CN104004677B - A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation - Google Patents
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation Download PDFInfo
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Abstract
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation, processes including the collection of alcohol industry yellow fluid, the collecting and prepare of Bacillus licheniformis cultivation in alcohol industry yellow fluid and Bacillus licheniformis.The method utilizing alcohol industry yellow fluid to produce Bacillus licheniformis provided by the present invention, alcohol industry yellow fluid can be made full use of, reduce the processing cost of yellow fluid, optimize the comprehensive utilization of yellow fluid, improve the ecological environment, existing Bacillus licheniformis preparation can be expanded again and produce the source of Middle nutrition substrate, reduce production cost, expand the range of application of Bacillus licheniformis.
Description
Technical field
The present invention relates to field of comprehensive utilization and the field of microbial fermentation of alcohol industry yellow fluid, especially with ethanol
Yellow fluid in industry carries out fermentable.
Background technology
Alcohol industry is one important energy industry of China, but can produce huge by-product in alcohol production process
Thing, wherein the solid constituent such as distiller grains can continue to produce the feedstuffs such as DDGS and be used again together with part yellow fluid, and exhausted
Most yellow fluid is the most well utilized.Yellow fluid be produce discharge during ethanol rich in multiple nutritional components
Liquid substance, in yellow fluid containing abundant organic acid carboxylic acid contents such as (especially most) lactic acid, butanoic acid, caproic acid, acetic acid, alcohol,
Aldehyde, ester etc. are in fragrance matter, simultaneously possibly together with aminoacid, saccharide, ethanol, humus, yeast thalline autolysate, microbial cells
And living cells etc., nutritious due to it, it is directly discharged in environment the pollution often causing serious body eutrophication,
Process to this end, the waste water of national regulation ethanol enterprise discharge has to pass through early stage, and can discharge after reaching 4 grades of water quality standards, this
Sample one, in addition to the nutritional labeling in yellow fluid is wasted, ethanol enterprise often increases the process for yellow fluid of the more expense,
And, major part brewery of current China is all to use it for mixing grain return cellar for storing things fermentation or support cellar for storing things, cultivate artificial cellar for storing things to the process of yellow fluid
The aspects such as mud, not only can not efficent use of resources, and cannot fundamentally solve ferment yellow fluid final outlet problem.Therefore
How to comprehensively utilize yellow fluid resource, how can turn waste into wealth, the outlet problem thoroughly solving yellow fluid is particularly important, if
Yellow fluid can be turned waste into wealth, fully utilize, the cost reducing enterprise be put into, also complies with the need of improvement of the ecological environment simultaneously
Want.
Bacillus licheniformis is Gram-positive bacillus, cell size.Bacillus licheniformis
Cellular morphology and arrangement are in shaft-like, Dan Sheng, and its spore form is to produce nearly middle raw ellipticity spore, and sporangiocyst slightly expands.Lichens
After bacillus cereus enters intestinal as active bacteria formulation, the pathogenic bacterium such as staphylococcus, yeast-like fungus there are is antagonism, and to bifid
The probioticss such as bacillus, lactobacillus, bacteroid, digestibility streptococcus have facilitation, thus reach to regulate microbial population of animal intestinal tract
Balance, improves the effect of enteric microorganism environment；Can the growth promoter of stimulating animal immune organ, activated lymphocyte, carry
High immunoglobulinlg and antibody horizontal, strengthen cellular immunization and humoral immune function, improves immunity of organisms；It addition, suitably
Under the conditions of, it is possible to the enzymes such as synthetic proteins enzyme, lipase, amylase and cellulase, jointly play a role with endogenous enzymes, promote
Digesting and assimilating of nutrient.Based on above feature, current Bacillus licheniformis is at row such as medicine, health care, food, biological feedstuff productions
Industry is widely applied.But, the many employings of production method of current Bacillus licheniformis preparation artificially add a large amount of high-quality battalion
Support material produce as raw material, its production cost is higher, and then limits the application of Bacillus licheniformis, and increase with
Bacillus licheniformis is the cost of the Related product of raw material.
Due in alcohol industry yellow fluid rich in the nutrient substance of multiple soluble state, if it is possible to utilize alcohol industry
Yellow fluid carries out the production of Bacillus licheniformis, can solve the problem of complex utilization of alcohol industry yellow fluid, turn waste into wealth, and can drop again
The production cost of low current Bacillus licheniformis preparation, expands the range of application of Bacillus licheniformis further.
Summary of the invention
It is an object of the invention to provide a kind of production method utilizing alcohol industry yellow fluid to carry out Bacillus licheniformis preparation,
On the one hand reduce link and expense that in alcohol industry, yellow fluid processes, improve the comprehensive utilization ratio of alcohol industry yellow fluid, the opposing party
Face reduces the industrial cost of Bacillus licheniformis preparation.
The present invention solves above-mentioned technical problem and the technical scheme is that one utilizes alcohol industry yellow fluid to produce lichens bud
The method of spore bacillus preparation, it specifically comprises the following steps that
(1) directly collect alcohol industry yellow fluid from the sheet frame of filter press, put into rapidly in the container of sealing, then take
Sample carries out the mensuration of the nutrient composition contents such as protein, cellulose, aminoacid, organic acid, mineral element, and according to practical measurement
Value and the nutritional need of Bacillus licheniformis growth and breeding, carry out nutritional labeling and supplement；
(2) the alcohol industry yellow fluid sterilizing under the conditions of 121 DEG C in high-pressure sterilizing pot after supplementing the nutrients in step (1)
15 ~ 20min, and cool down, the alcohol industry yellow fluid pH after sterilizing is 3.2 ~ 3.5, is not suitable for Bacillus licheniformis raw
Long breeding, then utilizes sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5；
(3) Bacillus licheniformis is inoculated in beef extract-peptone fluid medium, cultivates to steady production phase, detection
Viable count is not less than 1 × 109 Cfu/mL can be used as seed bacterium solution, standby, wherein, and described beef extract-peptone liquid culture
The proportioning of base is: Carnis Bovis seu Bubali cream 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5；
(4) by seed bacterium solution obtained by step (3) according to 6% ~ 8%(v/v) ratio be inoculated in after step (2) processes
Alcohol industry yellow fluid in, and at a temperature of 36 DEG C ~ 36.5 DEG C, be placed on the constant temperature oscillator of 200 ~ 220r/min cultivation 16
~ 18h, obtains Bacillus licheniformis liquid, standby；
(5) adding calcium chloride in the Bacillus licheniformis liquid obtained in step (4), its amount added is according to weighing body
Long-pending is 0.05% ~ 0.08% than (w/v), then heats 10min ~ 20min in 60 DEG C of water-baths, then proceeds to heat in 80 DEG C of water-baths
5min ~ 10min, obtains the bacterium solution containing brood cell, the Bacillus licheniformis spore forming rate 89.1% ~ 92.6% that the method processes；
(6) by bacterium solution Large Copacity centrifuge containing spore in step (5) in 3500 ~ 4000r/min be centrifuged 10 ~
15min, abandoning supernatant, it is thus achieved that Bacillus licheniformis solids；
(7) according to Bacillus licheniformis solids and the weight ratio of carrier 1:4 ~ 6 of step (6) gained, calcium carbonate is added
Powder carrier, mix homogeneously, make Bacillus licheniformis solid mixture；
(8) by the Bacillus licheniformis solid mixture of step (7) gained aseptic, dry under the conditions of 60 DEG C ~ 65 DEG C, grind
Wearing into dry powder, in dry powder, effective Bacillus licheniformis number is 20 × 109~30×109Individual/g, effective Bacillus licheniformis number refers to
It is Bacillus licheniformis viable count and the Bacillus licheniformis number sum forming spore；
(9) in the Bacillus licheniformis dry powder obtained by step (8) according to weight ratio add 4% lactose, 20% can
Soluble starch, as excipient, mix homogeneously, makes Bacillus licheniformis preparation, wherein effective Bacillus licheniformis number >=5,000,000,000
Individual/gram, then with laminated film to lichens bacillus preparation, it being vacuum-packed, vacuum is 0.085 ~ 0.095Mpa.
Bacillus licheniformis used by the present invention is purchased from Sang Ge bio tech ltd, Shanghai, is preserved in Unite States Standard raw
Article collecting center, its deposit number is ATCC 11946, and the purchase date is on November 20th, 2013.
The method have the advantages that
First, compared with the raw material of current Bacillus licheniformis preparation production and nutrient matrix, the present invention uses ethanol work
Industry yellow fluid, as cultivating raw material and substrate, takes full advantage of alcohol industry waste water, improves the comprehensive utilization of alcohol industry yellow fluid
It is worth, improves ecological environment, expand Bacillus licheniformis preparation and produce the source of desired nutritional substrate, reduce lichens bud
The production cost of spore bacillus；
Second, the nutrient substance in the cultivation of Bacillus licheniformis, alcohol industry yellow fluid is abundant by Bacillus licheniformis
Utilizing, content effectively reduces, and basically reaches the discharge standard of alcohol waste water, does not results in the eutrophication of water body, and saves
The link of business processes alcohol industry yellow fluid and relevant put into, reduces the production cost of relevant enterprise；
3rd, the present invention is directly to gather alcohol industry yellow fluid from the sheet frame of filter press, this is because ethanol work
The bleeding point of industry yellow fluid is the most important, and the yellow fluid nutritional labeling directly collected from the sheet frame of filter press is collected compared with discharge outlet
Yellow fluid rich in nutrition content, is more suitable for as culture matrix；
4th, the present invention is on the premise of measuring alcohol industry yellow fluid nutrient content, according to measurement result targetedly
Yellow fluid is carried out nutritional supplementation, to be more suitable for growth and the breeding of lichens bacillus cereus, further increases lichens spore bar
Bacterium yield in alcohol industry yellow fluid；
5th, for the distinctive chemical composition of alcohol industry yellow fluid, carry out suitable pre-treatment and part chemical property
Change so that it is be more suitable for growth and the breeding of Bacillus licheniformis, improve the yield of Bacillus licheniformis；
6th, for peculiar chemical composition and the character of alcohol industry yellow fluid, it is provided that suitable being used for cultivates lichens bud
The operating parameter of spore bacillus, including Bacillus licheniformis inoculum concentration, cultivation temperature, shaking speed and incubation time etc.；
7th, after Bacillus licheniformis is cultivated and terminates, utilize that the present invention provides collect and preparation method can make ground
Effective bacterium number >=5,000,000,000/gram of clothing bacillus preparation, spore forming rate reaches 89.1% ~ 92.6%.
Therefore, the method utilizing alcohol industry yellow fluid to produce Bacillus licheniformis provided by the present invention, can be fully sharp
Use alcohol industry yellow fluid, reduce the processing cost of yellow fluid, optimize the comprehensive utilization of yellow fluid, improve the ecological environment, can expand again existing
There is Bacillus licheniformis preparation to produce the source of Middle nutrition substrate, reduce production cost, expand the application model of Bacillus licheniformis
Enclose.
Accompanying drawing explanation
Fig. 1 is the content of various nutritional labelings in alcohol industry yellow fluid.
Fig. 2 is the different cultivation temperature impacts on effective Bacillus licheniformis number.
Fig. 3 is the different incubation time impacts on effective Bacillus licheniformis number.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1: a kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation, this method uses
Semen Maydis is the yellow fluid of raw material production ethanol, specifically comprises the following steps that
(1) directly collect alcohol industry yellow fluid from the sheet frame of filter press, put into rapidly in the container of sealing, then take
Sample carries out the mensuration of the nutrient composition contents such as protein, cellulose, aminoacid, organic acid, mineral element, obtains alcohol industry yellow
The content of various nutritional labelings in water, measurement result is shown in accompanying drawing 1, and according to practical measurement value and Bacillus licheniformis growth and breeding
Nutritional need, carry out nutritional labeling and supplement, concrete nutritional labeling supplements kind and quantity is: every liter of above-mentioned alcohol industry is yellow
Water adds carbamide 5g, glucose 2g and sodium chloride 2g；
(2) the alcohol industry yellow fluid sterilizing under the conditions of 121 DEG C in high-pressure sterilizing pot after supplementing the nutrients in step (1)
15 ~ 20min, and cool down, then utilize sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5；
(3) Bacillus licheniformis is inoculated in beef extract-peptone fluid medium, cultivates to steady production phase, detection
Viable count is not less than 1 × 109 Cfu/mL can be used as seed bacterium solution, standby, wherein, and described beef extract-peptone liquid culture
The proportioning of base is: Carnis Bovis seu Bubali cream 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5；
(4) by seed bacterium solution obtained by step (3) according to 6%(v/v) ratio be inoculated in after step (2) processes
In alcohol industry yellow fluid, and at a temperature of 36 DEG C, it is placed on the constant temperature oscillator of 200 ~ 220r/min cultivation 16h, obtains ground
Clothing bacillus cereus bacterium solution, standby；
(5) adding calcium chloride in the Bacillus licheniformis liquid obtained in step (4), its amount added is according to weighing body
Long-pending is 0.05% than (w/v), then heats 10min in 60 DEG C of water-baths, then proceeds to heat in 80 DEG C of water-baths 5min, is contained
The bacterium solution of brood cell；
(6) by bacterium solution Large Copacity centrifuge containing spore in step (5) in 3500 ~ 4000r/min be centrifuged 10 ~
15min, abandoning supernatant, it is thus achieved that Bacillus licheniformis solids；
(7) according to Bacillus licheniformis solids and the weight ratio of carrier 1:4 of step (6) gained, Paris white is added
Carrier, mix homogeneously, make Bacillus licheniformis solid mixture；
(8) by the Bacillus licheniformis solid mixture of step (7) gained aseptic, dry under the conditions of 60 DEG C ~ 65 DEG C, grind
Wear into dry powder；
(9) in the Bacillus licheniformis dry powder that step (8) obtains, the lactose of 4%, 20% solvable are added according to weight ratio
Property starch, as excipient, mix homogeneously, makes Bacillus licheniformis preparation, then with laminated film to Bacillus licheniformis system
Agent is vacuum-packed, and vacuum is 0.085 ~ 0.095Mpa.
After measured, Bacillus licheniformis effective bacterium number in the method gained Bacillus licheniformis preparation in the present embodiment is used
For (41.5 ± 5.3) × 109。
Embodiment 2: a kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation, this method uses
Semen Maydis is the yellow fluid of raw material production ethanol, specifically comprises the following steps that
(1) directly collect alcohol industry yellow fluid from the sheet frame of filter press, put into rapidly in the container of sealing, then take
Sample carries out the mensuration of the nutrient composition contents such as protein, cellulose, aminoacid, organic acid, mineral element, obtains alcohol industry yellow
The content of various nutritional labelings in water, measurement result is shown in accompanying drawing 1, and according to practical measurement value and Bacillus licheniformis growth and breeding
Nutritional need, carry out nutritional labeling and supplement, concrete nutritional labeling supplements kind and quantity is: every liter of above-mentioned alcohol industry is yellow
Water adds carbamide 5g, glucose 2g, sodium chloride 2g；
(2) the alcohol industry yellow fluid sterilizing under the conditions of 121 DEG C in high-pressure sterilizing pot after supplementing the nutrients in step (1)
15 ~ 20min, cooling, then utilize sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5；
(3) Bacillus licheniformis is inoculated in beef extract-peptone fluid medium, cultivates to steady production phase, detection
Viable count is not less than 1 × 109 Cfu/mL can be used as seed bacterium solution, standby, wherein, and described beef extract-peptone liquid culture
The proportioning of base is: Carnis Bovis seu Bubali cream 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5；
(4) by seed bacterium solution obtained by step (3) according to 8%(v/v) ratio be inoculated in after step (2) processes
In alcohol industry yellow fluid, and at a temperature of 36.5 DEG C, it is placed on the constant temperature oscillator of 200 ~ 220r/min cultivation 18h, obtains
Bacillus licheniformis liquid, standby；
(5) adding calcium chloride in the Bacillus licheniformis liquid obtained in step (4), its amount added is according to weighing body
Long-pending is 0.08% than (w/v), then heats 20min in 60 DEG C of water-baths, then proceeds to heat in 80 DEG C of water-baths 10min, is contained
The bacterium solution of brood cell；
(6) by bacterium solution Large Copacity centrifuge containing spore in step (5) in 3500 ~ 4000r/min be centrifuged 10 ~
15min, abandoning supernatant, it is thus achieved that Bacillus licheniformis solids；
(7) according to Bacillus licheniformis solids and the weight ratio of carrier 1:6 of step (6) gained, Paris white is added,
Mix homogeneously, makes Bacillus licheniformis solid mixture；
(8) by the Bacillus licheniformis solid mixture of step (7) gained aseptic, dry under the conditions of 60 DEG C ~ 65 DEG C, grind
Wear into dry powder；
(9) using in above-mentioned Bacillus licheniformis dry powder according to weight ratio add 4% lactose, 20% soluble starch as
Excipient, mix homogeneously, make Bacillus licheniformis preparation, then with laminated film, lichens bacillus preparation is carried out vacuum
Packaging, vacuum is 0.085 ~ 0.095Mpa.
After measured, Bacillus licheniformis effective bacterium number in the method gained Bacillus licheniformis preparation in the present embodiment is used
For (48.2 ± 4.3) × 109。
Embodiment 3: a kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation, this method uses
Semen Maydis is the yellow fluid of raw material production ethanol, specifically comprises the following steps that
(1) directly collect alcohol industry yellow fluid from the sheet frame of filter press, put into rapidly in the container of sealing, then take
Sample carries out the mensuration of the nutrient composition contents such as protein, cellulose, aminoacid, organic acid, mineral element, obtains alcohol industry yellow
The content of various nutritional labelings in water, measurement result is shown in accompanying drawing 1, and according to practical measurement value and Bacillus licheniformis growth and breeding
Nutritional need, carry out nutritional labeling and supplement, concrete nutritional labeling supplements kind and quantity is: every liter of above-mentioned alcohol industry is yellow
Water adds carbamide 5g, glucose 2g, sodium chloride 2g；
(2) the alcohol industry yellow fluid sterilizing under the conditions of 121 DEG C in high-pressure sterilizing pot after supplementing the nutrients in step (1)
15 ~ 20min, cooling, then utilize sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5；
(3) Bacillus licheniformis is inoculated in beef extract-peptone fluid medium, cultivates to steady production phase, detection
Viable count is not less than 1 × 109 Cfu/mL can be used as seed bacterium solution, standby, wherein, and described beef extract-peptone liquid culture
The proportioning of base is: Carnis Bovis seu Bubali cream 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5；
(4) by seed bacterium solution obtained by step (3) according to 7%(v/v) ratio ethanol after processing through step (2)
In industry yellow fluid, and at a temperature of 36.2 DEG C, it is placed on the constant temperature oscillator of 200 ~ 220r/min cultivation 17h, obtains lichens
Bacillus cereus bacterium solution, standby；
(5) adding calcium chloride in the Bacillus licheniformis liquid obtained in step (4), its amount added is according to weighing body
Long-pending is 0.07% than (w/v), then heats 15min in 60 DEG C of water-baths, then proceeds to heat in 80 DEG C of water-baths 8min, is contained
The bacterium solution of brood cell；
(6) by bacterium solution Large Copacity centrifuge containing spore in step (5) in 3500 ~ 4000r/min be centrifuged 10 ~
15min, abandoning supernatant, it is thus achieved that Bacillus licheniformis solids；
(7) according to Bacillus licheniformis solids and the weight ratio of carrier 1:5 of step (6) gained, Paris white is added,
Mix homogeneously, makes Bacillus licheniformis solid mixture；
(8) by the Bacillus licheniformis solid mixture of step (7) gained aseptic, dry under the conditions of 60 DEG C ~ 65 DEG C, grind
Wear into dry powder；
(9) using in above-mentioned Bacillus licheniformis dry powder according to weight ratio add 4% lactose, 20% soluble starch as
Excipient, mix homogeneously, make Bacillus licheniformis preparation, then with laminated film, lichens bacillus preparation is carried out vacuum
Packaging, vacuum is 0.085 ~ 0.095Mpa.
After measured, Bacillus licheniformis effective bacterium number in the method gained Bacillus licheniformis preparation in the present embodiment is used
For (45.6 ± 6.4) × 109。
Below in conjunction with test example, the present invention is further illustrated.
Test example 1: inquire into and use method provided by the present invention to carry out Bacillus licheniformis cultivation, different cultivation temperature pair
The impact of effective bacterium number in final Bacillus licheniformis finished product, and find out optimum cultivation temperature.
In the cultivation of microorganism, the impact of temperature is most important, directly influences the success or not of microorganism culturing, with
And whether final colony counts meets the requirements.The method provided according to the present invention, uses single factor design, will cultivate temperature
Degree is respectively set to the different temperatures gradient of 35 DEG C, 35.5 DEG C, 36 DEG C, 36.5 DEG C, 37 DEG C, 37.5 DEG C, 39 DEG C and 40 DEG C, carries out
The cultivation of Bacillus licheniformis, result of the test is shown in accompanying drawing 2.
By time in accompanying drawing 2 it can be seen that carry out the cultivation of Bacillus licheniformis according to the method for the present invention, work as cultivation temperature
When 36 DEG C to 36.5 DEG C, in Bacillus licheniformis preparation, effective bacterium number is higher, and when especially 36.5 DEG C, effective bacterium number is
Height, therefore, 36.5 DEG C is the optimum cultivation temperature using the method for the present invention to cultivate Bacillus licheniformis.
Test example 2: inquire into and use method provided by the present invention to carry out Bacillus licheniformis cultivation, different incubation times pair
The impact of effective bacterium number in final lichens bacillus preparation, and find out optimum incubation time.
In microorganism culturing, the time too short bacterium colony that may can not get effective quantity, overlong time may be by miscellaneous bacteria
Polluting, therefore, incubation time is most important.According to the method for the present invention, use single factor design, by incubation time respectively
Being set to 12h, 16h, 18h, 24h, 36h and 48h, carry out the cultivation of Bacillus licheniformis, result of the test is shown in accompanying drawing 3.
Time from accompanying drawing 3 it can be seen that incubation time is 16 ~ 18h, in Bacillus licheniformis preparation, effective bacterium quantity is relatively
Many, especially when incubation time is 18h, Bacillus licheniformis effective bacterium number reaches the highest, therefore according to the method for the present invention,
Suitable incubation time is 18h.
Claims (6)
1. one kind utilizes the method that alcohol industry yellow fluid produces lichens bacillus preparation, it is characterised in that: comprise the following steps:
(1) directly collecting alcohol industry yellow fluid from the sheet frame of filter press, put in the container of sealing, sampling carries out nutrition one-tenth
Point mensuration, and according to measured value, collected alcohol industry yellow fluid carried out nutritional labeling and supplements, standby；
(2) the alcohol industry yellow fluid supplemented through nutritional labeling in step (1) is carried out sterilizing, pH regulator to 7.2 ~ 7.5, standby
With；
(3) being cultivated in beef extract-peptone fluid medium by Bacillus licheniformis to the steady production phase, detection viable count is not
Less than 1 × 109 Cfu/mL is as seed bacterium solution, standby；
(4) seed bacterium solution obtained by step (3) is inoculated according to the ratio that volume ratio is 6% ~ 8% processes institute through step (2)
In the alcohol industry yellow fluid obtained, and under the cultivation temperature of 36 DEG C ~ 36.5 DEG C, constant temperature oscillator is cultivated 16h ~ 18h, obtains
Bacillus licheniformis liquid, standby；
(5) adding calcium chloride in the Bacillus licheniformis liquid obtained in step (4), its amount added is according to w/v
It is 0.05% ~ 0.08%, in 60 DEG C of water-baths, then heats 15min ~ 20min, then proceed to 80 DEG C of water-baths are heated 5min ~ 10min,
Obtain the bacterium solution containing spore；
(6) it is centrifuged processing by the bacterium solution centrifuge containing spore in step (5), abandoning supernatant, it is thus achieved that lichens spore
Bacillus solids；
(7) according to Bacillus licheniformis solids and the part by weight of carrier 1:4 ~ 6 of step (6) gained, Paris white is added
Carrier, and mix homogeneously, make Bacillus licheniformis solid mixture；
(8) by the Bacillus licheniformis solid mixture that obtains in step (7) aseptically, in 60 DEG C ~ 65 DEG C drying, grind
Wear into dry powder；
(9) in the dry powder that step (8) obtains, add excipient, mix homogeneously, make Bacillus licheniformis preparation, and vacuum packet
Dress.
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation the most according to claim 1, it is special
Levy and be: the sterilizing in described step (2) is carried out in high-pressure sterilizing pot, sterilising conditions be 121 DEG C at a temperature of sterilizing 15 ~
20min。
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation the most according to claim 1, it is special
Levy and be: the composition of the beef extract-peptone fluid medium in described step (3) is: Carnis Bovis seu Bubali cream 5g, peptone 10g, chlorination
Sodium 5g, distilled water 1000mL, its pH is 7.2 ~ 7.5.
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation the most according to claim 1, it is special
Levy and be: the frequency of vibration of the constant temperature oscillator in described step (4) is 200 ~ 220r/min.
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation the most according to claim 1, it is special
To levy and be: in described step (6), the rotating speed of centrifuge used by centrifugal treating is 3500 ~ 4000r/min, centrifugation time is 10 ~
15min。
A kind of method utilizing alcohol industry yellow fluid to produce lichens bacillus preparation the most according to claim 1, it is special
Levy and be: the excipient described in described step (9) is lactose and soluble starch, and according to the weight ratio with described dry powder,
Its addition is lactose and the soluble starch of 20% of 4%.
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| CN101986865A (en) * | 2009-07-30 | 2011-03-23 | 天津瑞普生物技术股份有限公司 | Preparation of Bacillus licheniformis preparation for poultry |
| CN101812390A (en) * | 2010-04-02 | 2010-08-25 | 河南省宋河酒业股份有限公司 | Comprehensive utilization technology for waste discoloured water of brewing |
| CN102321516A (en) * | 2011-08-31 | 2012-01-18 | 重庆诗仙太白酒业（集团）有限公司 | Processing technology for secondary utilization of organic matter in solid-state fermentation of Luzhou-flavour liquor |
| CN103141666A (en) * | 2013-03-28 | 2013-06-12 | 山东轻工业学院 | Method for producing microbe feed probiotics by using white spirit vinasse |
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