CN107446860B - It is a kind of produce Tetramethylpyrazine bacillus licheniformis and its application - Google Patents
It is a kind of produce Tetramethylpyrazine bacillus licheniformis and its application Download PDFInfo
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Abstract
The invention discloses a kind of bacillus licheniformis for producing Tetramethylpyrazine and its applications, wherein the classification naming of the bacillus licheniformis is bacillus licheniformis GJBL055 (Bacillus licheniformis GJBL055), it has been preserved in China typical culture collection center, the deposit date is on April 24th, 2017, deposit number is CCTCC NO:M 2017204, and preservation address is Wuhan, China Wuhan University.Use medium temperature shelf training mode, bacillus licheniformis of the invention is linked into plate yeast, pass through the building of preculture, shelf song spatial model to functional microorganism, four stages of area room temperature regulate and control, the yeast manufacturing process for forming high yield Tetramethylpyrazine under a kind of medium temperature condition, makes Tetramethylpyrazine content in yeast be increased to 400mg/kg.
Description
Technical field
The present invention relates to a kind of bacillus licheniformis for producing Tetramethylpyrazine and its applications, belong to fermentation engineering neck
Domain.
Background technique
" bone that song is wine ", yeast plays the role of vital in liquor production.It prepares purpose are as follows: multi-functional bacterium
The formation of system, multi-enzyme system and a variety of bouquet principles matter.In yeast production technology, bacterium and saccharomycete ferment first, make song
Heart temperature increases, and the surface layer for forming suitable fungus growth is dissipated outside heat；Then, distributing with moisture, it is raw in mould, curved surface by
It gradually dries, forms confining bed, biological heat dissipation caused by more fungus strain fermentations is obstructed inside yeast, forms higher " top fire temperature
Degree ", fragrance component are also at this moment to start to generate.In order to reach the goals above, traditional handicraft, which dexterously takes, buckles, is micro-
Biological concentration, simultaneously room and closing heap etc. are tactful, and on the basis of being enriched with naturally, the space density by gradually increasing knee-piece improves bent
Temperature has reached the natural purifying of fungus strain and enzyme system, and the formation of bent flavor substance processed.Recent studies indicate that in white wine
Multiple types compound have healthy functions, be especially in the presence of nitrogenous compound-Tetramethylpyrazine class chemical combination in white wine
Object；In traditional yeast-making technology, such material demand synthesizes under the high temperature conditions (60 DEG C of ﹥).But in view of higher song Fang Wen
Degree, it is biggish turn over bent labor intensity, and complicated process adjustments cause product quality more difficult to control, cause four in high-temperature daqu
The synthetic effect of methylpyrazine is unsatisfactory.In recent years, it in yeast preparation process, is crushed to buckle from raw material and has realized machinery
Change, but subsequent technique remains in traditional-handwork operation level.So how to reduce starter-making temperature, reduce labor intensity
Under the premise of, the yeast of high Tetramethylpyrazine content is produced, the key issues of industry research are become.
Before this, inventor utilizes bacillus licheniformis, bacillus subtilis, Bacillus circulans, solution Bacillus stearothermophilus
Four kinds of bacterium take individually and hybrid bacterial strain training mode carry out producing Tetramethylpyrazine experiment, respectively although the results show that these bacterium
The yield of strain laboratory Tetramethylpyrazine is higher, but in high-temperature daqu training mode, Tetramethylpyrazine content is only medium temperature frame
1/10 to the 1/5 of formula yeast training mode is horizontal, far from meeting the needs of industrial production function yeast.Therefore, one plant is screened
Can be produced using medium temperature posture the bacillus licheniformis of high Tetramethylpyrazine content yeast very it is necessary to.
Summary of the invention
In order to reduce starter-making temperature and labor intensity, yeast Tetramethylpyrazine content is improved, the present invention is intended to provide a kind of
Produce bacillus licheniformis and its application of Tetramethylpyrazine.The present invention has high Tetramethylpyrazine synthesis capability for what screening obtained
Bacillus licheniformis after initial-stage culture, be applied to optimization after shelf daqu culture process in.Due in fermentation process
Mid-early stage keeps suitable starter-making temperature, so that functional microorganism is synthesized more Tetramethylpyrazine precursor, through 28d medium temperature (﹤ 50
DEG C) Qu-making process, Tetramethylpyrazine content reaches 400mg/kg.
The present invention produces the bacillus licheniformis of Tetramethylpyrazine, and classification naming is bacillus licheniformis GJBL055
(Bacillus licheniformis GJBL055), has been preserved in China typical culture collection center, the deposit date is
On April 24th, 2017, deposit number are CCTCC NO:M 2017204, and preservation address is Wuhan, China Wuhan University.
The present invention produces the application of the bacillus licheniformis of Tetramethylpyrazine, is the bacillus licheniformis that will produce Tetramethylpyrazine
It is applied in medium temperature shelf daqu culture process, produces the yeast of high Tetramethylpyrazine content.
The present invention produces the application of the bacillus licheniformis of Tetramethylpyrazine, includes the following steps:
Step 1: the preparation of function bacteria culture fluid
It is trained with the bacillus licheniformis GJBL055 bacterial strain that oese picking 1-2 environmental protection is stored in inclined-plane in 300-400mL liquid
It supports in base, 120rpm shaking table culture for 24 hours, obtains primary seed solution at 37 DEG C；Primary seed solution is inoculated with by 10% inoculum concentration
In fermentor, 80rpm culture for 24 hours, obtains secondary seed solution at 37 DEG C；Secondary seed solution is inoculated with by 10% inoculum concentration again
In fermentor, 80rpm culture for 24 hours, obtains three-level seed liquor, as function bacteria culture fluid at 37 DEG C；
10% inoculum concentration refers to that the volume of level-one or secondary seed solution accounts for the ratio of fermenter volume.
The fluid nutrient medium is constituted as follows by mass concentration:
Glucose 1wt%, yeast extract 1wt% (be purchased from Angel Yeast Co., Ltd), sodium chloride 0.5wt% are molten
Agent is water.
Step 2: the addition of function bacteria culture fluid
The function bacteria culture fluid that step 1 is obtained is added in the dedicated water storage tank of koji-making, and koji-making material whole wheat is added after mixing
Powder simultaneously stirs evenly, and is sufficiently mixed material by stepping on system and controls moisture content 36-37%, obtains curved billet, every piece of curved billet
Thickness control is 6.9-7.1cm (only needing to control thickness, the size of length and width does not have an impact for final result)；Function
The additive amount of bacteria culture fluid is 0.5% (i.e. the 0.5% of wholemeal gross mass) of every room into bent gross mass.
Step 3: the arrangement of functional shelf daqu
By the linear pendulum system of curved billet on bent frame, each song frame places five layers of curved billet (i.e. five layer frames, five layers of song), often
A song frame puts 32-34 block curved billet, each song frame spacing 5-8cm, and every room song frame puts into 3 rows 10 column.
Step 4: the culture of shelf daqu
Culture through the well laid shelf daqu of step 3 row includes the following six stage: sleeping bent receipts sweat phase, fermentation phase, low temperature
Cultivation phase, slow liter phase, high temperature flavouring phase and tempering humidity discharging phase, obtain the yeast of high Tetramethylpyrazine content.
4a, sleeping bent receipts sweat phase: equably spreading water in bent room before into song makes humidity >=80% in bent room, and humidity is conducive to
The enrichment of koji-making functional microorganism, control curved billet product temperature is identical as song room temperature holding after the song that crouched, curved billet surface after the song that crouched
It is tack-free, micro- whitening color.The sleeping bent receipts sweat phase closes the doors and windows, and controls bent Fang Wendu >=15 DEG C, the upper limit is environment temperature；Bent room humidity
>=80%.The sleeping bent time for receiving the sweat phase is 1.5-2.5 days.
4b, fermentation phase: entering the fermentation phase after the sleeping bent receipts sweat phase, the fermentation phase is 5-7 days, the liter for phase curved billet product temperature of fermenting
Warm speed control is at 4-6 DEG C/day；When the product temperature of upper layer curved billet reaches 43-46 DEG C, keep at such a temperature 12-20 hours it is laggard
Row turns over song for the first time.The song that turns over refers to that carry out lower layer exchanges with upper layer curved billet, middle part with external curved billet position.
Within the fermentation phase, when in curved billet surfaces of tacky and bent room it is micro- have sweet tea tart flavour when, Ying Jinhang humidity discharging is (so-called
Humidity discharging is the humidity reduced in bent room), humidity >=80% in bent room is controlled during humidity discharging, temperature declines≤5 DEG C, to prevent
Phenomena such as curved billet " dry to strip off the skin, crack " and " flu ".
4c, low temperature cultivation phase: the time of low temperature cultivation phase is 6-7 days, and the temperature of low temperature cultivation phase is controlled at 28-35 DEG C,
And daily temperature fluctuation range≤3 DEG C；At the 6th day of the low temperature cultivation phase or the 7th day, when curved billet product temperature reaches 35-38 DEG C,
Song is turned over for the second time.The bent end for terminating to represent the low temperature cultivation phase is turned over for the second time.
4d, slow liter phase: entering the slow liter phase after the low temperature cultivation phase, the heating rate for delaying liter phase curved billet product temperature is controlled in 3-
5 DEG C/day delay the phase of liter introversion room humid control in 60-80% until curved billet product temperature is warming up to 46-50 DEG C and is kept for 1-2 days.When
Curved billet maturity reaches 80-85%, and bent heart spring tide has been discharged, and feel is loose, and the bent heart reaches egg yolk size, and the slow liter phase terminates,
Start to pick up pile, into the high temperature flavouring phase.
4e, high temperature flavouring phase: it picks up the pile period of the day from 11 p.m. to 1 a.m and removes bent frame, curved billet is put into the type of being in line or isosceles triangle or linear type
It mixes and puts with isosceles triangle, to increase the tightness between curved billet, be conducive to the further raising of curved billet temperature and maintain；Pick up pile
The same day of son, 10, the control of curved billet product temperature in November at 45-50 DEG C, 3-9 month curved billet product temperature was controlled 47-52 DEG C of (other months
Temperature is lower, is not suitable for koji-making), and kept for 72 hours or more.
4f, tempering humidity discharging phase: after the high temperature flavouring phase, curved billet product temperature is reduced to room temperature from 50 DEG C or more naturally naturally,
Curved billet moisture content≤15% to get high Tetramethylpyrazine content yeast.
The beneficial effects of the present invention are embodied in:
1, voluntarily screening obtains Tetramethylpyrazine superior strain bacillus licheniformis to the present invention, which optimizes in laboratory
Under the conditions of produce Tetramethylpyrazine concentration reach 800mg/kg or more.
2, the present invention for the first time combines Tetramethylpyrazine Producing Strain with shelf daqu training mode, by inoculum concentration and
The optimization of shelf song posture model finds the training mode of shelf song, is more advantageous to the cell growth and four of bacillus licheniformis
The synthesis of methylpyrazine precursor.
3, the present invention constructs functional yeast posture medium temperature culture control model for the first time, the culture temperature in specific several stages
Degree, to realize that optimal yeast fungus strain, enzyme system, flavor substance system are laid a good foundation.By 28d medium temperature (50 DEG C of ﹤) koji-making, most
Tetramethylpyrazine content is increased to 400mg/kg or more in whole yeast.
The present invention combines modern biotechnology with traditional yeast production technology, by the lichens with independent intellectual property rights
Bacillus is applied in the posture daqu of middle temperature manufacturing process after optimization, produces and is rich in tetramethyl pyrrole with high temperature style of song lattice
The yeast of piperazine is also established to further increase white wine health active material for the mechanization, automation, intelligence of yeast production
There are a far-reaching social and economic implications on basis to the transformation and upgrade and Ecomanufacturing of realizing conventional industries.
Detailed description of the invention
Fig. 1 is the growing state of bacillus licheniformis GJBL055 bacterial strain at different temperatures.This hair as seen from Figure 1
Bright bacillus licheniformis GJBL055 bacterial strain remains to normal growth at 55 DEG C, and when temperature reaches 60 DEG C, thallus cannot normally give birth to
It is long, so fermentation phase temperature is not above 55 DEG C.
Specific embodiment
Embodiment 1: the preparation of bacillus licheniformis function bacteria culture fluid
It is trained with the bacillus licheniformis GJBL055 bacterial strain that oese picking 1-2 environmental protection is stored in inclined-plane in 300-400mL liquid
It supports in base, 120rpm shaking table culture for 24 hours, obtains primary seed solution at 37 DEG C；Primary seed solution is inoculated with by 10% inoculum concentration
In fermentor, 80rpm culture for 24 hours, obtains secondary seed solution at 37 DEG C；Secondary seed solution is inoculated with by 10% inoculum concentration again
In fermentor, 80rpm culture for 24 hours, obtains three-level seed liquor, as function bacteria culture fluid at 37 DEG C；
10% inoculum concentration refers to that the volume of level-one or secondary seed solution accounts for the ratio of fermenter volume.
The fluid nutrient medium is constituted as follows by mass concentration:
Glucose 1wt%, yeast extract 1wt% (be purchased from Angel Yeast Co., Ltd), sodium chloride 0.5wt% are molten
Agent is water.
Embodiment 2: the optimization of bacillus licheniformis function bacteria culture fluid additive amount
The function bacteria culture fluid that embodiment 1 obtains is added with the adding proportion of every room into Qu Zhiliang 0.1%, 0.5%, 1%
It adds in water storage tank, koji-making material wholemeal is added after mixing, mix, cultivated by the training method of shelf daqu.
When 1 Different adding amount of table, Tetramethylpyrazine yield
| Function bacterium solution additive amount | Fang Quzhong Tetramethylpyrazine content ppm out |
| 0.1% | 106.3 |
| 0.5% | 389.4 |
| 1% | 356.4 |
As shown in Table 1, when bacillus licheniformis function bacteria culture fluid adding proportion is 0.5%, Fang Quzhong tetramethyl pyrrole out
Piperazine content is higher.
Embodiment 3: the arrangement of functional shelf daqu
The function bacteria culture fluid that embodiment 1 obtains is added in water storage tank with the ratio of every room into Qu Zhiliang 0.5%, is mixed
Koji-making material wholemeal is added after even, be sufficiently mixed material by stepping on system and controls moisture content 36-37%, acquisition curved billet,
The thickness control of every piece of curved billet is in 6.9-7.1cm；It is placed in curved billet is linear on bent frame, each shelf puts 32-34 block
Curved billet, each 5-8 centimeters of frame spacing of song, is arranged three kinds altogether and puts scheme, is " five layer frames, six layers of song ", " five layer frames respectively
Five layers of song (bottom is not placed) ", " four layer frames, five layers of song ", every room puts 3 rows 10 altogether and arranges bent frame.
When 2 shelf daqu difference of table is arranged, room song Tetramethylpyrazine content out
| Three kinds are put scheme | Fang Quzhong Tetramethylpyrazine content ppm out |
| Five layer frames, six layers of song (common shelf daqu arrangement) of son | 276.3 |
| Five layer frames, five layers of song of son | 413.4 |
| Four layer frames, five layers of song of son | 306.4 |
As shown in Table 2, when the arrangement of shelf daqu is " five layer frames son five layers of song (bottom is not placed) ", Fang Quzhong tetramethyl out
Base levels of pyrazine is higher.
Embodiment 4: the optimization of the culture process of shelf daqu
The present invention is to the optimization of shelf daqu culture process mainly in the fermentation phase.Bacterial strain lichens gemma bar is used according to the present invention
The growth characteristics of bacterium at different temperatures, and fermentation phase temperature different in actual production go out the Tetramethylpyrazine of Fang Quzhong
Content, it is determined that optimal fermentation phase temperature.
Table 3 ferments phase different temperature control, out Fang Quzhong Tetramethylpyrazine content
| Fermentation phase temperature | Fang Quzhong Tetramethylpyrazine content ppm out |
| 40-45℃ | 246.3 |
| 45-50℃ | 423.4 |
| 50-60℃ | 0.32 |
For the control of fermentation phase temperature at 45-50 DEG C, Fang Quzhong Tetramethylpyrazine content is higher out as can be seen from Table 3.And
When temperature control is between 50-60 DEG C, since thallus is unable to normal growth, the synthesis of Tetramethylpyrazine can not be carried out.
At a temperature of 4 different fermentations of table, total plate count in knee-piece.
| Fermentation phase temperature | Total plate count (cfu/g) in knee-piece |
| 40-45℃ | 2.5*108 |
| 45-50℃ | 1.55*108 |
| 50-60℃ | 4.25*103 |
Embodiment 5: the culture of shelf daqu
Culture through the well laid shelf daqu of 3 row of embodiment (five layer frames, five layers of song) includes the following six stage: sleeping song
Sweat phase, fermentation phase, low temperature cultivation phase, slow liter phase, high temperature flavouring phase and tempering humidity discharging phase are received, high Tetramethylpyrazine content is obtained
Yeast.
4a, sleeping bent receive the sweat phase: equably spreading 50 kilograms of water purification before into song in bent room, make humidity in bent room >=
80%, in the case where opening wide door and window, the side curl that crouches is dried in the air tide, and humidity is conducive to the enrichment of koji-making functional microorganism.It has crouched and has controlled after song
Koji-making made-up article temperature is identical as bent room temperature holdings, but when bent room temperature is lower than 15 DEG C, the requirement of curved billet product temperature be maintained at 15 DEG C with
On.Curved billet surface after song of having crouched is tack-free, micro- whitening color.The sleeping bent receipts sweat phase closes the doors and windows, and controls bent Fang Wendu >=15 DEG C, the upper limit
For environment temperature；Bent room humidity >=80%.The sleeping bent receipts sweat phase can be by way of in curved billet all around careless grid, upper cap Lu Seats
Tide is protected in heat preservation, and when weather is higher compared with hot cell temperature, upper layer can not lid reed Seats.
4b, fermentation phase: entering the fermentation phase after the sleeping bent receipts sweat phase, the fermentation phase is 5-7 days, the heating rate control for phase of fermenting
It makes and (is stepped up in a few days ago the 1st, 2 day of phase (ferment) temperature of fermentation phase to 22~28 DEG C, third day temperature in 5 DEG C/day
Degree is stepped up to 35~38 DEG C), curved billet can be controlled by way of opening a window and opening reed Seats and delays Wen Shangsheng, to prevent heating
The too quickly or red Pi Xianxiang of appearance；Terminate a few days ago in the fermentation phase, Yao Liyong door and window strict control curved billet product temperature, when upper layer curved billet
Product temperature reach 43-46 DEG C, carry out turning over song for the first time after being kept for 12-20 hour at such a temperature, by lower layer and upper layer curved billet, in
Portion and the position of external curved billet swap.
Within the fermentation phase, when in curved billet surfaces of tacky and bent room it is micro- have sweet tea tart flavour when, Ying Jinhang humidity discharging, humidity discharging
Humidity >=80% in bent room is controlled in the process, and temperature declines≤5 DEG C, to prevent curved billet " dry strip off the skin, crack " and " flu " etc. from showing
As.
4c, low temperature cultivation phase: the time of low temperature cultivation phase is 6-7 days, and the temperature of low temperature cultivation phase is controlled at 28-35 DEG C,
And daily temperature fluctuation range≤3 DEG C；At the 6th day of the low temperature cultivation phase or the 7th day, when curved billet product temperature reaches 35-38 DEG C,
Song is turned over for the second time, and lower layer and upper layer curved billet, middle part and the position of external curved billet are swapped.
4d, slow liter phase: entering the slow liter phase after the low temperature cultivation phase, the heating rate for delaying liter phase curved billet product temperature is controlled in 3-
5 DEG C/day, until curved billet product temperature is warming up to 46-50 DEG C and is kept for 1-2 days, the slow phase introversion room humid control that rises, can in 60-80%
To carry out humidity discharging by door, window gap.During control temperature slow liter, first with door, window and to shake the head, recycle grass
Grid；When curved billet maturity reaches 80-85%, bent heart spring tide has been discharged, and feel is loose, and the bent heart reaches egg yolk size, delays the phase of liter
Terminate, starts to pick up pile, into the high temperature flavouring phase.
4e, high temperature flavouring phase: it picks up the pile period of the day from 11 p.m. to 1 a.m and removes bent frame, curved billet is put into the type of being in line or isosceles triangle or linear type
It mixes and puts with isosceles triangle, to increase the tightness between curved billet, be conducive to the further raising of curved billet temperature and maintain；It puts
Curved billet puts 6 layers altogether up and down when curved billet, and lower tight upper pine, gap is in Pagoda-shaped, and in favor of humidity discharging, surrounding encloses careless grid more and keeps high
Temperature；Picked up the same day of pile, 10, the control of curved billet product temperature in November at 45-50 DEG C, 3-9 month curved billet product temperature is controlled at 47-52 DEG C,
And kept for 72 hours or more.
4f, tempering humidity discharging phase: after the high temperature flavouring phase, curved billet product temperature drops to room temperature, curved billet from 50 DEG C or more naturally
Moisture content≤15% to get high Tetramethylpyrazine content yeast.
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not intended to limit the invention, any to be familiar with this skill
The people of art can do various change and modification, therefore protection model of the invention without departing from the spirit and scope of the present invention
Enclosing be subject to what claims were defined.
Claims (7)
1. a kind of bacillus licheniformis for producing Tetramethylpyrazine, it is characterised in that: the classification naming of the bacillus licheniformis is
Bacillus licheniformis GJBL055 (Bacillus licheniformis GJBL055) has been preserved in Chinese Typical Representative culture guarantor
Hiding center, the deposit date is on April 24th, 2017, deposit number was CCTCC NO:M 2017204, and preservation address is that China is military
Chinese Wuhan University.
2. a kind of application of the bacillus licheniformis described in claim 1 for producing Tetramethylpyrazine, it is characterised in that: be that will produce four
The bacillus licheniformis of methylpyrazine is applied in medium temperature shelf daqu culture process, produces the big of high Tetramethylpyrazine content
Song specifically comprises the following steps:
Step 1: the preparation of function bacteria culture fluid
The bacillus licheniformis GJBL055 bacterial strain on inclined-plane is stored in 300-400mL fluid nutrient medium with oese picking 1-2 environmental protection
In, 120rpm shaking table culture for 24 hours, obtains primary seed solution at 37 DEG C；Primary seed solution is inoculated in hair by 10% inoculum concentration
In fermentation tank, 80rpm culture for 24 hours, obtains secondary seed solution at 37 DEG C；Secondary seed solution is inoculated in hair by 10% inoculum concentration again
In fermentation tank, 80rpm culture for 24 hours, obtains three-level seed liquor, as function bacteria culture fluid at 37 DEG C；
Step 2: the addition of function bacteria culture fluid
The function bacteria culture fluid that step 1 is obtained is added in the dedicated water storage tank of koji-making, and koji-making material wholemeal is added simultaneously after mixing
It stirs evenly, be sufficiently mixed material by stepping on system and control moisture content 36-37%, obtain curved billet, the thickness of every piece of curved billet
Control is in 6.9-7.1cm；
Step 3: the arrangement of functional shelf daqu
By the linear pendulum system of curved billet on bent frame, each song frame places five layers of curved billet, each song frame spacing 5-8cm, Mei Gequ
Frame puts 32-34 block curved billet；
Step 4: the culture of shelf daqu
Culture through the well laid shelf daqu of step 3 row includes the following six stage: sleeping bent receipts sweat phase, fermentation phase, low temperature cultivation
Phase, slow liter phase, high temperature flavouring phase and tempering humidity discharging phase, obtain the yeast of high Tetramethylpyrazine content；Include the following steps:
4a, sleeping bent receipts sweat phase: equably spreading water in bent room before into song makes humidity >=80% in bent room, controls after the song that crouched bent
Made-up article temperature is identical as bent room temperature holding, and the bent receipts sweat phase of crouching controls bent Fang Wendu >=15 DEG C, and the upper limit is environment temperature, bent room humidity
>=80%；The sleeping bent time for receiving the sweat phase is 1.5-2.5 days；
4b, fermentation phase: entering the fermentation phase after the sleeping bent receipts sweat phase, the fermentation phase is 5-7 days, and the heating for phase curved billet product temperature of fermenting is fast
Degree control is at 4-6 DEG C/day；When the product temperature of upper layer curved billet reaches 43-46 DEG C, the is carried out after being kept for 12-20 hours at such a temperature
Once turn over song；
4c, low temperature cultivation phase: the time of low temperature cultivation phase is 6-7 days, and the temperature of low temperature cultivation phase is controlled at 28-35 DEG C, and
Daily temperature fluctuation range≤3 DEG C；At the 6th day of the low temperature cultivation phase or the 7th day, when curved billet product temperature reaches 35-38 DEG C, carry out
Song is turned over for the second time；
4d, slow rise the phase: enter after the low temperature cultivation phase it is slow rise the phase, delay rise the heating rate control of phase curved billet product temperature 3-5 DEG C/
It delays the phase of liter introversion room humid control in 60-80% until curved billet product temperature is warming up to 46-50 DEG C and is kept for 1-2 days；Work as curved billet
Maturity reaches 80-85%, and the slow liter phase terminates, and starts to pick up pile, into the high temperature flavouring phase；
4e, high temperature flavouring phase: it picks up the pile period of the day from 11 p.m. to 1 a.m and removes bent frame, curved billet is put into the type of being in line or isosceles triangle or linear type and product
Font mixing is put；Picked up the same day of pile, 10, the control of curved billet product temperature in November at 45-50 DEG C, the control of 3-9 month curved billet product temperature
At 47-52 DEG C, and kept for 72 hours or more；
4f, tempering humidity discharging phase: after the high temperature flavouring phase, curved billet product temperature is reduced to room temperature, curved billet from 50 DEG C or more naturally naturally
Moisture content≤15% to get high Tetramethylpyrazine content yeast.
3. application according to claim 2, it is characterised in that:
In step 1, the fluid nutrient medium is constituted as follows by mass concentration: glucose 1wt%, yeast extract 1wt%, sodium chloride
0.5wt%, solvent are water.
4. application according to claim 2, it is characterised in that:
In step 2, the additive amount of function bacteria culture fluid is every room into the 0.5% of bent gross mass.
5. application according to claim 2, it is characterised in that:
Fermentation phase and the interim song that turns over of low temperature cultivation refer to the friendship for carrying out lower layer and upper layer curved billet, middle part and external curved billet position
It changes.
6. application according to claim 2, it is characterised in that:
Within the fermentation phase, when in curved billet surfaces of tacky and bent room it is micro- have sweet tea tart flavour when, Ying Jinhang humidity discharging, humidity discharging process
Humidity >=80% in the bent room of middle control, temperature decline≤5 DEG C.
7. application according to claim 2, it is characterised in that:
The temperature of fermentation phase controls≤55 DEG C.
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| CN104962432A (en) * | 2015-07-29 | 2015-10-07 | 安徽古井贡酒股份有限公司 | Room culture method of medium-high temperature yeast |
| CN105002115A (en) * | 2015-07-28 | 2015-10-28 | 贵州茅台酒股份有限公司 | Bacillus licheniformis, yeast prepared through same and preparation method of yeast |
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| CN105002115A (en) * | 2015-07-28 | 2015-10-28 | 贵州茅台酒股份有限公司 | Bacillus licheniformis, yeast prepared through same and preparation method of yeast |
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