CN106906168B - A kind of spirit stillage decomposing agent and its preparation method and application - Google Patents
A kind of spirit stillage decomposing agent and its preparation method and application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C05—FERTILISERS; MANUFACTURE THEREOF
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- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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Abstract
The invention discloses a kind of spirit stillage decomposing agents, thermoactinomyces sacchari CP with high temperature resistant, ethanol-tolerant, acid and alkali-resistance characteristic is learned with existing decomposed Cordycepps and compound, using the synergistic effect between different decomposed bacterium simultaneously to ingredients progress fully degradeds such as cellulose, lignin, starch, protein, fat in vinasse.Without carrying out the drop alcoholic strength of vinasse, the processing such as deacidification degree can direct compost fermentation fresh spirit stillage, reduce treatment process, manpower is saved, material resources, financial resources and time, into the megathermal period, short (1d reaches 55 DEG C, 3d reaches 85 DEG C), maintain the hot fermentation time long (83-85 DEG C of maintenance 3d), shorten fermentation period (only 15-16d), shorten 26-27 days than existing spirit stillage decomposing agent, effectively increase the yield and quality (rotten degree) of organic fertilizer, prevent the secondary pollution corruption and alkalization of soils of vinasse, protect environment and soil, a new shortcut is opened for the decomposed production high quality organic fertilizer of the direct During High-Temperature Composting of spirit stillage.
Description
Technical field
The present invention relates to vinasse decomposing agent, in particular to a kind of spirit stillage decomposing agent and its preparation method and application.
Background technique
Vinasse are the direct leftover bits and pieces during wine brewing, and China's vinasse to be treated are up to 50,000,000 tons or more at present, because
This, has always been concerned with the vinasse processing problem after fermentation.Spirit stillage, easily fermentation corruption, it is easy to distribute stench
It pollutes the environment.Because it contains a large amount of crude fat, Crude starch, crude protein and the ingredients such as N P and K abundant and polysaccharide,
It is fabulous organic fertilizer resource.Therefore organic fertilizer is made using vinasse, both can solve environmental issue, turn waste into wealth, and can is
Green agriculture production provides high-quality organic fertilizer, reduces the usage amount of chemical fertilizer, has high economic benefit, environmental benefit and society
It can benefit.
It is the most important mode of current vinasse production organic fertilizer that vinasse During High-Temperature Composting is decomposed, and vinasse decomposing agent is vinasse
The critical materials of organic fertilizer manufacturing process are the important leverages of final product quality and benefit, and still, spirit stillage is different from other
The important feature of organic fertilizer fermentation matrix is: fresh vinasse water content big (60-65%), acidity height (pH value 3.4-4.0), wine
Precision height (6-10%) is unfavorable for the growth and breeding of general decomposed bacterium, it is decomposed to be difficult direct fermentation.Currently, related white wine
The patent document of vinasse decomposing agent is more:
106146105 A of Chinese patent CN discloses a kind of cassava grain stillage organic fertilizer containing amino acid, which includes
The raw material of following mass fraction: 800-900 parts of cassava grain stillage, 175-250 parts of plant ash, 50-100 parts of vegetable protein, compound bacteria
0.2-0.4 parts of kind；The composite bacteria is made of heat-resistant bacillus and Mei Jiulan streptomycete by the mass ratio of 1:1-1.2.
104151042 B of Chinese patent CN discloses a kind of vinasse decomposing agent preparation method, the master of the decomposing microbial inoculum
It is configured to the bacillus amyloliquefaciens that deposit number is CGMCC No.8143 and the not sea that deposit number is CGMCCNo.8268
Prestige bacillus；Its compound proportion is 3:7.Effective bacterium of decomposing agent microbial inoculum living >=2 × 108cfu/g。
106278526 A of Chinese patent CN discloses a kind of preparation method of microorganism-decomposing agent, and the high temperature is decomposed
Agent is liquid microbial inoculum, and the viable bacteria content of the liquid microbial inoculum is hundred million/ml of 1.8-2.5；The strain of the liquid microbial inoculum includes withered grass
Bacillus, actinomyces and saccharomycete, the bacillus subtilis, actinomyces, saccharomycete strain quantity ratio be 3:4:3, institute
The weight for stating the addition of high temperature decomposing agent is the 0.4-0.6% for the total weight that fertilizer matrix adjusts after moisture content and pH value.
104909854 A of Chinese patent CN discloses a kind of sauce incense liquor grain decomposition agent strain composition, the decomposing microbial inoculum
It is combined by bacillus, saccharomycete, actinomyces and acetobacter, but there is no specific open compound proportion, decomposing agents
Effective bacterium of microbial inoculum living >=0.5 × 108cfu/g。
Spirit stillage high temperature decomposing microbial inoculum disclosed above is although more, but requires to carry out the drop alcoholic strength of vinasse, drop
Acidity, addition aid nutrition are pre-processed at grading mode, complex procedures, waste of manpower, material resources, financial resources and time, and can also
Causing the decomposition of vinasse to be not thorough causes secondary pollution corrupt, and the additive amount for existing simultaneously decomposing agent is higher, and fermentation rate is low, high
The defects of warm compost maturity period is long, and organic matter degradation rate is low, decomposed incomplete, therefore it provides a kind of pre- without carrying out vinasse
Processing and composition adjustment, can directly be inoculated with spirit stillage and carry out the decomposed quick composting microbial inoculum of During High-Temperature Composting is art technology
Personnel there is an urgent need to.
Summary of the invention
Technical problem solved by the invention is the shortcomings that overcoming existing spirit stillage decomposing agent, according to spirit stillage at
It is grouped as and nutritive peculiarity, it will be with high temperature resistant, ethanol-tolerant, the thermoactinomyces sacchari CP of acid and alkali-resistance characteristic and existing decomposed Cordycepps
Compound, and provides one kind and is not necessarily to carry out vinasse deacidification, pre-process alcohol etc., quick, the direct decomposed fresh spirit stillage of energy
Decomposing agent.
In order to achieve the above object, the invention adopts the following technical scheme:
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
1-10 parts of thermoactinomyces sacchari microbial inoculum, 1-5 parts of bacillus licheniformis agent, 1-5 parts of bacillus subtilis microbial agent, solution
1-5 parts of bacillus amyloliquefaciens microbial inoculum；
Preferably, the spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
3-7 parts of thermoactinomyces sacchari microbial inoculum, 2-4 parts of bacillus licheniformis agent, 2-4 parts of bacillus subtilis microbial agent, solution
2-4 parts of bacillus amyloliquefaciens microbial inoculum；
It is highly preferred that the spirit stillage decomposing agent, is mainly prepared by the raw material of following parts by weight:
5 parts of thermoactinomyces sacchari microbial inoculum, 3 parts of bacillus licheniformis agent, 3 parts of bacillus subtilis microbial agent, solve starch bud
3 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, is from Tang, Hebei province
It separates, purify in the decomposed object of mountain city cow dung high temperature, screening obtains.It is micro- that the bacterial strain has been preserved in China on March 24th, 2017
Biological inoculum preservation administration committee common micro-organisms center (abbreviation CGMCC, address are as follows: BeiChen West Road, Chaoyang District, BeiJing City 1
No. 3 Institute of Microorganism, Academia Sinica, institute, postcode: 100101), deposit number is CGMCC NO.13928, classification life
Entitled thermoactinomyces sacchari Laceyella sacchari；
Thermoactinomyces sacchari (Laceyella sacchari) CP can be grown within the scope of 30-75 DEG C, the most suitable growth
Temperature is 60 DEG C；It can produce a variety of degrading enzymes such as protease, lipase；
Thermoactinomyces sacchari (Laceyella sacchari) CP is in the various concentration that alcoholic strength (V/V) is 1-13%
NA fluid nutrient medium in 50 DEG C of culture 3d in NA plate and NA fluid nutrient medium test tube are coated with after 50 DEG C of culture 3d, can grow
Well, two tests prove that the bacterial strain can tolerate alcoholic strength range in 1-13%, have stronger alcoholic strength tolerance level；
NA liquid of thermoactinomyces sacchari (Laceyella sacchari) CP in the different pH values that pH value is 3-12
50 DEG C of culture 3d in NA plate and NA fluid nutrient medium test tube, energy well-grown are coated in body culture medium after 50 DEG C of culture 3d,
The range that two tests prove that the bacterial strain can tolerate pH value is 3-12, more wide in range, has stronger soda acid tolerance level.
Preferably, viable bacteria content >=5 × 10 of the thermoactinomyces sacchari microbial inoculum11cfu/g；
It is highly preferred that the preparation method of the thermoactinomyces sacchari microbial inoculum, includes the following steps: thermoactinomyces sacchari CP
Slant strains are inoculated in NA seed bottle solid medium with transfer needle, 58-62 DEG C of culture 22-26h, and by the sterile washing of thallus
De- to scrape, adjustment cell concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 0.8-1.2kg solid medium A,
58-62 DEG C of culture 45-50h, dry 22-26h, is crushed with pulverizer under the conditions of 55-65 DEG C, crosses 40 meshes up to sugarcane
The uncommon Salmonella CP microbial inoculum of orchid, wherein thermoactinomyces sacchari viable bacteria content is 5-7 × 1011cfu/g；
The NA seed bottle solid medium the preparation method comprises the following steps: take peptone 10g, powdered beef 3g, sodium chloride 10g, fine jade
Rouge 20g, distilled water 1L are uniformly mixed, and are sufficiently dissolved, and adjustment pH value is 7.0, are dispensed into seed bottle after boiling, every bottle of 80-
100mL, 121 DEG C of sterilizing 30min, is put into inclined-plane to obtain the final product；
The solid medium A's the preparation method comprises the following steps: take rice husk 100g, wheat bran 300g, corn flour 200g, beancake powder
100g, starch 100g, laterite 100g, calcium sulfate 5g, calcium oxide 7.5g, dipotassium hydrogen phosphate 3g, magnesium sulfate 2g, water 400mL, uniformly
Mixing, 121 DEG C of sterilizing 45min to obtain the final product.
Further, the bacillus licheniformis is bacillus licheniformis ATCC 27811, bacillus licheniformis CICC
10037, bacillus licheniformis CICC 10181, bacillus licheniformis CICC 10831, in bacillus licheniformis CICC 10291
Any one or a few；
Preferably, the bacillus licheniformis is bacillus licheniformis ATCC 27811；
Preferably, viable bacteria content >=3 × 10 of the bacillus licheniformis agent11cfu/g；
It is highly preferred that the preparation method of the bacillus licheniformis agent, includes the following steps: bacillus licheniformis is oblique
Face strain is inoculated into NA seed bottle culture medium with transfer needle, 1d is cultivated at 50 DEG C, and thallus is scraped with sterile water elution, adjusts
Whole cell concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium B, 50 DEG C of culture 2d, 50
It is crushed after dry 2d with pulverizer under the conditions of DEG C, crosses 40 meshes up to bacillus licheniformis agent, wherein lichens gemma bar
Bacterium viable bacteria content is 5 × 1011cfu/g；
Solid medium B's the preparation method comprises the following steps: take wheat bran 800g, powdered rice hulls 100g, beancake powder 5g, ammonium sulfate 5g,
Magnesium sulfate 1g, manganese sulfate 0.5g, water 1200mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
Further, the bacillus subtilis is bacillus subtilis ATCC 6051, bacillus subtilis CICC
10088, bacillus subtilis CICC 10090, bacillus subtilis CICC 10153, in bacillus subtilis CICC 10157
Any one or a few；
Preferably, the bacillus subtilis is bacillus subtilis ATCC 6051；
Preferably, bacillus subtilis microbial agent viable bacteria content >=4 × 1010cfu/g；
It is highly preferred that the preparation method of the bacillus subtilis microbial agent, includes the following steps: bacillus subtilis is oblique
Face strain is inoculated into NA seed bottle culture medium with transfer needle, 1d is cultivated at 37 DEG C, and thallus is scraped with sterile water elution, adjusts
Whole cell concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium C, 37 DEG C of culture 2d, 37
It is crushed after dry 3d with pulverizer under the conditions of DEG C, crosses 40 meshes up to bacillus subtilis microbial agent, wherein bacillus subtilis
Bacterium viable bacteria content is 6 × 1010cfu/g；
Solid medium C's the preparation method comprises the following steps: take wheat bran 800g, rice husk 100g, corn flour 50g, dregs of beans 50g, sulphur
Sour magnesium 0.5g, ammonium sulfate 5g, water 1100mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
Further, the bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10035, bacillus amyloliquefaciens
CICC 10036, bacillus amyloliquefaciens CICC 10074, bacillus amyloliquefaciens CICC 10163, bacillus amyloliquefaciens
Any one or a few in CICC 20027；
Preferably, the bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10035；
Preferably, viable bacteria content >=3 × 10 of the bacillus amyloliquefaciens microbial inoculum11cfu/g；
It is highly preferred that the preparation method of the bacillus amyloliquefaciens microbial inoculum, includes the following steps: that starch gemma bar will be solved
Bacterium slant strains are inoculated into NA seed bottle culture medium with transfer needle, 1d are cultivated at 37 DEG C, and thallus is scraped with sterile water elution
Under, adjustment cell concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium D, 37 DEG C of culture 2d,
It is crushed after 37 DEG C of dry 3d with pulverizer, crosses 40 meshes up to bacillus amyloliquefaciens microbial inoculum, wherein solution starch gemma bar
Bacterium viable bacteria content 5 × 1011cfu/g；
The solid medium D's the preparation method comprises the following steps: take groundnut meal 500g, wheat bran 350g, cotton dregs 120g, peptone
3g, glucose 2g, magnesium sulfate 0.2g, water 450mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
Another object of the present invention is to provide the preparation method of above-mentioned spirit stillage decomposing agent, includes the following steps: according to matching
Side, precise thermoactinomyces sacchari microbial inoculum, bacillus licheniformis agent, bacillus subtilis microbial agent, bacillus amyloliquefaciens
Microbial inoculum is uniformly mixed according to equal increments method up to spirit stillage decomposing agent.
The present invention also provides application of the above-mentioned spirit stillage decomposing agent in terms of quick composting spirit stillage, will be of the invention
The spirit stillage decomposing agent of preparation is decomposed to carry out in 1~1.5 ‰ fresh spirit stillage heap body of inoculum concentration access in mass ratio
Processing, 16d can complete compost maturity, reach innoxious standard, obtain up-to-standard lees organic fertilizer.
The utility model has the advantages that
Spirit stillage decomposing agent prepared by the present invention will have resistance to height according to the constituent structure and nutritive peculiarity of spirit stillage
Temperature, ethanol-tolerant, acid and alkali-resistance characteristic thermoactinomyces sacchari CP learn and compound with existing decomposed Cordycepps, utilize the association between the decomposed bacterium of difference
Same-action can carry out fully degraded to ingredients such as cellulose, lignin, starch, protein, fat in vinasse simultaneously, can have
Effect improves decomposing agent to the degradation capability of vinasse.Especially thermoactinomyces sacchari CP excellent ethanol-tolerant, acid and alkali-resistance characteristic, are not necessarily to
Carry out the processing such as the drop alcoholic strengths of vinasse, deacidification degree can the direct fresh spirit stillage of compost fermentation, reduce treatment process, save
It is about human and material resources, financial resources and time, into the megathermal period short (1d reaches 55 DEG C, and 3d reaches 85 DEG C), maintain the hot fermentation time
Long (83-85 DEG C of maintenance 3d), shortens fermentation period (only 15-16d), shortens 26-27 days than existing spirit stillage decomposing agent,
Effectively increase the yield and quality (rotten degree) of organic fertilizer, it is therefore prevented that the secondary pollution corruption and alkalization of soils of vinasse are protected
Environment and soil have been protected, has opened a new victory for the decomposed production high quality organic fertilizer of the direct During High-Temperature Composting of spirit stillage
Diameter.
Detailed description of the invention
Fig. 1 is the variation of temperature under different decomposed processing in spirit stillage composting process；
Fig. 2 is the variation of pH value under different decomposed processing in spirit stillage composting process；
Fig. 3 is the variation of the content of organic matter under different decomposed processing in spirit stillage composting process；
Fig. 4 is the variation of total nitrogen content under different decomposed processing in spirit stillage composting process；
Fig. 5 is the variation of moisture content under different decomposed processing in spirit stillage composting process.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.
Embodiment 1
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
5 parts of thermoactinomyces sacchari microbial inoculum, 3 parts of bacillus licheniformis agent, 3 parts of bacillus subtilis microbial agent, solve starch bud
3 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method of the thermoactinomyces sacchari microbial inoculum includes the following steps: thermoactinomyces sacchari CP slant strains
It is inoculated in NA seed bottle solid medium with transfer needle, 60 DEG C of cultures for 24 hours, and thallus are scraped with sterile water elution, adjust bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium A, 60 DEG C of culture 48h, in 60 DEG C of items
It is dry under part to be crushed for 24 hours with pulverizer, 40 meshes are crossed up to thermoactinomyces sacchari microbial inoculum, and wherein thermoactinomyces sacchari is living
Bacterial content is 7 × 1011cfu/g；
The NA seed bottle solid medium the preparation method comprises the following steps: take peptone 10g, powdered beef 3g, sodium chloride 10g, fine jade
Rouge 20g, distilled water 1L are uniformly mixed, and are sufficiently dissolved, and adjustment pH value is 7.0, are dispensed into seed bottle after boiling, every bottle of 80-
100mL, 121 DEG C of sterilizing 30min, is put into inclined-plane to obtain the final product；
The solid medium A's the preparation method comprises the following steps: take rice husk 100g, wheat bran 300g, corn flour 200g, beancake powder
100g, starch 100g, laterite 100g, calcium sulfate 5g, calcium oxide 7.5g, dipotassium hydrogen phosphate 3g, magnesium sulfate 2g, water 400mL, uniformly
Mixing, 121 DEG C of sterilizing 45min to obtain the final product.
The bacillus licheniformis is bacillus licheniformis ATCC 27811；
The preparation method of the bacillus licheniformis agent includes the following steps: to use bacillus licheniformis slant strains
Transfer needle is inoculated into NA seed bottle culture medium, 1d is cultivated at 50 DEG C, and thallus is scraped with sterile water elution, and adjustment thallus is dense
Degree is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium B, 50 DEG C of culture 2d, under the conditions of 50 DEG C
It is crushed after dry 2d with pulverizer, crosses 40 meshes up to bacillus licheniformis agent, wherein bacillus licheniformis viable bacteria contains
Amount is 5 × 1011cfu/g；
Solid medium B's the preparation method comprises the following steps: take wheat bran 800g, powdered rice hulls 100g, beancake powder 5g, ammonium sulfate 5g,
Magnesium sulfate 1g, manganese sulfate 0.5g, water 1200mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
The bacillus subtilis is bacillus subtilis ATCC 6051；
The preparation method of the bacillus subtilis microbial agent includes the following steps: to use bacillus subtilis slant strains
Transfer needle is inoculated into NA seed bottle culture medium, 1d is cultivated at 37 DEG C, and thallus is scraped with sterile water elution, and adjustment thallus is dense
Degree is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium C, 37 DEG C of culture 2d, under the conditions of 37 DEG C
It is crushed after dry 3d with pulverizer, crosses 40 meshes up to bacillus subtilis microbial agent, wherein bacillus subtilis viable bacteria contains
Amount is 6 × 1010cfu/g；
Solid medium C's the preparation method comprises the following steps: take wheat bran 800g, rice husk 100g, corn flour 50g, dregs of beans 50g, sulphur
Sour magnesium 0.5g, ammonium sulfate 5g, water 1100mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
The bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10035；
The preparation method of the bacillus amyloliquefaciens microbial inoculum includes the following steps: bacillus amyloliquefaciens inclined-plane bacterium
Kind is inoculated into NA seed bottle culture medium with transfer needle, 1d is cultivated at 37 DEG C, and thallus is scraped with sterile water elution, adjusts bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1kg solid medium D, 37 DEG C of culture 2d, 37 DEG C of dryings
It is crushed after 3d with pulverizer, crosses 40 meshes up to bacillus amyloliquefaciens microbial inoculum, wherein bacillus amyloliquefaciens viable bacteria contains
Amount 5 × 1011cfu/g；
The solid medium D's the preparation method comprises the following steps: take groundnut meal 500g, wheat bran 350g, cotton dregs 120g, peptone
3g, glucose 2g, magnesium sulfate 0.2g, water 450mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
Preparation method: according to formula, precise thermoactinomyces sacchari microbial inoculum, bacillus licheniformis agent, withered grass gemma
Bacillus microbial inoculum, bacillus amyloliquefaciens microbial inoculum are uniformly mixed according to equal increments method up to spirit stillage decomposing agent.
Embodiment 2
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
3 parts of thermoactinomyces sacchari microbial inoculum, 2 parts of bacillus licheniformis agent, 2 parts of bacillus subtilis microbial agent, solve starch bud
2 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method of the thermoactinomyces sacchari microbial inoculum includes the following steps: thermoactinomyces sacchari CP slant strains
NA seed bottle solid medium, 58 DEG C of culture 22h are inoculated in transfer needle, and thallus is scraped with sterile water elution, adjust bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 0.8kg solid medium A, 58 DEG C of culture 45h, at 55 DEG C
Under the conditions of dry 22h, crushed with pulverizer, cross 40 meshes up to thermoactinomyces sacchari microbial inoculum, wherein thermoactinomyces sacchari
Viable bacteria content is 6 × 1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC 10037；
The bacillus subtilis is bacillus subtilis CICC 10088；
The bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10036；
The bacillus licheniformis agent, bacillus subtilis microbial agent, bacillus amyloliquefaciens microbial inoculum preparation method and
Viable bacteria content is the same as embodiment 1；
In preparation process, NA seed bottle solid medium, solid medium B, solid medium C, is consolidated solid medium A
The preparation method is the same as that of Example 1 by body culture medium D；
The preparation method is the same as that of Example 1 for the spirit stillage decomposing agent.
Embodiment 3
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
7 parts of thermoactinomyces sacchari microbial inoculum, 4 parts of bacillus licheniformis agent, 4 parts of bacillus subtilis microbial agent, solve starch bud
4 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method of the thermoactinomyces sacchari microbial inoculum includes the following steps: thermoactinomyces sacchari CP slant strains
NA seed bottle solid medium, 62 DEG C of culture 26h are inoculated in transfer needle, and thallus is scraped with sterile water elution, adjust bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1.2kg solid medium A, 62 DEG C of culture 50h, at 65 DEG C
Under the conditions of dry 26h, crushed with pulverizer, cross 40 meshes up to thermoactinomyces sacchari microbial inoculum, wherein thermoactinomyces sacchari
Viable bacteria content is 5 × 1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC 10181；
The bacillus subtilis is bacillus subtilis CICC 10090；
The bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10074；
The bacillus licheniformis agent, bacillus subtilis microbial agent, bacillus amyloliquefaciens microbial inoculum preparation method and
Viable bacteria content is the same as embodiment 1；
In preparation process, NA seed bottle solid medium, solid medium B, solid medium C, is consolidated solid medium A
The preparation method is the same as that of Example 1 by body culture medium D；
The preparation method is the same as that of Example 1 for the spirit stillage decomposing agent.
Embodiment 4
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
1 part of thermoactinomyces sacchari microbial inoculum, 1 part of bacillus licheniformis agent, 1 part of bacillus subtilis microbial agent, solve starch bud
1 part of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method of the thermoactinomyces sacchari microbial inoculum includes the following steps: thermoactinomyces sacchari CP slant strains
NA seed bottle solid medium, 58 DEG C of culture 26h are inoculated in transfer needle, and thallus is scraped with sterile water elution, adjust bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 0.8kg solid medium A, 62 DEG C of culture 45h, at 65 DEG C
Under the conditions of dry 22h, crushed with pulverizer, cross 40 meshes up to thermoactinomyces sacchari microbial inoculum, wherein thermoactinomyces sacchari
Viable bacteria content is 6.2 × 1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC 10831；
The bacillus subtilis is bacillus subtilis CICC 10153；
The bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10163；
The bacillus licheniformis agent, bacillus subtilis microbial agent, bacillus amyloliquefaciens microbial inoculum preparation method and
Viable bacteria content is the same as embodiment 1；
In preparation process, NA seed bottle solid medium, solid medium B, solid medium C, is consolidated solid medium A
The preparation method is the same as that of Example 1 by body culture medium D；
The preparation method is the same as that of Example 1 for the spirit stillage decomposing agent.
Embodiment 5
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
10 parts of thermoactinomyces sacchari microbial inoculum, 5 parts of bacillus licheniformis agent, 5 parts of bacillus subtilis microbial agent, solve starch bud
5 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method of the thermoactinomyces sacchari microbial inoculum includes the following steps: thermoactinomyces sacchari CP slant strains
NA seed bottle solid medium, 62 DEG C of culture 22h are inoculated in transfer needle, and thallus is scraped with sterile water elution, adjust bacterium
Bulk concentration is 5 × 1011Cfu/mL draws 100mL bacterium solution and is inoculated into 1.2kg solid medium A, 58 DEG C of culture 50h, at 55 DEG C
Under the conditions of dry 26h, crushed with pulverizer, cross 40 meshes up to thermoactinomyces sacchari microbial inoculum, wherein thermoactinomyces sacchari
Viable bacteria content is 5.6 × 1011cfu/g；
The bacillus licheniformis is bacillus licheniformis ATCC 27811 and bacillus licheniformis CICC 10291；
The bacillus subtilis is bacillus subtilis ATCC 6051, bacillus subtilis CICC 10153 and withered grass
Bacillus CICC 10157；
The bacillus amyloliquefaciens are bacillus amyloliquefaciens CICC 10036, bacillus amyloliquefaciens CICC
10074, bacillus amyloliquefaciens CICC 10163 and bacillus amyloliquefaciens CICC 20027；
The bacillus licheniformis agent, bacillus subtilis microbial agent, bacillus amyloliquefaciens microbial inoculum preparation method and
Viable bacteria content is the same as embodiment 1；
In preparation process, NA seed bottle solid medium, solid medium B, solid medium C, is consolidated solid medium A
The preparation method is the same as that of Example 1 by body culture medium D；
The bacillus licheniformis agent is by 27811 microbial inoculum of bacillus licheniformis ATCC and bacillus licheniformis CICC
10291 microbial inoculums 2:1 in mass ratio is uniformly mixed；
The bacillus subtilis microbial agent is by 6051 microbial inoculum of bacillus subtilis ATCC, bacillus subtilis CICC
10153 microbial inoculums and 10157 microbial inoculum of bacillus subtilis CICC 1:1:1 in mass ratio are uniformly mixed；
The bacillus amyloliquefaciens microbial inoculum is by 10036 microbial inoculum of bacillus amyloliquefaciens CICC, bacillus amyloliquefaciens
10074 microbial inoculum of CICC, 10163 microbial inoculum of bacillus amyloliquefaciens CICC and 20027 microbial inoculum of bacillus amyloliquefaciens CICC press matter
Measure mixing more uniform than 4:3:2:1；
The preparation method is the same as that of Example 1 for the spirit stillage decomposing agent.
Embodiment 6
A kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight:
5 parts of thermoactinomyces sacchari microbial inoculum, 3 parts of bacillus licheniformis agent, 3 parts of bacillus subtilis microbial agent, solve starch bud
3 parts of spore bacillus microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, and deposit number is
CGMCC NO.13928, classification naming are thermoactinomyces sacchari Laceyella sacchari；
The preparation method and viable bacteria content of the thermoactinomyces sacchari microbial inoculum are the same as embodiment 1；
In preparation process, NA seed bottle solid medium, solid medium B, solid medium C, is consolidated solid medium A
The preparation method is the same as that of Example 1 by body culture medium D；
The bacillus licheniformis viable bacteria content is 3 × 1011cfu/g；
The bacillus subtilis viable bacteria content is 4 × 1010cfu/g；
The bacillus amyloliquefaciens viable bacteria content 3 × 1011cfu/g；
The preparation method is the same as that of Example 1 for the spirit stillage decomposing agent.
7 thermoactinomyces sacchari of embodiment (Laceyella sacchari) CP pH value tolerance test
The appropriate thallus of thermoactinomyces sacchari CP slant strains picking is accessed into 100mL NA fluid nutrient medium, 50 DEG C, 200r/
Min shaking table culture for 24 hours seed liquor, first with 1 ‰ inoculum concentrations access be tuned into respectively pH value be 1.5,2,2.5,3,3.5,4,
In 4.5,5,5.5,6,6.5,7,7.5,8,8.5,9,9.5,10,10.5,11,11.5,12 NA fluid nutrient medium, 50 DEG C,
It is coated with NA plate after 200r/min shaking table culture culture 3d, observes growing state；Then seed liquor is accessed with 1 ‰ inoculum concentrations again
The NA fluid nutrient medium test tube of same pH gradient, 50 DEG C, 200r/min shaking table culture 3d observe turbidity.
Test result: thermoactinomyces sacchari CP 50 DEG C of culture 3d in the NA fluid nutrient medium of pH value 3-12, coating NA are flat
Plate can well-grown；The well-grown after 50 DEG C of NA fluid nutrient medium test tube, 200r/min shaking table culture 3d, turbidity are bright
Aobvious, two tests prove that thermoactinomyces sacchari CP can tolerate pH range in 3-12, pH value tolerance with higher.
The test of 8 thermoactinomyces sacchari of embodiment (Laceyella sacchari) CP ethanol tolerance:
The appropriate bacterium of thermoactinomyces sacchari CP slant strains picking is accessed into 100mL NA fluid nutrient medium, 50 DEG C, 200r/
Min shaking table culture for 24 hours seed liquor, first with 1 ‰ inoculum concentrations access be tuned into respectively alcoholic strength be 1%, 3%, 5%, 7%,
9%, in 11%, 13%, 15%, 20% NA fluid nutrient medium, 50 DEG C, 200r/min culture 3d after be coated with NA plate, observation
Growing state；Then seed liquor accesses to the NA fluid nutrient medium test tube of identical alcohol concentration gradient with 1 ‰ inoculum concentrations again, 50
DEG C, 200r/min shaking table culture 3d, observe turbidity.
Test result: thermoactinomyces sacchari CP 50 DEG C of culture 3d in the NA fluid nutrient medium that alcoholic strength is 1-13% are applied
Cloth NA plate can well-grown；The well-grown after 50 DEG C of NA fluid nutrient medium test tube, 200r/min shaking table culture 3d is muddy
Degree is obvious, and it is 1-13%, alcoholic strength with higher that two tests, which prove that thermoactinomyces sacchari CP can tolerate alcoholic strength range,
Tolerance.
The spirit stillage compost maturity test that the prepared by the method spirit stillage decomposing agent of embodiment 9 carries out
1. vinasse compost maturity process
It is with the inoculum concentration access alcoholic strength of mass ratio 1.2 ‰ by spirit stillage decomposing agent prepared by the embodiment of the present invention 1
In the fresh spirit stillage heap body that 10%, pH are 3.58, mix thoroughly, heap body length, width and height are respectively as follows: 200m × 2m × 1.5m, initial heap
Rich water is divided into 65%, when pile temperature is more than or equal to 55 DEG C, starts to carry out first time overturning with dumper, hereafter every 1d overturns one
It is secondary, (the above decomposed processing is defined as processing L)；
The decomposed processing of vinasse heap body of other two same materials, scale is set as control simultaneously, is defined as processing CK1
And CK2, wherein processing CK1 is not to be inoculated with decomposing agent, processing CK2 is to access commercially available spirit stillage decomposing agent, and test method exists together
Manage L.
2. handling L and handling the test result of CK1, processing CK2 comparison:
2.1 temperature
The compost 1d that L is handled it can be seen from attached drawing 1 can reach 55 DEG C, and 3d reaches 85 DEG C of maximum temperature and in 83-
85 DEG C of lasting 3d, can reach compost hazard-free standard；Processing CK2 4d reaches 55 DEG C, and 8d, which reaches 74 DEG C of maximum temperature, only to be tieed up
1d is held, the maximum temperature that processing L reaches is than 11 DEG C of CK2 high of processing；Processing L can terminate decomposed, processing CK2 on the 16th day in fermentation
Terminate after 42d decomposed, relatively processing CK2 shifted to an earlier date 26d and reached decomposed processing L decomposed time phase, handled CK1 in the decomposed ability of 57d
Terminate；The above results show that the degradation of organic matter in spirit stillage heap body can be accelerated by being inoculated with decomposing agent, relative to commercially available white wine wine
Poor decomposing agent, fermentation rate is high when adding spirit stillage quick composting agent compost maturity prepared by the present invention, heating rate is fast, into
The time for entering the megathermal period is short, maintains that the time of megathermal period is long, compost maturity fermentation period is short, have it is significant quickly, it is directly rotten
The characteristics of ripe fresh spirit stillage, without dropping the pretreatment such as alcoholic strength, deacidification.
2.2pH value
It can be seen from attached drawing 2 in entire composting process, processing CK1, processing CK2 and processing L compost later period pH difference
It is 7.74,7.62,7.42, on the whole, inoculation decomposing agent is unobvious on heap body pH influence, but handles the relatively low pH value energy of L
The volatilization for reducing ammonia in composting process, to reduce the loss of nitrogen.
2.3 the content of organic matter
The content of organic matter of processing CK1, processing CK2 and processing L are respectively at the end of compost it can be seen from attached drawing 3
61.28%, 51.75% and 45.85%, reduce 27.15% respectively than the initial content of organic matter (88.43%), 36.68%,
42.58%；Changed by the compost content of organic matter it is found that, the Er Qiejie larger to the degradation of compost organic matter that be inoculated with decomposing agent
The degradation efficiency of kind spirit stillage quick composting agent prepared by the present invention is significantly higher than commercially available spirit stillage decomposing agent.
2.4 total nitrogen content
Three processing are compared it can be seen from attached drawing 4, and the heap body total nitrogen content for being inoculated with decomposing agent is apparently higher than control group
CK1, and it is more slightly higher than the total nitrogen content of commercially available spirit stillage decomposing agent to be inoculated with spirit stillage quick composting agent prepared by the present invention.This
When processing L total nitrogen content be 2.23%, the total nitrogen content for handling CK2 is 2.12%, and the total nitrogen content for handling CK1 is
2.06%.
2.5 moisture content
At the end of compost maturity it can be seen from attached drawing 5, the moisture content of processing L, CK2, CK1 is respectively 21.16%,
23.62%, 25.32%, the content of processing L moisture, which is substantially less than, handles CK1 and processing CK2, adds white wine prepared by the present invention
Fermentation rate is high when vinasse quick composting agent compost maturity, heating rate is fast, into the megathermal period time it is short, maintain the megathermal period
Time is long, effectively accelerates the volatilization of heap body moisture, accelerates decomposed degree, the moisture that can effectively shorten compared with commercially available decomposing agent is waved
The time is sent out, while obtaining the higher lees organic fertilizer of quality.
It should be noted that the spirit stillage decomposing agent of 2-6 of embodiment of the present invention preparation equally has above-mentioned test effect,
Otherness is not significant between each embodiment.
Claims (10)
1. a kind of spirit stillage decomposing agent is mainly prepared by the raw material of following parts by weight: 1-10 parts of thermoactinomyces sacchari microbial inoculum,
1-5 parts of bacillus licheniformis agent, 1-5 parts of bacillus subtilis microbial agent, 1-5 parts of bacillus amyloliquefaciens microbial inoculum；
The thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, deposit number CGMCC
NO.13928。
2. spirit stillage decomposing agent as described in claim 1, which is characterized in that the spirit stillage decomposing agent, mainly by with
It is prepared by the raw material of lower parts by weight: 3-7 parts of thermoactinomyces sacchari microbial inoculum, 2-4 parts of bacillus licheniformis agent, and bacillus subtilis
2-4 parts of microbial inoculum, 2-4 parts of bacillus amyloliquefaciens microbial inoculum.
3. spirit stillage decomposing agent as described in claim 1, which is characterized in that the spirit stillage decomposing agent, mainly by with
It is prepared by the raw material of lower parts by weight: 5 parts of thermoactinomyces sacchari microbial inoculum, 3 parts of bacillus licheniformis agent, and bacillus subtilis microbial agent
3 parts, 3 parts of bacillus amyloliquefaciens microbial inoculum.
4. spirit stillage decomposing agent a method according to any one of claims 1-3, which is characterized in that the thermoactinomyces sacchari microbial inoculum
Preparation method includes the following steps: thermoactinomyces sacchari CP slant strains being inoculated in NA seed bottle solid culture with transfer needle
Base, 60 DEG C of cultures for 24 hours, and thallus are scraped with sterile water elution, and adjustment cell concentration is 5 × 1011Cfu/mL draws 100mL
Bacterium solution is inoculated into 1kg solid medium A, 60 DEG C of culture 48h, dry under the conditions of 60 DEG C to be crushed for 24 hours with pulverizer,
40 meshes are crossed up to thermoactinomyces sacchari microbial inoculum；
The NA seed bottle solid medium the preparation method comprises the following steps: take peptone 10g, powdered beef 3g, sodium chloride 10g, agar
20g, distilled water 1L are uniformly mixed, and are sufficiently dissolved, and adjustment pH value is 7.0, are dispensed into seed bottle after boiling, every bottle of 80-
100mL, 121 DEG C of sterilizing 30min, is put into inclined-plane to obtain the final product；
Solid medium A's the preparation method comprises the following steps: take rice husk 100g, wheat bran 300g, corn flour 200g, beancake powder 100g form sediment
Powder 100g, laterite 100g, calcium sulfate 5g, calcium oxide 7.5g, dipotassium hydrogen phosphate 3g, magnesium sulfate 2g, water 400mL are uniformly mixed,
121 DEG C of sterilizing 45min to obtain the final product.
5. spirit stillage decomposing agent a method according to any one of claims 1-3, which is characterized in that the bacillus licheniformis is lichens
Bacillus ATCC 27811, bacillus licheniformis CICC 10037, bacillus licheniformis CICC 10181, bacillus licheniformis
Any one or a few in CICC 10831, bacillus licheniformis CICC 10291.
6. spirit stillage decomposing agent a method according to any one of claims 1-3, which is characterized in that the bacillus subtilis is withered grass
Bacillus ATCC 6051, bacillus subtilis CICC 10088, bacillus subtilis CICC 10090, bacillus subtilis
Any one or a few in CICC 10153, bacillus subtilis CICC 10157.
7. spirit stillage decomposing agent a method according to any one of claims 1-3, which is characterized in that the bacillus amyloliquefaciens are solution
Bacillus amyloliquefaciens CICC 10035, bacillus amyloliquefaciens CICC 10036, bacillus amyloliquefaciens CICC 10074, Xie Dian
Any one or a few in afnyloliquefaciens CICC 10163, bacillus amyloliquefaciens CICC 20027.
8. spirit stillage decomposing agent a method according to any one of claims 1-3, which is characterized in that the thermoactinomyces sacchari microbial inoculum
Viable bacteria content >=5 × 1011cfu/g；Viable bacteria content >=3 × 10 of the bacillus licheniformis agent11cfu/g；The withered grass bud
Spore bacillus microbial inoculum viable bacteria content >=4 × 1010cfu/g；Viable bacteria content >=3 × 10 of the bacillus amyloliquefaciens microbial inoculum11cfu/
g。
9. the preparation method of spirit stillage decomposing agent a method as claimed in any one of claims 1-8, which is characterized in that including walking as follows
It is rapid: according to formula, precise thermoactinomyces sacchari microbial inoculum, bacillus licheniformis agent, bacillus subtilis microbial agent, solution starch
Gemma bacillus agent is uniformly mixed according to equal increments method up to spirit stillage decomposing agent.
10. application of the spirit stillage decomposing agent a method as claimed in any one of claims 1-8 in terms of quick composting spirit stillage.
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