CN104004677A - Method for production of bacillus licheniformis preparation by utilizing ethanol industrial yellow water - Google Patents
Method for production of bacillus licheniformis preparation by utilizing ethanol industrial yellow water Download PDFInfo
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Abstract
A method for production of a bacillus licheniformis preparation by utilizing ethanol industrial yellow water includes collection and treatment of the ethanol industrial yellow water, cultivation of bacillus licheniformis in the ethanol industrial yellow water, and receiving and preparation of bacillus licheniformis. The method for production of the bacillus licheniformis by utilizing the ethanol industrial yellow water not only can make full use of the ethanol industrial yellow water, reduce the treatment cost of the yellow water, optimize comprehensive utilization of the yellow water and improve the ecological environment, but also can expand sources of a nutrition substrate in a conventional bacillus licheniformis preparation production, lower the production costs and expand the application scope of bacillus licheniformis.
Description
Technical field
The comprehensive utilization field and the microorganism fermentation field that the present invention relates to alcohol industry yellow water, especially utilize the yellow water in alcohol industry to carry out microorganism fermentation.
Background technology
Alcohol industry is important energy industry of China, but can produce huge by product in alcohol production process, wherein the solids component such as vinasse can continue to produce the feedstuff raw materials such as DDGS and again be utilized together with part yellow water, and the yellow water of the overwhelming majority is not well utilized.Yellow water is the liquid substance that is rich in multiple nutritional components of discharging in producing alcohol process, in yellow water containing abundant organic acid (lactic acid especially, butyric acid, caproic acid, the carboxylic acid contents such as acetic acid are maximum), alcohol, aldehyde, esters etc. are fragrance matter, also contain amino acid simultaneously, carbohydrate, alcohol, soil ulmin, yeast thalline autolyzate, microbial cells and viable cell etc., because it is nutritious, directly enter the pollution that tends to cause serious body eutrophication in environment, for this reason, the waste water of national regulation alcohol enterprise discharge must be processed through early stage, and can discharge after reaching 4 grades of water quality standards, so, nutritive ingredient in yellow water is wasted, alcohol enterprise often increases more expense for the processing of yellow water, and the most of brewery of China is all to use it for mix grain time cellar for storing things fermentation or support cellar for storing things to the processing of yellow water at present, cultivate the aspects such as artificial distiller's yeast, not only can not efficent use of resources, and cannot fundamentally solve the final outlet problem of fermentation yellow water.Therefore how to fully utilize yellow water resource, how can turn waste into wealth, the outlet problem that thoroughly solves yellow water seems particularly important, if yellow water can be turned waste into wealth, fully utilize, the cost that reduces enterprise is dropped into, also meet the needs of improvement of the ecological environment simultaneously.
Bacillus licheniformis is Gram-positive bacillus, cell size 0.8 * (1.5 ~ 3.5) m.Bacillus licheniformis cellular form and arrangement are shaft-like, Dan Sheng, and its gemma form is for producing nearly middle raw ellipticity gemma, and sporangiocyst slightly expands.Bacillus licheniformis enters after enteron aisle as active bacteria formulation, the pathogenic bacterium such as staphylococcus, yeast-like fungus are had to antagonistic action, and the probioticss such as bifidus bacillus, lactobacillus, bacterioide, digestibility suis are had to promoter action, thereby reach, regulate microbial population of animal intestinal tract balance, improve the effect of enteric microorganism environment; Can stimulating animal immune organ grow, activated lymphocyte, improves immunoglobulin (Ig) and antibody horizontal, strengthens cellular immunization and humoral immune function, improves immunity of organisms; In addition, under optimum conditions, can synthetic proteins enzyme, the enzyme such as lipase, amylase and cellulase, jointly play a role with endogenous enzyme, promote digesting and assimilating of nutrient.Based on above feature, Bacillus licheniformis is widely applied in industries such as medicine, health care, food, biological fodder productions at present.Yet, the production method of Bacillus licheniformis preparation adopts people for adding a large amount of high-quality nutritive substances to produce as raw material more at present, its production cost is higher, and then has limited the application of Bacillus licheniformis, and increases and take the cost of the related products that Bacillus licheniformis is raw material.
Owing to being rich in the nutritive substance of multiple soluble state in alcohol industry yellow water, therefore, if can utilize alcohol industry yellow water to carry out the production of Bacillus licheniformis, can solve the problem of complex utilization of alcohol industry yellow water, turn waste into wealth, can reduce the production cost of current Bacillus licheniformis preparation again, further expand the range of application of Bacillus licheniformis.
Summary of the invention
The object of this invention is to provide a kind of production method of utilizing alcohol industry yellow water to carry out Bacillus licheniformis preparation, reduce on the one hand link and the expense that in alcohol industry, yellow water is processed, the comprehensive utilization ratio that improves alcohol industry yellow water, reduces the industrial cost of Bacillus licheniformis preparation on the other hand.
The technical scheme that the present invention solves the problems of the technologies described above employing is: a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation, and its concrete steps are as follows:
(1) from the sheet frame of plate-and-frame filter press, directly collect alcohol industry yellow water, put into rapidly the container of sealing, then the mensuration of the nutrient composition contents such as protein, Mierocrystalline cellulose, amino acid, organic acid, mineral element is carried out in sampling, and according to the nutritional need of practical measurement value and Bacillus licheniformis growth and breeding, carry out nutritive ingredient and supplement;
(2) by the alcohol industry yellow water after supplementing the nutrients in step (1) in high-pressure sterilizing pot under 121 ℃ of conditions sterilizing 15 ~ 20min, and carry out cooling, alcohol industry yellow water pH after sterilizing is 3.2 ~ 3.5, be not suitable for lichens genus bacillus growth and breeding, then utilize sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5;
(3) Bacillus licheniformis is inoculated in beef extract-peptone liquid nutrient medium, is cultured to the stably manufactured phase, detect viable count and be not less than 1 * 10
9cfu/mL can be used as seed bacterium liquid, standby, and wherein, the proportioning of described beef extract-peptone liquid nutrient medium is: extractum carnis 5g, peptone 10g, sodium-chlor 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5;
(4) by resulting kind of daughter bacteria liquid in step (3) according to 6% ~ 8%(v/v) ratio be inoculated in the alcohol industry yellow water after step (2) is processed, and at the temperature of 36 ℃ ~ 36.5 ℃, be placed on the constant temperature oscillator of 200 ~ 220r/min and cultivate 16 ~ 18h, obtain Bacillus licheniformis liquid, standby;
(5) in the Bacillus licheniformis liquid obtaining, add calcium chloride in step (4), the amount of its interpolation is 0.05% ~ 0.08% according to weightmeasurement ratio (w/v), then in 60 ℃ of water-baths, heat 10min ~ 20min, proceed to again in 80 ℃ of water-baths and heat 5min ~ 10min, obtain the bacterium liquid that contains brood cell, the Bacillus licheniformis spore forming rate 89.1% ~ 92.6% that the method is processed;
(6) by the bacterium liquid that contains gemma in step (5) with large capacity whizzer in the centrifugal 10 ~ 15min of 3500 ~ 4000r/min, abandoning supernatant, obtains Bacillus licheniformis solids;
(7) according to the Bacillus licheniformis solids of step (6) gained and the weight ratio of carrier 1:4 ~ 6, add Paris white carrier, mix, make Bacillus licheniformis solid mixture;
(8) the Bacillus licheniformis solid mixture of step (7) gained is dried under aseptic, 60 ℃ ~ 65 ℃ conditions, grind to form dry powder, in dry powder, effectively Bacillus licheniformis number is 20 * 10
9~ 30 * 10
9individual/g, effectively Bacillus licheniformis number refers to Bacillus licheniformis viable count and counts sum with the Bacillus licheniformis that forms gemma;
(9) in the resulting Bacillus licheniformis dry powder of step (8), according to weight ratio, add 4% lactose, 20% Zulkovsky starch as vehicle, mix, make Bacillus licheniformis preparation, effective Bacillus licheniformis number >=5,000,000,000/gram wherein, then with laminated film to lichens bacillus preparation, carry out vacuum packaging, vacuum tightness is 0.085 ~ 0.095Mpa.
The present invention Bacillus licheniformis used, purchased from Sang Ge bio tech ltd, Shanghai, is preserved in the biological product of USS collecting center, and its deposit number is ATCC 11946, and buying the date is on November 20th, 2013.
The present invention has following beneficial effect:
First, compare with nutraceutical matrix with the raw material that current Bacillus licheniformis preparation is produced, the present invention adopts alcohol industry yellow water as cultivating raw material and matrix, take full advantage of alcohol industry waste water, improved the comprehensive utilization value of alcohol industry yellow water, improve ecotope, expanded the source of Bacillus licheniformis preparation production desired nutritional matrix, reduced the production cost of Bacillus licheniformis;
Second, cultivation through Bacillus licheniformis, nutritive substance in alcohol industry yellow water is made full use of by Bacillus licheniformis, content effectively reduces, substantially reach the emission standard of alcohol waste water, can not cause the eutrophication of water body, and save link and relevant input of business processes alcohol industry yellow water, reduce the production cost of relevant enterprise;
The 3rd, the present invention directly gathers alcohol industry yellow water from the sheet frame of plate-and-frame filter press, this is because the bleeding point outbalance of alcohol industry yellow water, the yellow water nutritive ingredient that the yellow water nutritive ingredient of directly collecting from the sheet frame of plate-and-frame filter press is collected compared with water port is abundant, is more suitable as culture medium;
The 4th, the present invention is measuring under the prerequisite of alcohol industry yellow water nutrient contg, according to measurement result, targetedly yellow water is carried out to nutritional supplementation, to be more applicable to the Growth and reproduction of Bacillus licheniformis, further improved the output of Bacillus licheniformis in alcohol industry yellow water;
The 5th, for the distinctive chemical constitution of alcohol industry yellow water, carry out the change of suitable pre-treatment and part chemical property, make its Growth and reproduction that is more suitable for lichens genus bacillus, improve the output of Bacillus licheniformis;
The 6th, for peculiar chemical constitution and the character of alcohol industry yellow water, provide suitable to cultivating the operating parameters of Bacillus licheniformis, comprise Bacillus licheniformis inoculum size, culture temperature, shaking speed and incubation time etc.;
The 7th, after Bacillus licheniformis is cultivated and to finish, utilize provided by the invention collect and preparation method can make effective bacterium number >=5,000,000,000/gram of Bacillus licheniformis preparation, spore forming rate reaches 89.1% ~ 92.6%.
Therefore, the method of utilizing alcohol industry yellow water to produce Bacillus licheniformis provided by the present invention, can make full use of alcohol industry yellow water, reduce the processing cost of yellow water, optimize the comprehensive utilization of yellow water, improve the ecological environment, can expand again the source of nutraceutical matrix in existing Bacillus licheniformis preparation production, reduce production costs, expand the range of application of Bacillus licheniformis.
Accompanying drawing explanation
Fig. 1 is the content of various nutritive ingredients in alcohol industry yellow water.
Fig. 2 is the impacts of different culture temperature on effective Bacillus licheniformis number.
Fig. 3 is the impacts of different incubation times on effective Bacillus licheniformis number.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1: a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation, and what present method adopted is that corn is the yellow water of raw material production alcohol, concrete steps are as follows:
(1) from the sheet frame of plate-and-frame filter press, directly collect alcohol industry yellow water, put into rapidly the container of sealing, then protein is carried out in sampling, Mierocrystalline cellulose, amino acid, organic acid, the mensuration of the nutrient composition contents such as mineral element, obtain the content of various nutritive ingredients in alcohol industry yellow water, measurement result is shown in accompanying drawing 1, and according to the nutritional need of practical measurement value and Bacillus licheniformis growth and breeding, carrying out nutritive ingredient supplements, concrete nutritive ingredient supplements kind and quantity is: often go up in the alcohol industry yellow water of stating and add urea 5g, glucose 2g and sodium-chlor 2g,
(2) by the alcohol industry yellow water after supplementing the nutrients in step (1) in high-pressure sterilizing pot under 121 ℃ of conditions sterilizing 15 ~ 20min, and carry out coolingly, then utilize sodium hydroxide, sodium bicarbonate by its pH regulator to 7.2 ~ 7.5;
(3) Bacillus licheniformis is inoculated in beef extract-peptone liquid nutrient medium, is cultured to the stably manufactured phase, detect viable count and be not less than 1 * 10
9cfu/mL can be used as seed bacterium liquid, standby, and wherein, the proportioning of described beef extract-peptone liquid nutrient medium is: extractum carnis 5g, peptone 10g, sodium-chlor 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5;
(4) by resulting kind of daughter bacteria liquid in step (3) according to 6%(v/v) ratio be inoculated in the alcohol industry yellow water after step (2) is processed, and at the temperature of 36 ℃, be placed on the constant temperature oscillator of 200 ~ 220r/min and cultivate 16h, obtain Bacillus licheniformis liquid, standby;
(5) in the Bacillus licheniformis liquid obtaining, add calcium chloride in step (4), the amount of its interpolation is 0.05% according to weightmeasurement ratio (w/v), then in 60 ℃ of water-baths, heat 10min, then proceed in 80 ℃ of water-baths and heat 5min, obtain the bacterium liquid that contains brood cell;
(6) by the bacterium liquid that contains gemma in step (5) with large capacity whizzer in the centrifugal 10 ~ 15min of 3500 ~ 4000r/min, abandoning supernatant, obtains Bacillus licheniformis solids;
(7) according to the Bacillus licheniformis solids of step (6) gained and the weight ratio of carrier 1:4, add Paris white carrier, mix, make Bacillus licheniformis solid mixture;
(8) the Bacillus licheniformis solid mixture of step (7) gained is dried under aseptic, 60 ℃ ~ 65 ℃ conditions, grind to form dry powder;
(9) in the Bacillus licheniformis dry powder obtaining in step (8), according to weight ratio, add 4% lactose, 20% Zulkovsky starch as vehicle, mix, make Bacillus licheniformis preparation, then with laminated film, lichens bacillus preparation is carried out to vacuum packaging, vacuum tightness is 0.085 ~ 0.095Mpa.
After measured, adopting the effective bacterium number of Bacillus licheniformis in the method gained Bacillus licheniformis preparation in the present embodiment is (41.5 ± 5.3) * 10
9.
Embodiment 2: a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation, and what present method adopted is that corn is the yellow water of raw material production alcohol, concrete steps are as follows:
(1) from the sheet frame of plate-and-frame filter press, directly collect alcohol industry yellow water, put into rapidly the container of sealing, then protein is carried out in sampling, Mierocrystalline cellulose, amino acid, organic acid, the mensuration of the nutrient composition contents such as mineral element, obtain the content of various nutritive ingredients in alcohol industry yellow water, measurement result is shown in accompanying drawing 1, and according to the nutritional need of practical measurement value and Bacillus licheniformis growth and breeding, carrying out nutritive ingredient supplements, concrete nutritive ingredient supplements kind and quantity is: often go up in the alcohol industry yellow water of stating and add urea 5g, glucose 2g, sodium-chlor 2g,
(2) by the alcohol industry yellow water after supplementing the nutrients in step (1) in high-pressure sterilizing pot under 121 ℃ of conditions sterilizing 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 liquid nutrient medium, is cultured to the stably manufactured phase, detect viable count and be not less than 1 * 10
9cfu/mL can be used as seed bacterium liquid, standby, and wherein, the proportioning of described beef extract-peptone liquid nutrient medium is: extractum carnis 5g, peptone 10g, sodium-chlor 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5;
(4) by resulting kind of daughter bacteria liquid in step (3) according to 8%(v/v) ratio be inoculated in the alcohol industry yellow water after step (2) is processed, and at the temperature of 36.5 ℃, be placed on the constant temperature oscillator of 200 ~ 220r/min and cultivate 18h, obtain Bacillus licheniformis liquid, standby;
(5) in the Bacillus licheniformis liquid obtaining, add calcium chloride in step (4), the amount of its interpolation is 0.08% according to weightmeasurement ratio (w/v), then in 60 ℃ of water-baths, heat 20min, then proceed in 80 ℃ of water-baths and heat 10min, obtain the bacterium liquid that contains brood cell;
(6) by the bacterium liquid that contains gemma in step (5) with large capacity whizzer in the centrifugal 10 ~ 15min of 3500 ~ 4000r/min, abandoning supernatant, obtains Bacillus licheniformis solids;
(7) according to the Bacillus licheniformis solids of step (6) gained and the weight ratio of carrier 1:6, add Paris white, mix, make Bacillus licheniformis solid mixture;
(8) the Bacillus licheniformis solid mixture of step (7) gained is dried under aseptic, 60 ℃ ~ 65 ℃ conditions, grind to form dry powder;
(9) will in above-mentioned Bacillus licheniformis dry powder, according to weight ratio, add 4% lactose, 20% Zulkovsky starch as vehicle, mix, make Bacillus licheniformis preparation, then with laminated film, lichens bacillus preparation is carried out to vacuum packaging, vacuum tightness is 0.085 ~ 0.095Mpa.
After measured, adopting the effective bacterium number of Bacillus licheniformis in the method gained Bacillus licheniformis preparation in the present embodiment is (48.2 ± 4.3) * 10
9.
Embodiment 3: a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation, and what present method adopted is that corn is the yellow water of raw material production alcohol, concrete steps are as follows:
(1) from the sheet frame of plate-and-frame filter press, directly collect alcohol industry yellow water, put into rapidly the container of sealing, then protein is carried out in sampling, Mierocrystalline cellulose, amino acid, organic acid, the mensuration of the nutrient composition contents such as mineral element, obtain the content of various nutritive ingredients in alcohol industry yellow water, measurement result is shown in accompanying drawing 1, and according to the nutritional need of practical measurement value and Bacillus licheniformis growth and breeding, carrying out nutritive ingredient supplements, concrete nutritive ingredient supplements kind and quantity is: often go up in the alcohol industry yellow water of stating and add urea 5g, glucose 2g, sodium-chlor 2g,
(2) by the alcohol industry yellow water after supplementing the nutrients in step (1) in high-pressure sterilizing pot under 121 ℃ of conditions sterilizing 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 liquid nutrient medium, is cultured to the stably manufactured phase, detect viable count and be not less than 1 * 10
9cfu/mL can be used as seed bacterium liquid, standby, and wherein, the proportioning of described beef extract-peptone liquid nutrient medium is: extractum carnis 5g, peptone 10g, sodium-chlor 5g, distilled water 1000mL, and by its pH regulator to 7.2 ~ 7.5;
(4) by resulting kind of daughter bacteria liquid in step (3) according to 7%(v/v) ratio in the alcohol industry yellow water after step (2) is processed, and at the temperature of 36.2 ℃, be placed on the constant temperature oscillator of 200 ~ 220r/min and cultivate 17h, obtain Bacillus licheniformis liquid, standby;
(5) in the Bacillus licheniformis liquid obtaining, add calcium chloride in step (4), the amount of its interpolation is 0.07% according to weightmeasurement ratio (w/v), then in 60 ℃ of water-baths, heat 15min, then proceed in 80 ℃ of water-baths and heat 8min, obtain the bacterium liquid that contains brood cell;
(6) by the bacterium liquid that contains gemma in step (5) with large capacity whizzer in the centrifugal 10 ~ 15min of 3500 ~ 4000r/min, abandoning supernatant, obtains Bacillus licheniformis solids;
(7) according to the Bacillus licheniformis solids of step (6) gained and the weight ratio of carrier 1:5, add Paris white, mix, make Bacillus licheniformis solid mixture;
(8) the Bacillus licheniformis solid mixture of step (7) gained is dried under aseptic, 60 ℃ ~ 65 ℃ conditions, grind to form dry powder;
(9) will in above-mentioned Bacillus licheniformis dry powder, according to weight ratio, add 4% lactose, 20% Zulkovsky starch as vehicle, mix, make Bacillus licheniformis preparation, then with laminated film, lichens bacillus preparation is carried out to vacuum packaging, vacuum tightness is 0.085 ~ 0.095Mpa.
After measured, adopting the effective bacterium number of Bacillus licheniformis in the method gained Bacillus licheniformis preparation in the present embodiment is (45.6 ± 6.4) * 10
9.
Below in conjunction with test example, the present invention is further illustrated.
Test example 1: inquire into and adopt method provided by the present invention to carry out Bacillus licheniformis cultivation, the impact of different culture temperature on effective bacterium number in final Bacillus licheniformis finished product, and find out optimum culture temperature.
In the cultivation of microorganism, the impact of temperature is most important, directly has influence on the success or not of microorganism culturing, and whether final colony counts meets the requirements.According to method provided by the invention, adopt single factor design, culture temperature is set to respectively to the differing temps gradient of 35 ℃, 35.5 ℃, 36 ℃, 36.5 ℃, 37 ℃, 37.5 ℃, 39 ℃ and 40 ℃, carries out the cultivation of Bacillus licheniformis, test-results is shown in accompanying drawing 2.
In accompanying drawing 2, can find out, while carrying out the cultivation of Bacillus licheniformis according to method of the present invention, when culture temperature is during at 36 ℃ to 36.5 ℃, in Bacillus licheniformis preparation, effectively bacterium number is higher, especially 36.5 ℃ time, effectively bacterium number is the highest, and therefore, 36.5 ℃ is to adopt method of the present invention to cultivate the optimum culture temperature of Bacillus licheniformis.
Test example 2: inquire into and adopt method provided by the present invention to carry out Bacillus licheniformis cultivation, the impact of different incubation times on effective bacterium number in final lichens bacillus preparation, and find out optimum incubation time.
In microorganism culturing, the too short bacterium colony that may can not get effective quantity of time, overlong time may be by living contaminants, and therefore, incubation time is most important.According to method of the present invention, adopt single factor design, incubation time is set to respectively to 12h, 16h, 18h, 24h, 36h and 48h, carry out the cultivation of Bacillus licheniformis, test-results is shown in accompanying drawing 3.
From accompanying drawing 3, can find out, when incubation time is 16 ~ 18h, in Bacillus licheniformis preparation, effectively bacterium quantity is more, especially when incubation time is 18h, the effective bacterium number of Bacillus licheniformis reaches the highest, and therefore according to method of the present invention, optimum incubation time is 18h.
Claims (6)
1. utilize alcohol industry yellow water to produce a method for lichens bacillus preparation, it is characterized in that: comprise the following steps:
(1) the alcohol industry yellow water of collecting is put into the container of sealing, the mensuration of nutritive ingredient is carried out in sampling, and according to measured value, collected alcohol industry yellow water is carried out to nutritive ingredient and supplement, standby;
(2) will in step (1), through the supplementary alcohol industry yellow water of nutritive ingredient, carry out sterilizing, pH regulator to 7.2 ~ 7.5, standby;
(3) Bacillus licheniformis is cultured in beef extract-peptone liquid nutrient medium to the stably manufactured phase, detects viable count and be not less than 1 * 10
9cfu/mL is as kind of a daughter bacteria liquid, standby;
(4) by resulting kind of daughter bacteria liquid in step (3) according to 6% ~ 8%(v/v) ratio be inoculated in through step (2) and process in the alcohol industry yellow water of gained, and under the culture temperature of 36 ℃ ~ 36.5 ℃, on constant temperature oscillator, cultivate 16h ~ 18h, obtain Bacillus licheniformis liquid, standby;
(5) in the Bacillus licheniformis liquid obtaining, add calcium chloride in step (4), the amount of its interpolation is 0.05% ~ 0.08% according to weightmeasurement ratio (w/v), then in 60 ℃ of water-baths, heat 15min ~ 20min, proceed to again in 80 ℃ of water-baths and heat 5min ~ 10min, obtain the bacterium liquid that contains brood cell;
(6) the bacterium liquid that contains gemma in step (5) is carried out to centrifugal treating with whizzer, abandoning supernatant, obtains Bacillus licheniformis solids;
(7) according to the Bacillus licheniformis solids of step (6) gained and the part by weight of carrier 1:4 ~ 6, add Paris white carrier, and mix, make Bacillus licheniformis solid mixture;
(8) by the Bacillus licheniformis solid mixture obtaining in step (7) under aseptic condition, in 60 ℃ ~ 65 ℃ oven dry, grind to form dry powder;
(9) in the dry powder obtaining in step (8), add vehicle, mix, make Bacillus licheniformis preparation, and vacuum packaging.
2. a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation according to claim 1, is characterized in that: the sterilizing in described step (2) is carried out in high-pressure sterilizing pot, and sterilising conditions is sterilizing 15 ~ 20min at 121 ℃ of temperature.
3. a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation according to claim 1, it is characterized in that: the formula of the beef extract-peptone liquid nutrient medium in described step (3) is: extractum carnis 5g, peptone 10g, sodium-chlor 5g, distilled water 1000mL, its pH is 7.2 ~ 7.5.
4. a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation according to claim 1, is characterized in that: the vibrational frequency of the constant temperature oscillator in described step (4) is 200 ~ 220r/min.
5. a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation according to claim 1, is characterized in that: the rotating speed of the whizzer that the middle centrifugal treating of described step (6) is used is 3500 ~ 4000r/min, and centrifugation time is 10 ~ 15min.
6. a kind of method of utilizing alcohol industry yellow water to produce lichens bacillus preparation according to claim 1, it is characterized in that: the vehicle described in described step (9) is lactose and Zulkovsky starch, and according to the weight ratio with described dry powder, the lactose that its addition is 4% and 20% Zulkovsky starch.
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| CN115094006A (en) * | 2022-07-08 | 2022-09-23 | 贵州茅台酒股份有限公司 | Resource utilization method of vinasse leachate |
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| CN104522053B (en) * | 2014-12-31 | 2017-10-13 | 金云初 | A kind of disinfectant for vegetables of high-molecular biologic sustained release and preparation method thereof |
| CN115094006A (en) * | 2022-07-08 | 2022-09-23 | 贵州茅台酒股份有限公司 | Resource utilization method of vinasse leachate |
| CN115094006B (en) * | 2022-07-08 | 2024-04-16 | 贵州茅台酒股份有限公司 | Resource utilization method of vinasse percolate |
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