CN107446860A - A kind of bacillus licheniformis for producing Tetramethylpyrazine and its application - Google Patents

A kind of bacillus licheniformis for producing Tetramethylpyrazine and its application Download PDF

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CN107446860A
CN107446860A CN201710788378.6A CN201710788378A CN107446860A CN 107446860 A CN107446860 A CN 107446860A CN 201710788378 A CN201710788378 A CN 201710788378A CN 107446860 A CN107446860 A CN 107446860A
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temperature
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curved billet
daqu
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CN107446860B (en
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葛向阳
何宏魁
李晓欢
张严
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Anhui Ruisiweier Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12G3/02—Preparation of other alcoholic beverages by fermentation

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Abstract

The invention discloses a kind of bacillus licheniformis for producing Tetramethylpyrazine and its application, the Classification And Nomenclature of wherein described bacillus licheniformis is bacillus licheniformis GJBL055 (Bacillus licheniformis GJBL055), it has been preserved in China typical culture collection center, preservation date is on April 24th, 2017, and deposit number is CCTCC NO：M 2017204, preservation address are Wuhan, China Wuhan University.Use medium temperature shelf training mode, the bacillus licheniformis of the present invention is linked into flat board Daqu, pass through the preculture to functional microorganism, the structure of shelf song spatial model, the four stages regulation and control of area room temperature, a kind of Daqu manufacturing process of high yield Tetramethylpyrazine under medium temperature condition is formed, Tetramethylpyrazine content in Daqu is brought up to 400mg/kg.

Description

A kind of bacillus licheniformis for producing Tetramethylpyrazine and its application
Technical field
The present invention relates to a kind of bacillus licheniformis for producing Tetramethylpyrazine and its application, belongs to fermentation engineering neck Domain.
Background technology
" song is the bone of wine ", Daqu plays vital effect in liquor production.It prepares purpose：Multi-functional bacterium The formation of system, multi-enzyme system and a variety of bouquet principles matter.In Daqu production technology, bacterium and saccharomycete ferment first, make song Heart temperature is raised, and the top layer for forming suitable fungus growth is dissipated outside heat；Then, distributing with moisture, it is raw in mould, curved surface by Gradually dry, form confining bed, biological heat produced caused by more fungus strain fermentations is obstructed inside Daqu, forms higher " top fire temperature Degree ", fragrance component be also at this moment start caused by.In order to reach object above, traditional handicraft, which dexterously takes, buckles, is micro- Biological concentration and the strategy such as room and closing heap, on the basis of being enriched with naturally, the space density by gradually increasing knee-piece improves bent Temperature, the natural purifying of fungus strain and enzyme system, and the formation of bent flavor substance processed are reached.Recent studies indicate that in white wine Multiple types compound there are healthy functions, be especially in the presence of nitrogen-containing compound-Tetramethylpyrazine class chemical combination in white wine Thing；In traditional yeast-making technology, (60 DEG C of ﹥) is synthesized such material demand under the high temperature conditions.But in view of higher bent room temperature Degree, it is larger turn over bent labor intensity, and complicated process adjustments cause the more difficult control of product quality, cause four in high-temperature daqu The synthetic effect of methylpyrazine is unsatisfactory.In recent years, in Daqu preparation technology, it 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 On the premise of, the Daqu of high Tetramethylpyrazine content is produced, turns into the emphasis problem of industry research.
Before this, inventor utilizes bacillus licheniformis, bacillus subtilis, Bacillus circulans, solution Bacillus stearothermophilus Four kinds of bacterium, take respectively individually and hybrid bacterial strain training mode carries out producing Tetramethylpyrazine experiment, as a result show, although 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 Daqu training mode is horizontal, far from meeting the needs of industrial production function Daqu.Therefore, one plant is screened Medium temperature posture can be used, which to produce the bacillus licheniformis of high Tetramethylpyrazine content Daqu, to be extremely necessary.
The content of the invention
In order to reduce starter-making temperature and labor intensity, Daqu 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 by 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, functional microorganism is synthesized more Tetramethylpyrazine precursor, through 28d medium temperatures (﹤ 50 DEG C) Qu-making process, Tetramethylpyrazine content reaches 400mg/kg.
The bacillus licheniformis of present invention production Tetramethylpyrazine, Classification And Nomenclature is bacillus licheniformis GJBL055 (Bacillus licheniformis GJBL055), has been preserved in China typical culture collection center, preservation date is On April 24th, 2017, deposit number are CCTCC NO：M 2017204, preservation address are Wuhan, China Wuhan University.
The application of the bacillus licheniformis of present invention production Tetramethylpyrazine, it is by the bacillus licheniformis of production Tetramethylpyrazine It is applied in medium temperature shelf daqu culture process, produces the Daqu of high Tetramethylpyrazine content.
The application of the bacillus licheniformis of present invention production Tetramethylpyrazine, comprises the following steps：
Step 1：The preparation of function bacteria culture fluid
The bacillus licheniformis GJBL055 bacterial strains that inclined-plane is stored in oese picking 1-2 environmental protection are trained in 300-400mL liquid Support in base, 120rpm shaking table cultures 24h at 37 DEG C, obtain primary seed solution；Primary seed solution is inoculated with by 10% inoculum concentration In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains secondary seed solution；Secondary seed solution is inoculated with by 10% inoculum concentration again In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains three-level seed liquor, as function bacteria culture fluid；
10% inoculum concentration refers to that the volume of one-level or secondary seed solution accounts for the ratio of fermenter volume.
The fluid nutrient medium is formed as follows by mass concentration：
Glucose 1wt%, yeast extract 1wt% (are purchased from Angel Yeast Co., Ltd), sodium chloride 0.5wt%, 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 special water storage tank of koji-making, and koji-making material whole wheat is added after mixing Powder simultaneously stirs, and is 36-37% by stepping on system to be sufficiently mixed material and control moisture content, 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 addition of bacteria culture fluid is 0.5% (i.e. the 0.5% of wholemeal gross mass) for entering bent gross mass per room.
Step 3：The arrangement of feature shelf daqu
By the linear pendulum system of curved billet on bent frame, each bent frame places five layers of curved billet (i.e. five layer frames, five layers of song), often Individual bent frame puts 32-34 block curved billets, each bent frame spacing 5-8cm, and putting into 3 rows 10 per room song frame arranges.
Step 4：The culture of shelf daqu
The culture for the shelf daqu arranged through step 3 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 Daqu of high Tetramethylpyrazine content.
4a, sleeping bent receipts sweat phase：Equably spreading water in bent room before entering song makes humidity >=80% in bent room, and humidity is advantageous to The enrichment of koji-making functional microorganism, control after the song that crouched curved billet product temperature identical with bent room temperature holding, 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 room temperature >=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：The sleeping bent receipts sweat phase enters the fermentation phase after terminating, and 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 strata curved billet reaches 43-46 DEG C, holding 12-20 hours are laggard at such a temperature Row turns over song for the first time.The song that turns over refers to that carry out lower floor exchanges with upper strata curved billet, middle part with outside curved billet position.
It is micro- when have sweet tea tart flavour when curved billet surfaces of tacky, and in bent room within the fermentation phase, it is (so-called humidity discharging should to be carried out Humidity discharging is to reduce the humidity 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, ftracture " and " flu ".
4c, low temperature cultivation phase：The time of low temperature cultivation phase be 6-7 days, the temperature control of low temperature cultivation phase 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：The low temperature cultivation phase enters the slow liter phase after terminating, and 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 kept for 1-2 days, delay and rise phase introversion room humid control in 60-80%.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：Pick up the pile period of the day from 11 p.m. to 1 a.m and remove bent frame, curved billet is put into the type of being in line either isosceles triangle or linear pattern Mix and put with isosceles triangle, to increase the tight ness rating between curved billet, be advantageous to the further rise of curved billet temperature with maintaining；Pick up pile The same day of son, 10, curved billet product temperature control in November at 45-50 DEG C, 3-9 month curved billets product temperature was controlled 47-52 DEG C of (other months Temperature is relatively low, is not suitable for koji-making), and kept for more than 72 hours.
4f, tempering humidity discharging phase：After the high temperature flavouring phase terminates, curved billet product temperature is reduced to room temperature naturally from more than 50 DEG C naturally, Curved billet moisture content≤15%, produce the Daqu of high Tetramethylpyrazine content.
Beneficial effects of the present invention are embodied in：
1st, voluntarily screening obtains Tetramethylpyrazine superior strain bacillus licheniformis to the present invention, and the bacterial strain optimizes in laboratory Under the conditions of production Tetramethylpyrazine concentration reach more than 800mg/kg.
2nd, Tetramethylpyrazine Producing Strain is combined by the present invention with shelf daqu training mode first, by inoculum concentration and The optimization of shelf song posture model, the bent training mode of shelf is found, be more beneficial for the cell growth of bacillus licheniformis, and four The synthesis of methylpyrazine precursor.
3rd, the present invention constructs feature Daqu posture medium temperature culture control model first, the culture temperature in clear and definite several stages Degree, to realize that optimal Daqu fungus strain, enzyme system, flavor substance system are laid a good foundation.By 28d medium temperatures (50 DEG C of ﹤) koji-making, most Tetramethylpyrazine content brings up to more than 400mg/kg in whole Daqu.
Modern biotechnology is combined by the present invention with traditional Daqu production technology, by the lichens with independent intellectual property rights Bacillus is applied in the posture daqu of middle temperature manufacturing process after optimization, and production is rich in tetramethyl pyrrole with high temperature style of song lattice The Daqu of piperazine, further to improve white wine health active material, also established for the mechanization, automation, intellectuality of Daqu production The transformation and upgrade and Ecomanufacturing for realizing conventional industries are had far-reaching society and economic implications by basis.
Brief description of the drawings
Fig. 1 is the growing state of bacillus licheniformis GJBL055 bacterial strains at different temperatures.This hair as seen from Figure 1 Bright bacillus licheniformis GJBL055 bacterial strains remain to normal growth at 55 DEG C, and when temperature reaches 60 DEG C, thalline can not normally give birth to It is long, so fermentation phase temperature is not above 55 DEG C.
Embodiment
Embodiment 1：The preparation of bacillus licheniformis function bacteria culture fluid
The bacillus licheniformis GJBL055 bacterial strains that inclined-plane is stored in oese picking 1-2 environmental protection are trained in 300-400mL liquid Support in base, 120rpm shaking table cultures 24h at 37 DEG C, obtain primary seed solution；Primary seed solution is inoculated with by 10% inoculum concentration In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains secondary seed solution；Secondary seed solution is inoculated with by 10% inoculum concentration again In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains three-level seed liquor, as function bacteria culture fluid；
10% inoculum concentration refers to that the volume of one-level or secondary seed solution accounts for the ratio of fermenter volume.
The fluid nutrient medium is formed as follows by mass concentration：
Glucose 1wt%, yeast extract 1wt% (are purchased from Angel Yeast Co., Ltd), sodium chloride 0.5wt%, molten Agent is water.
Embodiment 2：The optimization of bacillus licheniformis function bacteria culture fluid addition
The adding proportion that function bacteria culture fluid that embodiment 1 obtains is entered to Qu Zhiliang 0.1%, 0.5%, 1% with every room adds Add in water storage tank, koji-making material wholemeal is added after mixing, mix, cultivated by the training method of shelf daqu.
During 1 Different adding amount of table, Tetramethylpyrazine yield
Function bacterium solution addition Go out Tetramethylpyrazine content ppm in room song
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%, tetramethyl pyrrole in room song is gone out Piperazine content is higher.
Embodiment 3：The arrangement of feature shelf daqu
The ratio that function bacteria culture fluid that embodiment 1 obtains is entered to Qu Zhiliang 0.5% with every room is added in water storage tank, is mixed Koji-making material wholemeal is added after even, is 36-37% by stepping on system to be sufficiently mixed material and control moisture content, obtains 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 blocks Curved billet, each bent frame spacing 5-8 centimetres, set three kinds altogether and put scheme, be " 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 ", put 3 rows 10 per room and arrange bent frame altogether.
During the difference arrangement of the shelf daqu of table 2, go out room song Tetramethylpyrazine content
Three kinds are put scheme Go out Tetramethylpyrazine content ppm in room song
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, five layers of song (bottom is not placed) ", tetramethyl in room song is gone out Base levels of pyrazine is higher.
Embodiment 4：The optimization of the culture process of shelf daqu
Optimization of the present invention to shelf daqu culture process is 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 the Tetramethylpyrazine that different fermentation phase temperature in actual production goes out in room song Content, it is determined that optimal fermentation phase temperature.
Table 3 ferments phase different temperature control, goes out Tetramethylpyrazine content in room song
Fermentation phase temperature Go out Tetramethylpyrazine content ppm in room song
40-45℃ 246.3
45-50℃ 423.4
50-60℃ 0.32
It is higher to go out Tetramethylpyrazine content in room song at 45-50 DEG C for phase temperature control of fermenting as can be seen from Table 3.And Temperature control is when between 50-60 DEG C, because thalline is unable to normal growth, can not carry out the synthesis of Tetramethylpyrazine.
At a temperature of the different fermentations of table 4, 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
The culture for the shelf daqu (five layer frames, five layers of song) arranged through embodiment 3 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, obtains high Tetramethylpyrazine content Daqu.
4a, sleeping bent receipts sweat phase：Enter before song equably to spread 50 kilograms of water purification 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 advantageous to the enrichment of koji-making functional microorganism.Crouch and controlled after song Koji-making made-up article temperature is identical with bent room temperature holding, but when bent room temperature be less than 15 DEG C when, curved billet product temperature requirement 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 room temperature >=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 insulation, and when weather is higher compared with hot cell temperature, upper strata can not lid reed Seats.
4b, fermentation phase：The sleeping bent receipts sweat phase enters the fermentation phase after terminating, and the fermentation phase is 5-7 days, the programming rate control for phase of fermenting Make and (stepped up in 5 DEG C/day in a few days ago (the 1st, 2 day of the phase of fermenting) temperature of fermentation phase to 22~28 DEG C, the 3rd day temperature Degree is stepped up to 35~38 DEG C), the slow temperature of curved billet can be controlled to rise by way of opening a window and opening Lu Seats, to prevent heating The too quickly or red skin phenomenon of appearance；Terminate a few days ago in the fermentation phase, curved billet product temperature is strictly controlled using door and window, when upper strata curved billet Product temperature reach 43-46 DEG C, carry out turning over song for the first time after keeping 12-20 hours at such a temperature, by lower floor and upper strata curved billet, in Portion and the position of outside curved billet swap.
It is micro- when have sweet tea tart flavour when curved billet surfaces of tacky, and in bent room within the fermentation phase, humidity discharging, humidity discharging should be carried out During control humidity >=80% in bent room, temperature declines≤5 DEG C, to prevent curved billet " dry strip off the skin, ftracture " and " flu " etc. from showing As.
4c, low temperature cultivation phase：The time of low temperature cultivation phase be 6-7 days, the temperature control of low temperature cultivation phase 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 floor and upper strata curved billet, middle part and the position of outside curved billet are swapped.
4d, slow liter phase：The low temperature cultivation phase enters the slow liter phase after terminating, and 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 kept for 1-2 days, delay and rise phase introversion room humid control in 60-80%, can To carry out humidity discharging by door, window gap.During the slow liter of control temperature, 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, start to pick up pile, into the high temperature flavouring phase.
4e, high temperature flavouring phase：Pick up the pile period of the day from 11 p.m. to 1 a.m and remove bent frame, curved billet is put into the type of being in line either isosceles triangle or linear pattern Mix and put with isosceles triangle, to increase the tight ness rating between curved billet, be advantageous to the further rise of curved billet temperature with maintaining；Put Curved billet puts 6 layers altogether up and down during curved billet, and lower tight upper pine, space 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, curved billet product temperature control in November at 45-50 DEG C, 3-9 month curved billets product temperature is controlled at 47-52 DEG C, And kept for more than 72 hours.
4f, tempering humidity discharging phase：After the high temperature flavouring phase terminates, curved billet product temperature drops to room temperature, curved billet naturally from more than 50 DEG C Moisture content≤15%, produce the Daqu of high Tetramethylpyrazine content.
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this skill The people of art, without departing from the spirit and scope of the present invention, it can all do various change and modification, therefore the protection model of the present invention Enclose being defined of being defined by claims.

Claims (10)

  1. A kind of 1. bacillus licheniformis for producing Tetramethylpyrazine, it is characterised in that：The Classification And Nomenclature of the bacillus licheniformis is Bacillus licheniformis GJBL055 (Bacillus licheniformis GJBL055), Chinese Typical Representative culture guarantor has been preserved in it Tibetan center, preservation date are on April 24th, 2017, and deposit number is CCTCC NO：M 2017204, preservation address are military for China Chinese Wuhan University.
  2. A kind of 2. application of the bacillus licheniformis of the production Tetramethylpyrazine described in claim 1, it is characterised in that：It is 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 It is bent.
  3. 3. application according to claim 2, it is characterised in that comprise the following steps：
    Step 1：The preparation of function bacteria culture fluid
    The bacillus licheniformis GJBL055 bacterial strains on inclined-plane are stored in 300-400mL fluid nutrient mediums with oese picking 1-2 environmental protection In, 120rpm shaking table cultures 24h at 37 DEG C, obtain primary seed solution；Primary seed solution is inoculated in hair by 10% inoculum concentration In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains secondary seed solution；Secondary seed solution is inoculated in hair by 10% inoculum concentration again In fermentation tank, 80rpm cultivates 24h at 37 DEG C, obtains three-level seed liquor, as function bacteria culture fluid；
    Step 2：The addition of function bacteria culture fluid
    The function bacteria culture fluid that step 1 is obtained is added in the special water storage tank of koji-making, and koji-making material wholemeal is added simultaneously after mixing Stir, be 36-37% by stepping on system to be sufficiently mixed material and control moisture content, obtain curved billet, the thickness of every piece of curved billet Control is in 6.9-7.1cm；
    Step 3：The arrangement of feature shelf daqu
    By the linear pendulum system of curved billet on bent frame, each bent frame puts 32-34 block curved billets；
    Step 4：The culture of shelf daqu
    The culture for the shelf daqu arranged through step 3 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 Daqu of high Tetramethylpyrazine content.
  4. 4. application according to claim 3, it is characterised in that：
    In step 1, the fluid nutrient medium is formed as follows by mass concentration：Glucose 1wt%, yeast extract 1wt%, sodium chloride 0.5wt%, solvent are water.
  5. 5. application according to claim 3, it is characterised in that：
    In step 2, the addition of function bacteria culture fluid is to enter the 0.5% of bent gross mass per room.
  6. 6. application according to claim 3, it is characterised in that：
    In step 3, each bent frame places five layers of curved billet, each bent frame spacing 5-8cm.
  7. 7. application according to claim 3, it is characterised in that the incubation of shelf daqu comprises the following steps：
    4a, sleeping bent receipts sweat phase：Equably spreading water in bent room before entering song makes humidity >=80% in bent room, is controlled after the song that crouched bent Made-up article temperature is identical with bent room temperature holding, and the bent receipts sweat phase of crouching controls bent room temperature >=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：The sleeping bent receipts sweat phase enters the fermentation phase after terminating, and the fermentation phase is 5-7 days, the heating speed for phase curved billet product temperature of fermenting Degree control is at 4-6 DEG C/day；When the product temperature of upper strata curved billet reaches 43-46 DEG C, keep carrying out after 12-20 hours the at such a temperature Once turn over song；
    4c, low temperature cultivation phase：The time of low temperature cultivation phase be 6-7 days, the temperature control of low temperature cultivation phase 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 liter phase：The low temperature cultivation phase terminate after enter it is slow rise the phase, delay rise the heating rate control of phase curved billet product temperature 3-5 DEG C/ My god, until curved billet product temperature is warming up to 46-50 DEG C and kept for 1-2 days, delay and rise phase introversion room humid control in 60-80%；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：Pick up the pile period of the day from 11 p.m. to 1 a.m and remove bent frame, curved billet is put into the type of being in line either isosceles triangle or linear pattern and product Font mixing is put；Picked up the same day of pile, 10, curved billet product temperature control in November at 45-50 DEG C, the control of 3-9 month curved billets product temperature At 47-52 DEG C, and kept for more than 72 hours；
    4f, tempering humidity discharging phase：After the high temperature flavouring phase terminates, curved billet product temperature is reduced to room temperature, curved billet naturally from more than 50 DEG C naturally Moisture content≤15%, produce the Daqu of high Tetramethylpyrazine content.
  8. 8. application according to claim 7, it is characterised in that：
    Fermentation phase and the interim song that turns over of low temperature cultivation refer to carry out lower floor and upper strata curved billet, the friendship at middle part and outside curved billet position Change.
  9. 9. application according to claim 7, it is characterised in that：
    It is micro- when have sweet tea tart flavour when curved billet surfaces of tacky, and in bent room within the fermentation phase, humidity discharging, humidity discharging process should be carried out Humidity >=80% in the middle bent room of control, temperature decline≤5 DEG C.
  10. 10. application according to claim 7, it is characterised in that：
    Temperature control≤55 DEG C of fermentation phase.
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