CN103525711A - Three-stage composite fermentation microbial inoculum for Sichuan-Tibet Plateau ice wine and preparation method thereof - Google Patents
Three-stage composite fermentation microbial inoculum for Sichuan-Tibet Plateau ice wine and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of a three-stage composite fermentation microbial inoculum for Sichuan-Tibet Plateau ice wine. Pichia manshurica (culture collection number: CGMCC 6867), Pichia anomala (culture collection number: CGMCC 6866), Saccharomyces cerevisiae and Saccharomyces chevalieri are used as main fermentation microbes to prepare the earlier stage aging-medium stage quick start-later stage fermentation three-stage composite fermentation microbial inoculum for Sichuan-Tibet Plateau ice wine by a modern biological technique and a microcapsule preparation manufacturing technique. The invention provides a core technique for ice standardized production, fills up the domestic production blank, can have an active promotion action on development of high-end products and adaptation to the market consumption demands in Chinese grape and wine industry, and conforms to the demands for actual local feature industries.
Description
Technical field
The present invention relates to complex ferment preparation, in particular three grades of complex ferment microbial inoculum preparation methods of plateau, a kind of Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage.
Background technology
The discovery in ice-wine producing region, Tibetan area, plateau, China river and the construction of ice-wine industry project, the extremely strong irreplaceable not reproducible natural resource advantages of region will be converted into market competition advantage, facilitate and there is the Industry with Local Characteristics of international competitiveness and the formation of mainstay industry.Ice-wine is as the noble in grape wine, and aroma is naturally sweet pure and sweet, and clear limpid, mouthfeel is meaningful, and quality is superior.Meanwhile, in ice-wine, nutritive ingredient is abundant, has almost comprised the necessary tens of kinds of enzymes of human body and amino acid, has the physiological functions such as beauty treatment, beauty treatment, health care, is rare superior vintage wine.Ice-wine has now become the representative of high-end product on world's Wine Market.But owing to being subject to the restriction of scientific knowledge, ice-wine is produced in family workshop type and nucleus estate, utilizes spontaneous fermentation to brewage ice-wine always, industry development speed is very slow.It produces nature inoculation, the spontaneous fermentation stage of still maintaining, and within it is sold and is only limited to territory, county coverage, therefore, the research and development that traditional ice grape industry is carried out to technology upgrading and Key Techniques in Standardized Production are extremely urgent.And the key of ice-wine standardized production is the exploitation of ice-wine specific complex fermenting agent, the starters such as fruit wine that sell in market easily make the original local flavor desalination of ice-wine, lose local characteristic and unique local flavor, can not meet the ice-wine fermentation of Tibetan area, plateau, Sichuan.
Summary of the invention
Technical problem to be solved by this invention is to provide three grades of complex ferment microbial inoculums of plateau, a kind of Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage and preparation method thereof for the deficiencies in the prior art.
Technical scheme of the present invention is as follows:
A preparation method for three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine, comprises the following steps:
(1) saccharomycetic enlarged culturing: the yeast Pichia manshurica of deposit number: CGMCC6867, the Pichia anomala of deposit number: CGMCC6866 are inoculated in substratum, then be placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension; By the mixed strains centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 1, centrifugal concentrating thing standby;
(2) enlarged culturing of S. cervisiae: S. cervisiae Saccharomyces cerevisiae and Xue watt yeast Saccharomyces chevalieri are inoculated in substratum, are then placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension;
(3) will go ice grape pomace after seed after high-temperature sterilization, by the 5%-13% of pomace weight, add described centrifugal concentrating thing No. 1 and No. 2, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:2～5:4, put into 20-28 ℃ of fermenting case quick fermentation 10-18h, in-45～-30 ℃ of Ultralow Temperature Freezers after freezing 10-16h or adopt after carbonic acid gas dry ice moment quick freezing, vacuum freeze drier is put in taking-up, in condenser temperature-45～-30 ℃, material bin temperature is under-30～-35 ℃ of conditions, vacuum tightness is to take out after the dry 24h of 0.90-0.95MPa condition, after high speed disintegrator is pulverized, obtain ice-wine early stage No. 1, stuffed fermenting agent standby,
(4) mid-term, quick fermentation started bacterial preparation process: get described bacterial classification centrifugal concentrating thing and be mixed in proportion for No. 1 and No. 2, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:5～4:7, with 7%～10% (W/V that accounts for No. 1 and No. 2 mixture amount of centrifugal concentrating thing, following per-cent is all W/V) water-soluble chitosan, 3% whey protein concentrate, the embedding medium that 10% porous-starch mixes; Or with 20% the chitosan that accounts for No. 1 and No. 2 mixture amount of centrifugal concentrating thing, 8% whey protein concentrate, the embedding medium that 10% glucose mixes, obtaining ice-wine, to start fast No. 2, fermenting agent mid-term standby;
(5) later stage fermentation bacterial preparation process: get described bacterial classification centrifugal concentrating thing No. 1 and No. 2 porous-starch of totally 15～18g and 3～5g to join pH value be that in 5.0～6.5 the aqueous solution, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 4:1～2:1; Vibration is stirred 0.5～1.5h after adding the sodium alginate of 6～8g after 1.5～2.5h, then by the 2.5%CaCl of 5～10mL
2solution dropwise adds, after solidifying 25～35min, filter, then added in the ethanolic soln that contains 3～7% the molten corn protein concentrate of alcohol and the molten wheat protein of 4%～6% alcohol, high speed 1200r/min stirs 1.0～1.5h, gained mixture is put into spraying plant, the inlet temperature of device is 83～87 ℃, and temperature out is 61～67 ℃, and No. 3, gained later stage fermentation microbial inoculum is standby;
By starting fast No. 2, fermenting agent and later stage fermentation microbial inoculum in described early stage No. 1, stuffed fermenting agent, mid-term, according to 1:3:1.5～1:5:3, obtain weight ratio No. 3 and mix, or by No. 1 independent vacuum small packages of stuffed fermenting agent in early stage, start fast No. 3 weight ratios according to 3:2～5:4 of No. 2, fermenting agent and later stage fermentation microbial inoculum mid-term and mix final vacuum small packages, and two small packagess are become to three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage through the large packing of vacuum.
Described preparation method, in described step (1), described medium component is: 1.8%～2.3% peptone, 1.9%～2.2% glucose, 0.9%～1.1% yeast powder, pH6.8～7.2.
Described preparation method, in described step (2), described substratum comprises 4.5～5.5 ° of B é wort 1.0L, agar 14.5～15.5g, natural pH.By the bacterial classification centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 2, centrifugal concentrating thing standby.
The present invention is with Pichia manshurica (culture presevation numbering: CGMCC6867), Pichia anomala (abnormal pichia spp, culture presevation numbering: CGMCC6866), Saccharomyces cerevisiae (yeast saccharomyces cerevisiae, purchase is from Chinese industrial microbial strains preservation administrative center, CICC1450) and Saccharomyces chevalieri (Xue's watt yeast, purchase is from Chinese industrial microbial strains preservation administrative center, CICC1424) be main zymophyte, adopt modern biotechnology and microcapsule formulation technology of preparing, make three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage, for ice-wine standardized production provides core technology, fill up domestic production blank, the development of Zhe Jiangdui China grape and Grape Wine Industry mid-to high-end product and adaptation market comsupton demand play positive promoter action, met real Industry with Local Characteristics development need.
Principal feature of the present invention: (1) adopts three grades of complex ferment microbial inoculums of start fast-later stage fermentation in stuffed-mid-term in early stage to make plateau, Sichuan-Tibet ice-wine can realize suitability for industrialized production, fills up domestic production blank.(2) main bacteria seed that these three grades of complex ferment microbial inoculums are used is the major function bacterium of separating from traditional ice-wine fermenting process, adopt these three grades of composite zymocytes to carry out the fermentation of ice-wine, kept local flavor and the feature of plateau, traditional Sichuan-Tibet ice-wine.(3) adopt the material of different performance as the embedding wall material of composite fungus agent, thereby realize the three grade fermemtation of start fast-later stage fermentation in stuffed-mid-term in early stage.(4) adopt these three grades of complex ferment microbial inoculums to make plateau, Sichuan-Tibet ice-wine fermentation time shorten 25%-35%, production efficiency improves 80%-100%, fills up domestic production blank.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
From Aba Prefecture Li County, pluck the processing of squeezing the juice of ripe Vidal Blanc ice grape, with the red substratum of tiger, from Sucus Vitis viniferae, filter out the vigorous and obvious 2 strain bacterium of feature of growth, by plate streaking, isolate single bacterium colony of this strain bacterium, picking list bacterium colony is preserved bacterial strain with glycerine after cultivating in liquid PDA nutrient solution, be placed on-20 ℃ standby.Get yeast strain streak inoculation that separation obtains in WL substratum, cultivate 5d for 28 ℃, the colony characteristics such as whether color, form, the surface of observing bacterium colony be smooth, whether edge is neat.Meanwhile, examine under a microscope the cellular form of yeast.With this, yeast strain being separated to is carried out to preliminary typoiogical classification.On the basis of typoiogical classification, select 2 saccharomycetes (XHU410 and XHU411) of different shape to adopt 18SrDNA sequence analysis to carry out Molecular Identification to isolated bacterial classification, after sequencing result and database NCBI Blast compare, by Clustal X1.8, will represent that in sequence and database, institute's calling sequence is compared completely again, determine that this two Yeasts is respectively Pichia manshurica (XHU411) and Pichia anomala (XHU410), and on November 22nd, 2012, deliver (address: preservation Datun Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica), China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation information is respectively: XHU411, Classification And Nomenclature Pichia manshurica, deposit number: CGMCC6867, XHU410, Classification And Nomenclature: abnormal pichia spp Pichia anomala, deposit number: CGMCC6866).
Saccharomycetic enlarged culturing: by above-mentioned two saccharomycete Pichia manshurica (culture presevation numbering: CGMCC6867), Pichia anomala (culture presevation numbering: CGMCC6866) be inoculated in substratum, then be placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension; Described medium component is; 1.8%～2.3% peptone, 1.9%～2.2% glucose, 0.9%～1.1% yeast powder, pH6.8～7.2.By the mixed strains centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 1, centrifugal concentrating thing standby.
The enlarged culturing of S. cervisiae: by Saccharomyces cerevisiae (S. cervisiae, purchase is from Chinese industrial microbial strains preservation administrative center, CICC1450) and Saccharomyces chevalieri (Xue's watt yeast, purchase is from Chinese industrial microbial strains preservation administrative center, CICC1424) be inoculated in substratum, then be placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension; Described substratum comprises 4.5～5.5 ° of B é wort 1.0L, agar 14.5～15.5g, natural pH.By the bacterial classification centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 2, centrifugal concentrating thing standby.
In three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage, early stage stuffed fermenting agent preparation method: will go ice grape pomace after seed after high-temperature sterilization, No. 1 and No. 2 (No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:2～5:4) by the 5%-13% of pomace weight, to add above-mentioned centrifugal concentrating thing, put into 20-28 ℃ of fermenting case quick fermentation 10-18h, in-45～-30 ℃ of Ultralow Temperature Freezers after freezing 10-16h or adopt after carbonic acid gas dry ice moment quick freezing, vacuum freeze drier is put in taking-up, in condenser temperature-45～-30 ℃, material bin temperature is under-30～-35 ℃ of conditions, vacuum tightness is to take out after the dry 24h of 0.90-0.95MPa condition, after high speed disintegrator is pulverized, obtain ice-wine early stage No. 1, stuffed fermenting agent standby.
Mid-term, quick fermentation started bacterial preparation process: get above-mentioned bacterial classification centrifugal concentrating thing and be mixed in proportion that (No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:5～4:7) for No. 1 and No. 2, with 7%～10% (W/V that accounts for No. 1 and No. 2 mixture amount of centrifugal concentrating thing, following per-cent is all W/V) water-soluble chitosan, 3% whey protein concentrate, the embedding medium that 10% porous-starch mixes; Or with 20% the chitosan that accounts for composite yeast bacteria suspension total amount, 8% whey protein concentrate, the embedding medium that 10% glucose mixes, obtaining ice-wine, to start fast No. 2, fermenting agent mid-term standby.
Later stage fermentation bacterial preparation process: No. 1 and No. 2 (No. 1, centrifugal concentrating thing and No. 2 part by weight are: 4:1～2:1) 15～18g to get above-mentioned bacterial classification centrifugal concentrating thing, it is in 5.0～6.5 the aqueous solution that the porous-starch of 3～5g joins pH value, vibration is stirred 0.5～1.5h after adding the sodium alginate of 6～8g after 1.5～2.5h, then by the 2.5%CaCl of 5～10mL
2solution dropwise adds, after solidifying 25～35min, filter, then added in the ethanolic soln that contains 3～7% the alcohol molten protein concentrate of molten corn alcohol and the molten wheat protein of 4%～6% alcohol (ethanol content is 15～18%), high speed 1200r/min stirs 1.0～1.5h, adopt low temperature spray drying method, above-mentioned gained mixture is put into spraying plant, the inlet temperature of device is 83～87 ℃, temperature out is 61～67 ℃, and No. 3, gained later stage fermentation microbial inoculum is standby.
Three grades of complex ferment microbial inoculums of plateau, a kind of Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage and preparation method thereof:
By above-mentioned early stage stuffed fermenting agent, mid-term starts fast fermenting agent and later stage fermentation microbial inoculum mixes according to 1:3:1.5～1:5:3 (weight), or by the independent vacuum small packages of stuffed fermenting agent in early stage, start fast fermenting agent and later stage fermentation microbial inoculum mid-term and mix final vacuum small packages according to 3:2～5:4 (weight), and the two become to three grades of complex ferment microbial inoculums of plateau, a kind of Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage and preparation method thereof through the large packing of vacuum.
The material of employing different performance of the present invention is as the embedding wall material of composite fungus agent, the main bacteria seed of using is the major function bacterium of separating from traditional ice-wine fermenting process, adopt advanced modern technologies, three grades of complex ferment microbial inoculums of start fast-later stage fermentation in stuffed-mid-term in early stage have been prepared, thereby realize the three grade fermemtation of start fast-later stage fermentation in stuffed-mid-term in early stage, and local flavor and the feature of plateau, traditional Sichuan-Tibet ice-wine have been kept, make ice-wine fermentation time shorten 25%-35%, production efficiency improves 80%-100%, this composite fungus agent makes plateau, Sichuan-Tibet ice-wine can realize suitability for industrialized production, fill up domestic production blank.
Application example:
The present invention provide on the other hand a kind of early stage stuffed-three grades of start fast mid-term-later stage fermentations complex ferment microbial inoculum produces the making method of ice grape.Concrete technology flow process is divided into four-stage, the pre-treatment of (1) ice grape material; (2) add complex ferment microbial inoculum; (3) start three grade fermemtation; (4) post-treatment is processed.
Freezing grape → harvesting, sorting → squeezing → original grape juice add 5-7% complex ferment microbial inoculum → stuffed → clarification → startup fermentation → freezing → later stage fermentation → filtration sterilization → filling → finished product ice wine.
1) pluck sorting
Delay grape collection period to severe winter, allow grape through the freezing and process of thawing several times.Detect sugar degree and total acid content in Sucus Vitis viniferae, when Sucus Vitis viniferae sugar degree is about 320-360g/L (with glucose meter), when total acid is about 8.0-12.0g/L (in tartrate), pluck.Pluck sorting ice grape and carry out at-7 ℃-9 ℃, harvesting process by hand careful is plucked, and selects without sick green grass or young crops, the squeezing immediately of rotten fruit.
2) squeezing
In expressing process, adopt graded temperature-control Screw Presser group to squeeze, pressing temperature is strict controlled at-7 ℃～-9 ℃.Meanwhile, squeezing juice is pressed 80mg/L and is calculated interpolation sulfurous acid.
3) three grade fermemtation
3.1 microbial inoculums add
In the ice original grape juice having squeezed, add 5-7wt% complex ferment microbial inoculum, start stuffed process in early stage.
3.2 three grade fermemtation
Adopt three grade low-temp fermentation modes to ferment, the stuffed fermentation of one-level: leavening temperature is 7 ℃-8 ℃, fermentation time is 4-6 days, in the process of sinking at the grape pomace of pulverizing (can ferment with pericarp), the fermenting agent of the stuffed special use in early stage in the fermenting agent adding, can in the sedimentary sinking processes such as pericarp, start fermentation, and stir once every 2-3h, make sinking after its floating, repeatedly carry out stuffed, after 4-6 days, start the second order fermentation stage: leavening temperature is 12 ℃-15 ℃, fermentation time is 30-35 days, because the embedding wall material of the startup fermenting agent in mid-term is owing to dissolving gradually under the effect of water in Sucus Vitis viniferae, and final disintegration, discharge special bacteria agent, the fermentation in mid-term can be started fast, prolongation along with fermentation time, yeast viable count contained in fermented liquid reduces gradually, the alcoholic strength of wine body raises gradually simultaneously, when alcoholic strength reaches 8% when above, along with alcoholic strength raises gradually, the molten wheat protein of wall material Zeinfilm and alcohol is swelling cause final disintegration gradually, then being embedded in fermented yeast bacterium discharges gradually, start the three grade fermemtation stage: leavening temperature is 10 ℃-12 ℃, fermentation time is 10-15 days, wine body is carried out to the fermentation of later stage flavouring etc.Detect in wine the physical and chemical indexs such as alcoholic strength, total reducing sugar, sugar-free extract, volatile acid, will reach the subsequent disposal of carrying out that GB GB/T25504-2010 requires.
4) post-treatment is processed
Fermented wine, after several months ageing, is clarified naturally, and clarifying process temperature is no more than 8 ℃, adjusts free S0 simultaneously
2to 40～50mg/L.Then through freezing, filtration sterilization, sterile filling, make ice wine.
The present invention is by adopting plateau, the enlarged culturing Sichuan-Tibet ice-wine special-purpose yeast that ferments, select chitosan, the whey-protein embedding that combines with glucose, or employing beta-cyclodextrin, chitosan, whey protein concentrate, the porous-starch embedding that combines, has developed the composite fungus agent that is exclusively used in the ice-wine fermentation of plateau, Sichuan-Tibet.And experimental result shows, composite fungus agent of the present invention is produced for the high field ice grape fermentation in Sichuan-Tibet, the fermenting speed of ice-wine is obviously accelerated, shortened the fermentation time of ice-wine, the ice-wine of fermentation has good color and luster and pleasant fragrance, has improved the commercial value of ice-wine.And the preparation method of plateau, a kind of Sichuan-Tibet provided by the present invention ice-wine high-fidelity complex ferment microbial inoculum, processing step is simple, and material therefor cost is low, is suitable for suitability for industrialized production, and range of application is wide.
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (4)
1. a preparation method for three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine, is characterized in that, comprises the following steps:
(1) saccharomycetic enlarged culturing: the yeast Pichia manshurica of deposit number: CGMCC6867, the Pichia anomala of deposit number: CGMCC6866 are inoculated in substratum, then be placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension; By the mixed strains centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 1, centrifugal concentrating thing standby;
(2) enlarged culturing of S. cervisiae: S. cervisiae Saccharomyces cerevisiae and Xue watt yeast Saccharomyces chevalieri are inoculated in substratum, are then placed in the shaking table of 25 ℃ and cultivate 24h, to viable count be 10
8cfu/mL, makes mulriple yeasts bacteria suspension;
(3) will go ice grape pomace after seed after high-temperature sterilization, by the 5%-13% of pomace weight, add described centrifugal concentrating thing No. 1 and No. 2, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:2～5:4, put into 20-28 ℃ of fermenting case quick fermentation 10-18h, in-45～-30 ℃ of Ultralow Temperature Freezers after freezing 10-16h or adopt after carbonic acid gas dry ice moment quick freezing, vacuum freeze drier is put in taking-up, in condenser temperature-45～-30 ℃, material bin temperature is under-30～-35 ℃ of conditions, vacuum tightness is to take out after the dry 24h of 0.90-0.95MPa condition, after high speed disintegrator is pulverized, obtain ice-wine early stage No. 1, stuffed fermenting agent standby,
(4) mid-term, quick fermentation started bacterial preparation process: get described bacterial classification centrifugal concentrating thing and be mixed in proportion for No. 1 and No. 2, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 3:5～4:7, with 7%～10% (W/V that accounts for No. 1 and No. 2 mixture amount of centrifugal concentrating thing, following per-cent is all W/V) water-soluble chitosan, 3% whey protein concentrate, the embedding medium that 10% porous-starch mixes; Or with 20% the chitosan that accounts for No. 1 and No. 2 mixture amount of centrifugal concentrating thing, 8% whey protein concentrate, the embedding medium that 10% glucose mixes, obtaining ice-wine, to start fast No. 2, fermenting agent mid-term standby;
(5) later stage fermentation bacterial preparation process: get described bacterial classification centrifugal concentrating thing No. 1 and No. 2 porous-starch of totally 15～18g and 3～5g to join pH value be that in 5.0～6.5 the aqueous solution, No. 1, centrifugal concentrating thing and No. 2 part by weight are: 4:1～2:1; Vibration is stirred 0.5～1.5h after adding the sodium alginate of 6～8g after 1.5～2.5h, then by the 2.5%CaCl of 5～10mL
2solution dropwise adds, after solidifying 25～35min, filter, then added in the ethanolic soln that contains 3～7% the molten corn protein concentrate of alcohol and the molten wheat protein of 4%～6% alcohol, high speed 1200r/min stirs 1.0～1.5h, gained mixture is put into spraying plant, the inlet temperature of device is 83～87 ℃, and temperature out is 61～67 ℃, and No. 3, gained later stage fermentation microbial inoculum is standby;
By starting fast No. 2, fermenting agent and later stage fermentation microbial inoculum in described early stage No. 1, stuffed fermenting agent, mid-term, according to 1:3:1.5～1:5:3, obtain weight ratio No. 3 and mix, or by No. 1 independent vacuum small packages of stuffed fermenting agent in early stage, start fast No. 3 weight ratios according to 3:2～5:4 of No. 2, fermenting agent and later stage fermentation microbial inoculum mid-term and mix final vacuum small packages, and two small packagess are become to three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine start fast-later stage fermentation in stuffed-mid-term in early stage through the large packing of vacuum.
2. preparation method according to claim 1, is characterized in that, in described step (1), described medium component is: 1.8%～2.3% peptone, 1.9%～2.2% glucose, 0.9%～1.1% yeast powder, pH6.8～7.2.
3. preparation method according to claim 1, is characterized in that, in described step (2), described substratum comprises 4.5～5.5 ° of B é wort 1.0L, agar 14.5～15.5g, natural pH.By the bacterial classification centrifugal concentrating of enlarged culturing, centrifugal rotational speed is 4000r/min, and the time is 10min, obtains No. 2, centrifugal concentrating thing standby.
4. three grades of complex ferment microbial inoculums of plateau, Sichuan-Tibet ice-wine that prepared by preparation method according to claim 1 and 2.
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| CN106434086A (en) * | 2016-08-31 | 2017-02-22 | 朱新洁 | Production process of wine |
| CN107287073A (en) * | 2017-07-06 | 2017-10-24 | 泸州向林老窖股份有限公司 | A kind of compound Koji and its production technology |
| CN108251411A (en) * | 2018-02-13 | 2018-07-06 | 南阳师范学院 | A kind of method that grape wine fixed yeast is prepared using oak chip |
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| CN102443507B (en) * | 2011-12-27 | 2016-01-13 | 国家纳米技术与工程研究院 | A kind of production technique of preparing rose wine with composite bio-enzyme |
| CN102965291B (en) * | 2012-11-29 | 2015-04-22 | 天津大学 | Saccharomyces cerevisiae strain and application of strain in production of ethanol by co-fermentation of glucose and xylose |
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| CN106434086A (en) * | 2016-08-31 | 2017-02-22 | 朱新洁 | Production process of wine |
| CN107287073A (en) * | 2017-07-06 | 2017-10-24 | 泸州向林老窖股份有限公司 | A kind of compound Koji and its production technology |
| CN108251411A (en) * | 2018-02-13 | 2018-07-06 | 南阳师范学院 | A kind of method that grape wine fixed yeast is prepared using oak chip |
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| CN104120087A (en) | 2014-10-29 |
| CN104120087B (en) | 2017-03-15 |
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