccharomyces cerevisiae is maximum, can be fermented by the reducing sugar in mash and generate ethanol, and other commodity yeast then can not survived with Dioscorea zingiberensis for the converted mash of waste；
4. acidity: this bacterial strain can grow in the scope that pH value is 3.0～7.5, but optimum PH is 4.5～5.0；
5. temperature: this bacterial strain can be survived under 4 DEG C～46 DEG C environment, but optimum growth temperature is 28 DEG C～32 DEG C；
6. pol.This bacterial strain can at 24BxGrow under the environment of pol, but the suitableeest pol is 8Bx～14Bx。
Embodiment 1: the cultural method of Wine brewing yeast strain
1. the preparation of Dioscorea zingiberensis sugar liquid
Take Dioscorea zingiberensis 1000g to be cut into small pieces, add suitable quantity of water, it is crushed to particle diameter≤1mm screened stock with fiberizer or high speed disintegrator, add α-amylase by the amount of 8u/g starch, after liquefying 1 hour at 100 DEG C, be cooled to 62 DEG C, with breast acid for adjusting pH value to 4-4.5, the amount pressing 120u/g starch again adds α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme, saccharifying 50 minutes at 60 DEG C.Separate 18 minutes with 4000 revs/min of centrifuges after saccharifying is complete, obtain the Dioscorea zingiberensis sugar liquid of centrifugal clear liquid.
2. Wine brewing yeast strain culture medium prescription and growth preservation condition
Strain culturing based formulas: Dioscorea zingiberensis sugar liquid 500ml, MgSO40.015g, Co (NH2)20.175g, (NH4)2HPO40.125g, agar powder 10g, pH are natural.
Strain cultures is heated, agar powder injects in culture dish and test tube after dissolving, after 126 DEG C of autoclavings 30 minutes, cooling, makes flat board and slant tube, aseptically, by in inoculation to plating medium, cultivating 3.5 days at 30 DEG C, the bigger colony inoculation of picking is to slant tube, after cultivating then through 3.5 days, slant tube puts into cold compartment of refrigerator low-temperature preservation, preservation time 2-3 month.
3. yeast wine culture fluid formula and growth conditions
Yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 500ml, MgSO40.015g, Co (NH2)20.175g, (NH4)2HPO40.125g, pH are natural.
The bacterial strain preserved in slant tube is aseptically inoculated in the liquid tube equipped with 10ml yeast wine culture fluid, when 30 DEG C, after cultivating 14 hours, then is transferred in the liquid tube equipped with 10ml yeast wine culture fluid and continues cultivation 14 hours.Later by the continuous amplification culture of inoculum concentration 8-15%, spread cultivation step by step through little triangular flask, big triangular flask, Ka Shi tank etc., until required yeast wine amount.
4. amplification culture process wherein produces with the cultivation of little yeast wine:
Little yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 200L, MgSO46g, Co (NH2)270g, (NH4)2HPO450g, pH are natural.
Little yeast wine cultivation temperature 28 DEG C, incubation time 10 hours, per hour logical filtrated air 1 minute.
5. amplification culture process wherein produces with the cultivation of big yeast wine:
Big yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 2000L, MgSO420g, Co (NH2)2160g, (NH4)2HPO4140g, pH are natural.
Big yeast wine cultivation temperature 30 DEG C, incubation time 10 hours, per hour logical filtrated air 1 minute.
Embodiment 2: the application of Wine brewing yeast strain-utilize Dioscorea zingiberensis to prepare ethanol
By Dioscorea zingiberensis cleaning, remove impurity, pulverizing, add water and grind to form the slurry of granularity 2mm, wherein the solid content of slurry is 15%-30% by weight percentage；Add slurry into amylase to carry out enzymatic liquefaction and obtain starch fluid, temperature during enzymatic liquefaction controls at 85 DEG C-110 DEG C, the time of enzymatic liquefaction is 60min-120min, wherein diastatic vigor is 20000u/ml-90000u/ml, amylase can be α-amylase, beta amylase, α-1, at least one in 6 key debranching enzymes, namely can be α-amylase, beta amylase, α-1, any one in 6 key debranching enzymes, can also be α-amylase, beta amylase, α-1, the mixture of any two or three in 6 key debranching enzymes；Preferably by α-amylase, its enzymatic liquefaction best results；The starch fluid temperature of post liquefaction is down to 60 DEG C-65 DEG C, starch fluid adds saccharifying enzyme saccharifying 20min-60min and obtains converted mash, wherein the vigor of saccharifying enzyme is 80000u/ml-120000u/ml, and saccharifying enzyme can be α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme and/or beta-glucosidase；Take converted mash 20000L, MgSO46g, Co (NH2)2600g, (NH4)2HPO4400g, pH is natural, the amount of 10%-15% adds above-mentioned little yeast wine or big yeast wine ferments in mass ratio, cultivation temperature 30 DEG C-35 DEG C, incubation time is 20-48 hour, obtain fermenting-ripening wine with dregs, after solid-liquid separation, obtain fermentation liquid, fermentation liquid is distilled the ethanol of prepared concentration (V/V) more than 95%.Acquired ethanol and ethanol, it is possible to continue on for extracting remaining filtering residue administration Chinese yam Saponin after above-mentioned solid-liquid separation.
Embodiment 3: the Wine brewing yeast strain of the present invention and existing commodity culture propagation prepare the contrast experiment of ethanol
Contrast and experiment is referring to table 1, as can be seen from Table 1, the Wine brewing yeast strain of the application of the invention can utilize Dioscorea zingiberensis or its starch included, the preparation of Dioscorea zingiberensis sugar liquid to ferment excellent ethanol really, and solving general commercial yeast of the prior art cannot by the problem that Rhizoma Dioscoreae Starch Conversion is ethanol in Dioscorea zingiberensis.
Table 1:
Illustrate: the unit of reducing sugar and total sugar is g/100ml；Yeast wine used is to spread cultivation the Wine brewing yeast strain of the present invention by Dioscorea zingiberensis sugar liquid to obtain；Angel Yeast and Dan Baoli yeast by 2 ‰ i.e. 0.1g, by 50ml, 38 DEG C, the sucrose solution rehydration of 2% 20 minutes, are respectively inoculated in 500ml mash after activating 2 hours at 32 DEG C.
The explanation of following cultural method for other embodiments to Wine brewing yeast strain and application.
Embodiment 4: the cultural method of Wine brewing yeast strain
1. the preparation of Dioscorea zingiberensis sugar liquid
Take Dioscorea zingiberensis 1000g to be cut into small pieces, add suitable quantity of water, it is crushed to particle diameter≤1mm screened stock with fiberizer or high speed disintegrator, add beta amylase by the amount of 12u/g starch, after liquefying 2 hours at 90 DEG C, be cooled to 60 DEG C, with breast acid for adjusting pH value to 4-4.5, the amount pressing 160u/g starch again adds α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme, saccharifying 40 minutes at 55 DEG C.Separate 15 minutes with 6000 revs/min of centrifuges after saccharifying is complete, obtain the Dioscorea zingiberensis sugar liquid of centrifugal clear liquid.
2. Wine brewing yeast strain culture medium prescription and growth preservation condition
Strain culturing based formulas: Dioscorea zingiberensis sugar liquid 450ml, MgSO40.01g, Co (NH2)20.17g, (NH4)2HPO40.12g, agar powder 9g, pH are natural.
Strain cultures is heated, agar powder injects in culture dish and test tube after dissolving, after 120 DEG C of autoclavings 70 minutes, cooling, makes flat board and slant tube, aseptically, by in inoculation to plating medium, cultivating 4 days at 28 DEG C, the bigger colony inoculation of picking is to slant tube, after cultivating then through 4 days, slant tube puts into cold compartment of refrigerator low-temperature preservation, preservation time 2-3 month.
3. yeast wine culture fluid formula and growth conditions
Yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 450ml, MgSO40.01g, Co (NH2)20.17g, (NH4)2HPO40.12g, pH are natural.
The bacterial strain preserved in slant tube is aseptically inoculated in the liquid tube equipped with 10ml yeast wine culture fluid, when 32 DEG C, after cultivating 16 hours, then is transferred in the liquid tube equipped with 10ml yeast wine culture fluid and continues cultivation 16 hours.Later by the continuous amplification culture of inoculum concentration 8-15%, spread cultivation step by step through little triangular flask, big triangular flask, Ka Shi tank etc., until required yeast wine amount.
4. amplification culture process wherein produces with the cultivation of little yeast wine:
Little yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 150L, MgSO45g, Co (NH2)260g, (NH4)2HPO440g, pH are natural.
Little yeast wine cultivation temperature 30 DEG C, incubation time 12 hours, per hour logical filtrated air 3 minutes.
5. amplification culture process wherein produces with the cultivation of big yeast wine:
Big yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 1500L, MgSO415g, Co (NH2)2120g, (NH4)2HPO4100g, pH are natural.
Big yeast wine cultivation temperature 32 DEG C, incubation time 8 hours, per hour logical filtrated air 3 minutes.
Embodiment 5: the cultural method of Wine brewing yeast strain
1. the preparation of Dioscorea zingiberensis sugar liquid
Take Dioscorea zingiberensis 1000g to be cut into small pieces, add suitable quantity of water, it is crushed to particle diameter≤1mm screened stock with fiberizer or high speed disintegrator, add beta amylase by the amount of 16u/g starch, after liquefying 1.5 hours at 105 DEG C, be cooled to 65 DEG C, with breast acid for adjusting pH value to 4-4.5, the amount pressing 200u/g starch again adds α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme, saccharifying 60 minutes at 65 DEG C.Separate 20 minutes with 5000 revs/min of centrifuges after saccharifying is complete, obtain the Dioscorea zingiberensis sugar liquid of centrifugal clear liquid.
2. Wine brewing yeast strain culture medium prescription and growth preservation condition
Strain culturing based formulas: Dioscorea zingiberensis sugar liquid 550ml, MgSO40.02g, Co (NH2)20.18g, (NH4)2HPO40.13g, agar powder 11g, pH are natural.
Strain cultures is heated, agar powder injects in culture dish and test tube after dissolving, after 130 DEG C of autoclavings 120 minutes, cooling, makes flat board and slant tube, aseptically, by in inoculation to plating medium, cultivating 3 days at 32 DEG C, the bigger colony inoculation of picking is to slant tube, after cultivating then through 3 days, slant tube puts into cold compartment of refrigerator low-temperature preservation, preservation time 2-3 month.
3. yeast wine culture fluid formula and growth conditions
Yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 550ml, MgSO40.02g, Co (NH2)20.18g, (NH4)2HPO40.13g, pH are natural.
The bacterial strain preserved in slant tube is aseptically inoculated in the liquid tube equipped with 10ml yeast wine culture fluid, when 31 DEG C, after cultivating 10 hours, then is transferred in the liquid tube equipped with 10ml yeast wine culture fluid and continues cultivation 10 hours.Later by the continuous amplification culture of inoculum concentration 8-15%, spread cultivation step by step through little triangular flask, big triangular flask, Ka Shi tank etc., until required yeast wine amount.
4. amplification culture process wherein produces with the cultivation of little yeast wine:
Little yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 240L, MgSO47g, Co (NH2)265g, (NH4)2HPO460g, pH are natural.
Little yeast wine cultivation temperature 29 DEG C, incubation time 8 hours, per hour logical filtrated air 5 minutes.
5. amplification culture process wherein produces with the cultivation of big yeast wine:
Big yeast wine culture fluid formula: Dioscorea zingiberensis sugar liquid 2400L, MgSO425g, Co (NH2)2200g, (NH4)2HPO4180g, pH are natural.
Big yeast wine cultivation temperature 31 DEG C, incubation time 12 hours, per hour logical filtrated air 5 minutes.
Embodiment 6: the application of Wine brewing yeast strain-utilize Rhizoma Dioscoreae starch to prepare ethanol
Adding saccharifying enzyme saccharifying 20min-60min in Rhizoma Dioscoreae starch and obtain converted mash, wherein the vigor of saccharifying enzyme is 80000u/ml-120000u/ml, and saccharifying enzyme can be α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme and/or beta-glucosidase；Take converted mash 20000L, MgSO46g, Co (NH2)2600g, (NH4)2HPO4400g, pH are natural, add above-mentioned little yeast wine or big yeast wine ferments, cultivation temperature 30 DEG C-35 DEG C, incubation time is 20-48 hour, obtains fermenting-ripening wine with dregs, obtain fermentation liquid after solid-liquid separation, fermentation liquid is distilled the ethanol of prepared concentration (V/V) more than 95%.
Embodiment 7: the application of Wine brewing yeast strain-utilize Dioscorea zingiberensis sugar liquid to prepare ethanol
Take Dioscorea zingiberensis sugar liquid 20000L, MgSO46g, Co (NH2)2600g, (NH4)2HPO4400g, pH are natural, add above-mentioned little yeast wine or big yeast wine ferments, cultivation temperature 30 DEG C-35 DEG C, incubation time is 20-48 hour, obtains fermenting-ripening wine with dregs, obtain fermentation liquid after solid-liquid separation, fermentation liquid is distilled the ethanol of prepared concentration (V/V) more than 95%.
Embodiment 8: the fermenting-ripening wine with dregs that different Dioscorea zingiberensis utilize Wine brewing yeast strain to produce
Referring to table 2, it can be seen that different Dioscorea zingiberensis can be prepared stable fermenting-ripening wine with dregs by the yeast wine that the Wine brewing yeast strain of the present invention is cultivated.
Table 2:
It should be appreciated that for those of ordinary skills, it is possible to being improved according to the above description or convert, all these improve or convert within the protection domain that all should belong to claims of the present invention.
Claims (10)
1. a Wine brewing yeast strain (Saccharomycescerevisaie), its deposit number is CGMCCNo.9518.
2. the Wine brewing yeast strain described in claim 1 prepares the application in ethanol in production.
3. Wine brewing yeast strain according to claim 2 is prepared in the application in ethanol in production, it is characterized in that, described Wine brewing yeast strain is for preparing ethanol by Dioscorea zingiberensis production, for preparing ethanol or for preparing ethanol by Dioscorea zingiberensis sugar liquid fermenting and producing by Rhizoma Dioscoreae Starch Production.
4. the cultural method of the Wine brewing yeast strain described in a claim 1, it is characterised in that comprise the steps:
S1, bacterial strain cultivation preserve: being heated by strain cultures, agar powder injects in culture dish and test tube after dissolving, cooling after 120 DEG C of-130 DEG C of autoclavings 30-120 minute, makes flat board and slant tube；Aseptically, in inoculation to plating medium, cultivating 3-4 days at 28-32 DEG C, the bigger colony inoculation of picking, to slant tube, was cultivated then through 3-4 days, and slant tube puts into cold compartment of refrigerator cryopreservation；
S2, bacterial strain activation culture obtain yeast wine: be aseptically inoculated in the liquid tube equipped with 10ml yeast wine culture fluid by the bacterial strain preserved in step S1, when 30-32 DEG C, after cultivating 10-16 hour, then it is transferred in the liquid tube equipped with 10ml yeast wine culture fluid and continues cultivation 10-16 hour.
5. cultural method according to claim 4, it is characterised in that in step S1, the formula of strain cultures is: Dioscorea zingiberensis sugar liquid 450ml-550ml, MgSO40.01g-0.02g, Co (NH2)20.17g-0.18g, (NH4)2HPO40.12g-0.13g, agar powder is 9-11g.
6. cultural method according to claim 4, it is characterised in that the formula of the yeast wine culture fluid in step S2 is: Dioscorea zingiberensis sugar liquid 450ml-550ml, MgSO40.01g-0.02g, Co (NH2)20.17g-0.18g, (NH4)2HPO40.12g-0.13g。
7. cultural method according to claim 4, it is characterised in that also include step S3 upon step s 2, the yeast wine obtained in step S2 for seed by inoculum concentration 8%-15% constantly amplification culture step by step, until required yeast wine amount.
8. cultural method according to claim 7, it is characterised in that in step s3 in the process of yeast wine amplification culture step by step, includes the step A that the little yeast wine for producing is cultivated, and its medium and small yeast wine culture fluid formula is Dioscorea zingiberensis sugar liquid 150L-240L, MgSO45g-7g, Co (NH2)260g-70g, (NH4)2HPO440g-60g；Condition of culture: cultivation temperature 28 DEG C-30 DEG C, incubation time 8-12 hour, per hour logical filtrated air 1-5 minute.
9. cultural method according to claim 7, it is characterised in that in step s3 in the process of yeast wine amplification culture step by step, includes the step A that the big yeast wine for producing is cultivated, and wherein big yeast wine culture fluid formula is Dioscorea zingiberensis sugar liquid 1500L-2400L, MgSO415g-25g, Co (NH2)2120g-200g, (NH4)2HPO4100g-180g；Condition of culture: cultivation temperature 30 DEG C-32 DEG C, incubation time 8-12 hour, per hour logical filtrated air 1-5 minute.
10. the cultural method according to claim 5 or 6 or 8 or 9, it is characterized in that, described Dioscorea zingiberensis sugar liquid is prepared as follows: take Dioscorea zingiberensis by pulverizing and/or grind to form the screened stock of particle diameter≤1mm, amylase is added by the amount of 8u/g-16u/g starch, liquefy 1-2 hour at 90 DEG C-105 DEG C, it is cooled to 60 DEG C-65 DEG C, it is 4-4.5 with acid for adjusting pH, the amount pressing 120u/g-200u/g starch again adds saccharifying enzyme, saccharifying 40-60 minute at 55 DEG C-65 DEG C, separate 15-20 minute with the centrifuge of at least 4000 revs/min after saccharifying, obtain the Dioscorea zingiberensis sugar liquid of centrifugal clear liquid.
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