CN103952316A - Fermentation Monascus strain and production process thereof - Google Patents
Fermentation Monascus strain and production process thereof Download PDFInfo
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Abstract
The invention discloses a fermentation Monascus strain, which has been preserved in China Center for type culture collection, Luojia mountain, Wuhan University, Wuchang, Wuhan, Hubei Province in February 23, 2011, and has an accession number of CCTCC NO:M 2011047. T The strain is prepared through steps of preservation of vacuum freezing tubes, preparation of test tube slant strains, preparation of triangular flask strains, preparation of seed tank seeds, seed dressing, heaping, spreading, fermentation culture and drying. Fermentation Monascus strain is a composite strain produced by biological fermentation, and rich enzyme systems can be produced by starter propagation fermentation. The process makes up for the defect of incomplete enzyme systems of single strain starter propagation, and can significantly improve vitality of enzyme systems protease, saccharifying enzyme and esterification enzyme of finished starter, and enhance protein utilization rate of materials and yield. The fermentation Monascus strain has strong color production capacity, produce shaematochrome and flavochrome in the fermentation process, so as to improve red index and yellow index of the product and endow the product with red and bright luster.
Description
Technical field
The present invention relates to a kind of technical field of bioengineering, particularly a kind of red colouring agent for food, also used as a Chinese medicine bacterial classification and production technique thereof of high-efficiency fermenting saccharogenic power.
Background technology
Saccharification flavoring yeast is domestic initiation specialty for making soy sauce, the compound sort of quyi of series of jam product and vinegar.It is the not congruent problem of single culture koji, enzyme activity for traditional zymotic jam product, the composite bacteria that utilizes modern biological engineering, screening, purifying, seed selection to improve, there is the stronger saccharogenic power of generation, esterification power, flavouring, whitening power and aspartic protease, mouthfeel, color and luster, the fragrance of soy sauce, vinegar and jam product be can obviously improve, and full nitrogen and the amino nitrogen content of finished product significantly improved.Fermentation red colouring agent for food, also used as a Chinese medicine is the chief component bacterial classification of saccharification flavoring yeast.
Summary of the invention
A kind of red colouring agent for food, also used as a Chinese medicine bacterial classification and production technique thereof of high-efficiency fermenting saccharogenic power the object of the invention is in order to be provided.
A fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification, this bacterial classification is preserved in Wuhan, Hubei loujia hill belongs Wuhan University Chinese Typical Representative culture collection center on February 23rd, 2011, and preserving number is: CCTCC NO:M2011047.Classification And Nomenclature: Monascus fulginosus JCFM-1, Monascus fuliginosus JCFM-1.
The ferment production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification, comprises the following steps:
Step 1, vacuum freezing pipe at 4 ℃, the preservation of normal temperature refrigerator;
Step 2, the preparation of test tube slant bacterial classification:
One, conditioned media: peptone 2%, Zulkovsky starch 2%, Fructus Hordei Germinatus soak powder 2%, (NH
4)
2sO
40.1%, MgSO
40.1%, K
2hPO
40.1%, agar 2%, natural pH; With 18 * 180 (mm) test tube, every pipe dress substratum 7～10ml (substratum is hot melt fully), with tampon, seal, 10 test tube bag a bundles, vertically be put in iron wire frame, 0.11MPa sterilizing 30min, is cooled to 50～60 ℃, incline inclined-plane processed (require slant medium length be test tube 3/4), after condensation, wrap up bundled; Cultivate 48hr, on inspection without miscellaneous bacteria, can use for 30 ℃;
Two, inoculation culture:
1, vacuum freezing ampoul tube turns test tube slant bacterial classification: with aseptic capillary pipet, one pipe stroke-physiological saline solution is added in ampoul tube, stir evenly, bacterium liquid is sucked to capillary pipet, in access slant tube; Cultivate 3～5 days for 30 ℃, switching activated for 2 to 3 generations, cultivated 8～10 days, put 4 ℃, refrigerator and save backup for 30 ℃;
2, slant tube strain transfer: get a slant tube kind, wipe examination totally with 75% spirituous solution, access 20～30 test tube slants with aseptic bamboo let in super clean bench, on spirit lamp flame; It is identical that cultivation and store method and described vacuum freezing ampoul tube turn test tube slant bacterial classification;
Step 3, the preparation of liquid triangle shaking flask bacterial classification:
One, substratum: glucose 4%, NaNO
30.2%, MgSO
40.1%, extractum carnis 0.3%, peptone 0.8%, yeast extract paste 0.5%, pH5.0; The 500ml capacity triangular flask loading amount of take is 100ml, and ground rice is weighed respectively to dress, and inorganic salt unified preparation dissolve, then are sub-packed in each triangular flask, fill in tampon, with kraft paper, wrap up.0.11MPa, sterilizing 30min, cooling standby;
Two, inoculation, cultivation: get a test tube slant bacterial classification, with aseptic bamboo let, access in 3～5 liquid triangle shake-flask culture bases, in 30 ℃, the about 48h of 200rpm shaking table shaking culture, hand over workshop to check and accept standby;
Step 4, the preparation of seeding tank liquid seeds:
1, nutrient solution formula: rice meal 7.5%, NaNO
30.4%, MgSO
40.1%, K
2hPO
40.1%, soya-bean oil 0.1%; With 75% alcohol, clean visor mouth to be opened, in the situation that guaranteeing seeding tank malleation, unclamp seeding tank visor, with Alcohol Flame circle, seal visor mouth taking off visor simultaneously, then in flame ring, open triangle bottleneck, pour seed in triangular flask into seeding tank rapidly, remove flame ring simultaneously, cover visor, obturage;
2, culture condition and control: rotating speed: 240rpm, temperature: 30～32 ℃, tank pressure: 0.05～0.07MPa, air flow quantity: 8～10m
3/ hr, incubation time: approximately 48 hours;
Step 5, fermentation culture:
Rice dipping, drip washing, drain, steam rice: the rice draining is steamed to 2～3min at the temperature of 102 ℃;
The preparation of bacterial classification: by rice amount 16% (15%5-2+1%A
1) inoculum size takes bacterial classification, the Glacial acetic acid of amount of rice 1% before inoculation, mixes rear standby;
Load weighted bacterial classification is evenly sprinkled upon on the rice of spreading for cooling, turns 3 times by hand, guarantee that bacterial classification fully mixes with material.The uniform bank of inoculation is overlayed to one end of woollen blanket under gauze, pile the thick 40～50cm of being, woollen blanket is added a cover on surface.
Step 6, Da Dui: be divided into that high hot house is beaten heap and Quchi is beaten heap, room temperature during lower than 20 ℃ high hot house beat heap; Room temperature during higher than 20 ℃ Quchi beat heap, pile thick 35～40cm, surface, coated with the felt insulation of sterilizing, keeps 20～30 ℃ of room temperatures, 15～20hr;
Step 7, stand heap are grown flower: comprise that high hot house material stand heap is grown flower and Quchi material stand heap is grown flower, 45～48 ℃ of stand heaps, keep 20～30 ℃ of room temperatures, and product temperature maintains 32～35 ℃, 20hr;
Step 8, Jia Shui: keep material moisture approximately 35%～55%, room temperature, at 20～25 ℃, is cultivated 4 days;
Step 9, go out pond and dry.
Beneficial effect of the present invention: the application of esterified red yeast in white wine is produced can effectively improve the esterification power of Daqu, promote that organic acid changes into corresponding ester, and it is to caproic acid and acetic acid changes into ethyl hexanoate and ethyl acetate has very strong selectivity, simultaneously, because of its acid resistance stronger, make organic acid change into corresponding ester, removed the toxic action of acid to cell; The proteolytic enzyme of its generation also can rich liquor flavour ingredient.Ester conversion rate is more than 95%, and esterification power >=35mg/g can accelerate the esterification of organic acid and alcohol in white wine production application, promote to generate various ester classes, shorten liquor fermentation period, increase ethyl hexanoate etc. is the synthesis capability of fragrant class material, significantly improves the quality percentage of white wine.
Accompanying drawing explanation
In order to be illustrated more clearly in technical scheme of the present invention, below the accompanying drawing of required use during the present invention is described is briefly described.
Fig. 1 is implementing process schema of the present invention;
Embodiment
A kind of fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification described in the present embodiment, this bacterial classification is preserved in Wuhan, Hubei loujia hill belongs Wuhan University Chinese Typical Representative culture collection center on February 23rd, 2011, and preserving number is: CCTCC NO:M2011047.
As shown in Figure 1, a kind of production technique of the red colouring agent for food, also used as a Chinese medicine bacterial classification that ferments, comprises the following steps:
S1, vacuum freezing pipe.4 ℃, the preservation of normal temperature refrigerator.
S2, the preparation of test tube slant bacterial classification.
One, substratum: peptone 2%, Zulkovsky starch 2%, Fructus Hordei Germinatus soak powder 2%, (NH
4)
2sO
40.1%, MgSO
40.1%, K
2hPO
40.1%, agar 2%, natural pH.
Method: with 18 * 180 (mm) test tube, every pipe dress substratum 7～10ml (substratum is hot melt fully), with tampon, seal, 10 test tube bag a bundles, vertically be put in iron wire frame, 0.11MPa sterilizing 30min, is cooled to 50～60 ℃, incline inclined-plane processed (require slant medium length be test tube 3/4), after condensation, wrap up bundled; Cultivate 48hr, on inspection without miscellaneous bacteria, can use for 30 ℃.
Two, inoculation culture:
1, vacuum freezing ampoul tube turns test tube slant bacterial classification.
Ampoul tube is led out, in super clean bench, with 75% cotton ball soaked in alcohol, wiped examination totally, at the inclined to one side top of ampoul tube, with the standardized circle wheel of emery wheel trace, with sterilizing newspaper or kraft paper, wrap up its two ends, on spirit lamp flame, be broken.More difficult if fracture, can take following measures:
A, in spirit lamp flame top, with aseptic nipper, its top is broken into pieces;
B, with spirit lamp flame calcination ampoul tube top, with aseptic capillary pipet, drip sterilized water in calcination place, it is burst.
Then with aseptic capillary pipet, one pipe stroke-physiological saline solution is added in ampoul tube, stir evenly, bacterium liquid is sucked to capillary pipet, in access slant tube.A general ampoul tube bacterial classification can connect 3～5 of test tube slants.Cultivate 3～5 days for 30 ℃, switching activated for 2 to 3 generations, cultivated 8～10 days, put 4 ℃, refrigerator and save backup for 30 ℃.
2, slant tube strain transfer
Get a slant tube kind, with 75% spirituous solution, wipe examination totally, in super clean bench, on spirit lamp flame, with aseptic bamboo let, access 20～30 test tube slants.Cultivate with store method and turn test tube slant bacterial classification with vacuum freezing ampoul tube.
S3, the preparation of liquid triangle shaking flask bacterial classification.
One, substratum: glucose 4%, NaNO
30.2%, MgSO
40.1%, extractum carnis 0.3%, peptone 0.8%, yeast extract paste 0.5%, pH5.0.
Method: the 500ml capacity triangular flask loading amount of take is 100ml, and ground rice is weighed respectively to dress, inorganic salt unified preparation dissolve, then are sub-packed in each triangular flask, fill in tampon, with kraft paper, wrap up.0.11MPa, sterilizing 30min, cooling standby.
Two, inoculation, cultivation.
Get a test tube slant bacterial classification, with aseptic bamboo let, access in 3～5 liquid triangle shake-flask culture bases that (bottle number is depending on required grain weight, generally, in inoculum size 0.1%, it is 200mL that 200L seeding tank needs grain weight), in 30 ℃, the about 48h of 200rpm shaking table shaking culture, (incubation time is because of preservation time of test tube slant bacterial classification difference to some extent, general fresh preservation of bacteria strain required time is short, and longer bacterial classification of preservation time, incubation time is corresponding prolongation also), hand over workshop to check and accept standby.
S4, the preparation of seeding tank liquid seeds.
One, feed intake and disappear in fact.
1, nutrient solution formula: rice meal 7.5%, NaNO
30.4%, MgSO
40.1%, K
2hPO
40.1%, soya-bean oil 0.1%.
2, feeding method: after confirming that preparation work is errorless, can opening lid, more than adding water to bottom paddle, opens to stir and feed intake, continues to add water to constant volume after complete, builds cover, and air boosts, and whether inspection cover seepage, and whether visor is intact.Calculate beating time and be no less than 10min.
3, disappear in advance: when total steam reaches more than 0.2MPa, can carry out disinfection.First slowly open total steam valve, drive extrusion steam valve, the forcing valve cock of crack water drain valve and No. 1, No. 2 tank, closes two-way culture transferring valve; Drive sampling steam valve, close and sample into pot valve, crack sampling below valve; Close minute process filter turnover steam valve and air inlet one, two valves, open and sweep away steam valve, crack air enters pot valve; Carrying out pipeline disappears in advance.
4, preheating: open the preheating of jacket steam valve, open rear pressure and be no more than 0.2Mpa, open inoculation valve and cock, can opening vent valve, closes air robot under meter vent valve, when tank temperature rise to 80 ℃, closes jacket steam.
5, admission: close Yu Xiaoshige road exhaust valve, drive forcing valve, air enters pot valve, samples into pot valve.Require evenly, to prevent short circuit, in tank, stir well, tank exhaust is appropriate, controls total steam.
6, insulation: be slowly warmed up to 120～125 ℃, during tank pressure 0.1～0.4MPa, calculate soaking time 30min.During end, close each road admission valve and steam valve, dwindle steam discharge.
7, cooling: to close but hydrophone, open tap water backwater, lower the temperature, treat that tank pressure is down to 0.08MPa and stops stirring, open air and enter tank pressurize, be cooled to inoculation temp: 35～40 ℃ of inoculations.
8, inoculation environment dirt falls bacterium and removes: in process of cooling, fill with appropriate tap water in 20L atomizer, seeding tank surrounding environment water is sprayed, to remove in air the dirt such as dust, fall bacterium.
Two, inoculation.
1, inoculum size: 0.1%
2, inoculation method: clean visor mouth to be opened with 75% alcohol, in the situation that guaranteeing seeding tank malleation, unclamp seeding tank visor, with Alcohol Flame circle, seal visor mouth taking off visor simultaneously, then in flame ring, open triangle bottleneck, pour seed in triangular flask into seeding tank rapidly, remove flame ring simultaneously, cover visor, obturage.
3, culture condition and control: rotating speed: 240rpm, temperature: 30～32 ℃, tank pressure: 0.05～0.07MPa, air flow quantity: 8～10m
3/ hr, incubation time: approximately 48 hours.
S5, fermentation culture.
One, rice dipping: the packing bag that this process operations personnel should untie rice, rice is divided and is filled in puffed wheat bag, each puffed wheat bag approximately fills 20-25kg, with cotton thread, tightens sack, puts in puffed wheat pond and piles up neatly.
Two, drip washing: till water base the clarification of extremely flowing out with tap water drip washing rice, the object of cleaning is to wash out the foreign matters such as chaff shell that are attached to rice surface.After drip washing, running water pipe is spooled playback in order.
Three, drain: by standing for some time of the rice cleaning up, drain free water content, require to mention puffed wheat bag and flow down without the wire globule.
Four, steam rice: the rice draining is steamed to 2～3min at the temperature of 102 ℃.
Five, inoculation.
1, the preparation of bacterial classification: measure 16% inoculum size by rice and take bacterial classification, the Glacial acetic acid of amount of rice 1% before inoculation, mixes rear standby.
2, inoculation method: load weighted bacterial classification is evenly sprinkled upon on the rice of spreading for cooling, turns 3 times by hand, guarantee that bacterial classification fully mixes with material.The uniform bank of inoculation is overlayed to one end of woollen blanket under gauze, pile thick approximately 40～50cm, woollen blanket is added a cover on surface.
Six, beat heap.
1, high hot house is beaten heap
(1), the preparation of high hot house: before high hot house is used, (before puffed wheat) reply ground is cleared up, and checks that heating valve and power supply situation are to guarantee envrionment temperature in high hot house.When ambient temperature is during lower than 10 ℃, ground should overlay electric blanket.
(2), the management of high hot house: 1. high hot house is only played heap intensification for fermentation plant fermentation materials, forbids no-operation personnel to stay in high hot house.2. the high hot house waste of clean rooms in time of finishing using, fastens radiator valve, closes light switch, opens wide high hot house gate, and electric blanket dries rear timely playback.3. to irregularly to high hot house, carry out fumigation sterilizing.
(3), material after inoculation is installed with plastics bag, with cotton thread, tie sack, pile up in high hot house, note keeping room temperature between 25～40 ℃, and note observing room temperature and windrow temperature variations.While having partial material temperature to arrive 40 ℃, should be carried out a turning in high hot house, the material switch that temperature height is inhomogeneous is placed, when most of temperature of charge arrives more than 45 ℃, this batch materials can be gone to fermentation vat and grow flower.Under normal circumstances, windrow temperature rose to 45～48 ℃ at 15～24 hours.
2, Quchi is beaten heap: when room temperature is during higher than 25 ℃, after inoculation, material can be placed directly in and in Quchi, play heap cultivation.
(1), Da Dui place prepares: the woollen blanket of the sterilizing of tiling under the gauze of Quchi one end, when room temperature should overlay electric blanket during lower than 20 ℃ under woollen blanket, after completing, even up gauze.
(2), material after inoculation is transported in Quchi and is piled up with Turnover Box, pile thick approximately 40～50cm, woollen blanket is added a cover on surface, at material middle part, plugs thermometer.Note keeping room temperature between 25～30 ℃, and note observing windrow temperature variations.In transport process, note filling on the Turnover Box of thing and will add a cover the woollen blanket that sterilizing is good, the Turnover Box being finished down after material does not directly contact with ground yet.
Seven, stand heap is grown flower.
1, high hot house material stand heap is grown flower: in high hot house, most of temperature of charge arrives more than 45 ℃, while having partial material product temperature to reach 48 ℃, this batch materials can be gone to fermentation vat and again beat heap and grow flower.With transfer car(buggy), high hot house material is gone to bent room, on the ground in bent room, spread and clean the plastic cloth drying, material is placed on plastic cloth, do not untie tying rope, material in bag is rubbed loose, untie tying rope, be poured on Quchi one end and again beat heap, pile thick 30～40cm, when the centre of material product temperature rises to 45 ℃ again, full pond material is thoroughly turned one time, make thinner, pile thick maintenance 20～30cm, 38～42 ℃ of maintenance product temperature are full full pond, stand after approximately 3～5 hours, during adopt as far as possible and turn over song and guarantee temperature of charge by hand.On most of material during existing monascus bacterial plaque, can be by product temperature control at 35～38 ℃ until add a water.During growing flower, at least to guarantee at interval of 3～4 hours thorough stirrings once.
2, Quchi material stand heap is grown flower: when Quchi material centre product temperature rise to 48 ℃, full pond material is thoroughly turned one time, again beat heap, pile thick 20～30cm, when the centre of material product temperature rises to 45 ℃ again, full pond material is thoroughly turned one time to the full full Chi Dui in stand, 38～42 ℃ of maintenance product temperature approximately 3～5 hours, use less during this time gas blower air blast, turning over song guarantees temperature of charge by hand in employing as far as possible as far as possible.On most of material during existing monascus bacterial plaque, can be by product temperature control at 35～38 ℃ until add a water.During growing flower, at least to guarantee at interval of 3～4 hours thorough stirrings once.
Eight, add water.
1, add 1 water: growing flower neatly, (90% grain of rice surface white hypha evenly infiltrates, and the part grain of rice reddens, generally at stand, pile latter about 24 hours), this stage grain of rice starch is not also made full use of by monascus, monascus is in the high-speed rapid growth stage, and thermal value is larger, therefore, will repeatedly add on a small quantity 1 water, and amount of water is larger.
Water feeding method: Bian Jiashui, limit stirring, depending on loose, the dry wet situation of bent material, repeat above-mentioned steps, until material is moisture evenly after adding water, moisture is (4～October, moisture can remain between 43%～48%, and March November to next year, moisture can remain between 40%～45%) between 40%～48%.Requirement add water after 1 hour on moistening, the Quchi gauze of material and material surface without free-water.
Product temperature control: add the control of noting material product temperature after 1 water, operator to guarantee song go out in more than 80% material product temperature remain between 33 ℃-35 ℃.
2, add 2 water: the growing state depending on material is determined the time that adds two water, after adding 1 water general 4～October, within approximately 10 hours, need add two water, March November to next year will be according to the intensification situation of material, determine the time that adds 2 water, material water suction is fast, quick heating, within approximately 10 hours after adding 1 water, add 2 water, otherwise can within 24 hours after adding 1 water, add 2 water.Water feeding method is the same.After adding water, moisture controlled is at 43-50%, and product temperature control is 33 ℃ of left and right, and notices that the local product temperature of this stage material must not be lower than 30 ℃, and high temperature must not be over 34 ℃.
3, intermediary and later stages culturing process adds water and goes out pond and dries and within first 1 day, add water: intermediary and later stages culturing process adds water water feeding method and requires with 2 water; Go out pond dries and to add water the day before yesterday: go out pond whether add water the day before yesterday will be according to water content detection result on the same day, if moisture, more than 43%, need not add water, otherwise suitably moisturizing, after moisturizing, moisture requirement is no more than 48%, product temperature control is the same.
S6, go out pond and dry.
S7, pulverizing.
S8, packing.
S9, check.
S10, finished product warehouse-in.
Positively effect of the present invention: 1) to take Monascus fulginosus (Monascus ruber) etc. be starting strain to fermentation red colouring agent for food, also used as a Chinese medicine, by many bacterial classifications assortment of genes technology, filter out high saccharogenic power, the composite bacteria of high esterifying power and generation aspartic protease and reddish yellow pigment.
2) this composite bacteria produces aspartic protease, saccharifying enzyme and Esterified Enzyme, can produce alcohol and carbonic acid gas by decomposition glucose, the alcohol that a part generates, and a part is oxidized to organic acid, and a part is combined to ester with amino acid and organic acid etc.Therefore after using, ester is aromatic strongly fragrant.This composite bacteria is to also there being good enzyme to live in high salt, peracid situation.
3) fermentation red colouring agent for food, also used as a Chinese medicine be pure fermenting organism viable bacteria goods, by koji ferment generation abundant enzyme be.The saccharification flavoring sort of quyi can resist higher osmotic pressure, can be in pH value below 7.0, and 48 ℃ of tolerable temperatures, the suitableeest 30 ℃～35 ℃, salinity 22B ē degree is used below.
4) the fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification compound sort of quyi of thing fermentative production of making a living, by the koji abundant enzyme of generation that ferment is.The infull defect of single culture koji enzyme system can be made up, the vigor of the enzyme systems such as bent proteolytic enzyme, saccharifying enzyme, Esterified Enzyme can be obviously improved into.Improve protein utilization and the yield rate of raw material.
5) fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification has stronger product look ability, during the fermentation, produces haematochrome and yellow pigment, improves Red Index and the yellowness index of product, makes that product color is bright-coloured, glow.
6) fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification can resist higher osmotic pressure, can be in pH value below 7.0, and 48 ℃ of tolerable temperatures, 30 ℃～35 ℃ of optimum temperutures, 22 ° of B é of salinity are used below.
7) application art is simple, in soy sauce and jam product koji, only need in proportion saccharification flavoring yeast be mixed with aspergillus oryzae, more evenly add the fermentation of material koji.In Vinegar Fermentation process, directly add.
8) fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification is comprised of a plurality of monascuses such as fermentation red colouring agent for food, also used as a Chinese medicine, esterified red yeast and product proteolytic enzyme red colouring agent for food, also used as a Chinese medicine.
9) application of fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification in fermented sauce and jam product adopts mixed culture technology.
Realization in order to demonstrate the invention, has described above-mentioned embodiment.But other variations of the present invention and modification, be apparent for those skilled in the art, any modification/variation within the scope of essence disclosed in this invention and fundamental principle or imitation conversion all belong to claim protection domain of the present invention.
Claims (6)
1. a fermentation red colouring agent for food, also used as a Chinese medicine bacterial classification, is characterized in that, this bacterial classification is preserved in Wuhan, Hubei loujia hill belongs Wuhan University Chinese Typical Representative culture collection center on February 23rd, 2011, and preserving number is: CCTCC NO:M2011047.
2. the ferment production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification, is characterized in that, comprises the following steps:
Step 1, vacuum freezing pipe at 4 ℃, the preservation of normal temperature refrigerator;
Step 2, the preparation of test tube slant bacterial classification:
One, conditioned media: peptone 2%, Zulkovsky starch 2%, Fructus Hordei Germinatus soak powder 2%, (NH
4)
2sO
40.1%, MgSO
40.1%, K
2hPO
40.1%, agar 2%, natural pH; With 18 * 180 (mm) test tube, every pipe dress substratum 7～10ml (substratum is hot melt fully), with tampon, seal, 10 test tube bag a bundles, vertically be put in iron wire frame, 0.11MPa sterilizing 30min, is cooled to 50～60 ℃, incline inclined-plane processed (require slant medium length be test tube 3/4), after condensation, wrap up bundled; Cultivate 48hr, on inspection without miscellaneous bacteria, can use for 30 ℃;
Two, inoculation culture:
1, vacuum freezing ampoul tube turns test tube slant bacterial classification: with aseptic capillary pipet, one pipe stroke-physiological saline solution is added in ampoul tube, stir evenly, bacterium liquid is sucked to capillary pipet, in access slant tube; Cultivate 3～5 days for 30 ℃, switching activated for 2 to 3 generations, cultivated 8～10 days, put 4 ℃, refrigerator and save backup for 30 ℃;
2, slant tube strain transfer: get a slant tube kind, wipe examination totally with 75% spirituous solution, access 20～30 test tube slants with aseptic bamboo let in super clean bench, on spirit lamp flame; It is identical that cultivation and store method and described vacuum freezing ampoul tube turn test tube slant bacterial classification;
Step 3, the preparation of liquid triangle shaking flask bacterial classification:
One, substratum: glucose 4%, NaNO
30.2%, MgSO
40.1%, extractum carnis 0.3%, peptone 0.8%, yeast extract paste 0.5%, pH5.0; The 500ml capacity triangular flask loading amount of take is 100ml, and ground rice is weighed respectively to dress, and inorganic salt unified preparation dissolve, then are sub-packed in each triangular flask, fill in tampon, with kraft paper, wrap up.0.11MPa, sterilizing 30min, cooling standby;
Two, inoculation, cultivation: get a test tube slant bacterial classification, with aseptic bamboo let, access in 3～5 liquid triangle shake-flask culture bases, in 30 ℃, the about 48h of 200rpm shaking table shaking culture, hand over workshop to check and accept standby;
Step 4, the preparation of seeding tank liquid seeds:
1, nutrient solution formula: rice meal 7.5%, NaNO
30.4%, MgSO
40.1%, K
2hPO
40.1%, soya-bean oil 0.1%; With 75% alcohol, clean visor mouth to be opened, in the situation that guaranteeing seeding tank malleation, unclamp seeding tank visor, with Alcohol Flame circle, seal visor mouth taking off visor simultaneously, then in flame ring, open triangle bottleneck, pour seed in triangular flask into seeding tank rapidly, remove flame ring simultaneously, cover visor, obturage;
2, culture condition and control: rotating speed: 240rpm, temperature: 30～32 ℃, tank pressure: 0.05～0.07MPa, air flow quantity: 8～10m
3/ hr, incubation time: approximately 48 hours;
Step 5, fermentation culture:
Rice dipping, drip washing, drain, steam rice: the rice draining is steamed to 2～3min at the temperature of 102 ℃;
The preparation of bacterial classification: measure 16% inoculum size by rice and take bacterial classification, the Glacial acetic acid of amount of rice 1% before inoculation, mixes rear standby;
Load weighted bacterial classification is evenly sprinkled upon on the rice of spreading for cooling, turns 3 times by hand, guarantee that bacterial classification fully mixes with material.The uniform bank of inoculation is overlayed to one end of woollen blanket under gauze, pile the thick 40～50cm of being, woollen blanket is added a cover on surface;
Step 6, Da Dui: be divided into that high hot house is beaten heap and Quchi is beaten heap, room temperature during lower than 20 ℃ high hot house beat heap; Room temperature during higher than 20 ℃ Quchi beat heap, pile thick 35～40cm, surface, coated with the felt insulation of sterilizing, keeps 20～30 ℃ of room temperatures, 15～20hr;
Step 7, stand heap are grown flower: comprise that high hot house material stand heap is grown flower and Quchi material stand heap is grown flower, 45～48 ℃ of stand heaps, keep 20～30 ℃ of room temperatures, and product temperature maintains 32～35 ℃, 20hr;
Step 8, Jia Shui: keep material moisture approximately 35%～55%, room temperature, at 20～25 ℃, is cultivated 4 days;
Step 9, go out pond and dry.
3. the ferment production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification, is characterized in that, described high hot house is beaten heap and referred to: material after inoculation is installed with plastics bag, with cotton thread, tie sack, pile up in high hot house, notice that maintenance room temperature is between 25～35 ℃; While having partial material temperature to arrive 40 ℃, should be carried out a turning in high hot house, the material switch that temperature height is inhomogeneous is placed, when most of temperature of charge arrives more than 45 ℃, this batch materials be gone to fermentation vat and grow flower; Under normal circumstances, windrow temperature rose to 45～48 ℃ at 15～24 hours.
4. the ferment production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification, is characterized in that, described Quchi is beaten heap and referred to: when room temperature is during higher than 25 ℃, after inoculation, material can be placed directly in and in Quchi, play heap cultivation; Material after inoculation is transported in Quchi and is piled up with Turnover Box, pile thick approximately 40～50cm, woollen blanket is added a cover on surface, at material middle part, plugs thermometer.Note keeping room temperature between 25～30 ℃, and note observing windrow temperature variations.In transport process, note filling on the Turnover Box of thing and will add a cover the woollen blanket that sterilizing is good, the Turnover Box being finished down after material does not directly contact with ground yet.
5. the production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification of fermenting, it is characterized in that, described high hot house material stand heap is grown flower and is referred to: in high hot house, most of temperature of charge arrives more than 45 ℃, while having partial material product temperature to reach 48 ℃, this batch materials is gone to fermentation vat and again beat heap and grow flower; With transfer car(buggy), high hot house material is gone to bent room, on the ground in bent room, spread and clean the plastic cloth drying, material is placed on plastic cloth, do not untie tying rope, material in bag is rubbed loose, untie tying rope, be poured on Quchi one end and again beat heap, pile thick 30～40cm, when the centre of material product temperature rises to 45 ℃ again, full pond material is thoroughly turned one time, make thinner, pile thick maintenance 20～30cm, 38～42 ℃ of maintenance product temperature are full full pond, stand after approximately 3～5 hours, during adopt as far as possible and turn over song and guarantee temperature of charge by hand.On most of material during existing monascus bacterial plaque, can be by product temperature control at 35～38 ℃ until add a water; During growing flower, at least to guarantee at interval of 3～4 hours thorough stirrings once.
6. the production technique of red colouring agent for food, also used as a Chinese medicine bacterial classification of fermenting, it is characterized in that, described Quchi material stand heap is grown flower and is referred to: when Quchi material centre product temperature rise to 48 ℃, full pond material is thoroughly turned one time, again beat heap, pile thick 20～30cm, when the centre of material product temperature rises to 45 ℃ again, full pond material is thoroughly turned one time, the full full Chi Dui in stand, 38～42 ℃ of maintenance product temperature approximately 3～5 hours, during use less gas blower air blast as far as possible; On most of material during existing monascus bacterial plaque, by product temperature control at 35～38 ℃ until add a water.During growing flower, at least to guarantee at interval of 3～4 hours thorough stirrings once.
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| CN105754797A (en) * | 2016-05-09 | 2016-07-13 | 福建师范大学 | Preparation method of monascus coated with yellow spores |
| CN106937736A (en) * | 2017-03-08 | 2017-07-11 | 湖北工业大学 | A kind of soy sauce and preparation method thereof |
| CN108165497A (en) * | 2017-12-22 | 2018-06-15 | 吕玲 | A kind of Monascus Strains breeding method of high yield Mo Nakelin K |
| CN110272830A (en) * | 2019-07-08 | 2019-09-24 | 武汉佳成生物制品有限公司 | The Monascus fulginosus bacterium and its application method of a kind of high-yield glucoamylase and high germination rate |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105349438A (en) * | 2015-12-15 | 2016-02-24 | 武汉佳成生物制品有限公司 | Functional red yeast rice strain and production process thereof |
| CN105349437A (en) * | 2015-12-15 | 2016-02-24 | 武汉佳成生物制品有限公司 | High color value monascorubin strain and production process thereof |
| CN105754797A (en) * | 2016-05-09 | 2016-07-13 | 福建师范大学 | Preparation method of monascus coated with yellow spores |
| CN105754797B (en) * | 2016-05-09 | 2018-07-17 | 福建师范大学 | A kind of preparation method of yellow clothes red yeast rice |
| CN106937736A (en) * | 2017-03-08 | 2017-07-11 | 湖北工业大学 | A kind of soy sauce and preparation method thereof |
| CN108165497A (en) * | 2017-12-22 | 2018-06-15 | 吕玲 | A kind of Monascus Strains breeding method of high yield Mo Nakelin K |
| CN110317734A (en) * | 2019-07-02 | 2019-10-11 | 江苏今世缘酒业股份有限公司 | A kind of monascus and its isolated culture method and the application of high-yield glucoamylase, Esterified Enzyme and protease |
| CN110272830A (en) * | 2019-07-08 | 2019-09-24 | 武汉佳成生物制品有限公司 | The Monascus fulginosus bacterium and its application method of a kind of high-yield glucoamylase and high germination rate |
| CN110452822A (en) * | 2019-08-22 | 2019-11-15 | 武汉佳成生物制品有限公司 | A kind of compound wine brewing song and its brewing method using vinasse fermentation wine brewing |
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