CN109136148A - A kind of high-temperature fibre element degradation bacillus screening technique and application method - Google Patents
A kind of high-temperature fibre element degradation bacillus screening technique and application method Download PDFInfo
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- CN109136148A CN109136148A CN201811106586.4A CN201811106586A CN109136148A CN 109136148 A CN109136148 A CN 109136148A CN 201811106586 A CN201811106586 A CN 201811106586A CN 109136148 A CN109136148 A CN 109136148A
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- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D1/00—Fertilisers containing potassium
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- C05F11/00—Other organic fertilisers
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention discloses a kind of high-temperature fibre element degradation bacillus screening techniques, using the pig manure sawdust heap of spontaneous fermentation as bacterium source sample, high temperature enriched medium is added, it is screened after enrichment with Congo red selective plate high temperature, the degradation biggish bacterial strain of diameter/colony diameter high-temperature cultivation again is selected, using filter paper item as cellulose degradation indicant, when indicant is disintegrated in culture medium, efficient high temperature fiber element degradation bacteria can be obtained, finally purified.By the above method, the stable high-temperature fibre element degradation bacillus of performance can be effectively screened.The invention also discloses a kind of high-temperature fibre element degradation bacillus application methods, may be directly applied to pig manure fermentation bed after microbial inoculum is made, are convenient for industrialized implementation, have wide application prospect.
Description
Technical field
The present invention relates to agricultural biological technical fields, more particularly, to a kind of degradation bacillus screening of high-temperature fibre element
Method and application method.
Background technique
Fermentation bed to raise pig technology is according to Tiny ecosystem principle and biological fermentation theory, using microorganism to Animal fecal pollution " original
Potential drop solution " reaches the Novel cultivation mode of ecological environment " no pollution ".The technology at first Japan occur and the mouth that puts it over,
Quick promotion and application are obtained in China in recent years.The key of fermentation bed cultivation technology is to support bed, wherein to the science of padding
Maintenance seems with management to be even more important, and well-regulated padding facilitates the digestion and decomposition of feces of livestock and poultry, moreover it is possible to extend padding
Validity period improve culture efficiency.
But when being currently used for the microbial inoculum in fermentation bed for pig raising and being applied on pig manure padding, will receive the antagonism of indigenous bacterium,
It is unfavorable for the decomposed of padding, in addition, existing microbial inoculum is limited to high temperature tolerance ability, will affect heap fertilizer efficiency if fermenting greater than 60 DEG C
Fruit.
Few researchs filter out high temperature fiber element degradation bacteria strains from the protist in pig manure fermentation bed habitat now.With
Pig manure padding is such bacterium of bacterium source screening because deriving from the indigenous bacterium of pig manure padding, and pig manure pad is applied to after microbial inoculum is made
When on material, the antagonism of indigenous bacterium not will receive, thus be more advantageous to the decomposed of padding.To sum up, the efficient fibre of pig manure original inhabitants is screened
Dimension plain degradation bacteria, especially bud pole bacterium, which utilize cellulose resourceization, to be even more important.
Summary of the invention
Goal of the invention of the invention is to provide for a kind of high-temperature fibre element degradation bacillus screening technique, obtains
High-temperature fibre element degradation bacillus, has stronger degradation capability to cellulose, and product not can cause environmental pollution, most
Height can under 60 DEG C of high temperature growth and breeding, can be made into thermophilic fermentation bed addition microbial inoculum, have application in the fermentation bed field of pig manure
Prospect.
The present invention also provides a kind of application of high-temperature fibre element degradation bacillus in pig manure fermentation bed.
To achieve the goals above, the invention adopts the following technical scheme:
A kind of high-temperature fibre element degradation bacillus screening technique of the invention, specific steps are as follows:
(1) after mixing 20wt% pig manure, 40wt% sawdust, 60wt% husk, in 60 DEG C of temperature, 60% humidity, pH7.5
Under conditions of ferment two months, obtain fermentation pig manure sawdust；
(2) using resulting fermentation pig manure sawdust as bacterium source sample, taking 10g fermentation sawdust to be placed in 100mL with CMC-Na is only
In the enriched medium of one carbon source, 50 DEG C, 150rpm constant temperature incubation obtain pregnant solution；
(3) it takes 100 μ l pregnant solutions to be coated on the Congo red culture medium flat plate of CMC-, is inverted in 50 DEG C of incubators later permanent
Temperature culture, is observed after 5 days, occurs hydrolysis on plate, and the big bacterial strain of selection degradation diameter, which is directly inoculated in, is placed with filter paper
It in the enriched medium of item, is disintegrated to filter paper item in culture medium, as the stronger bacterial strain of degradation capability；
(4) a small amount of bacterial strain of picking is crossed on LB solid medium until obtaining single colonie, then the single colonie of acquisition is existed
Using CMC-Na continuously to cultivate for 6 generations on the enriched medium of sole carbon source, to get pure cultivation strain after colonial morphology is stablized, obtain
The Strain Designation arrived is bacillus subtilis SM-5-1 (Bacillus SubtilisSM-5-1), is protected on July 15th, 2018
It is hidden in China typical culture collection center, deposit number is CCTCC NO:M 2018415.
The present invention using the pig manure sawdust heap of spontaneous fermentation as bacterium source sample, be added high temperature enriched medium, after enrichment with
Congo red selectivity plate high temperature screening selects the degradation biggish bacterial strain of diameter/colony diameter high-temperature cultivation again, with filter paper item
Efficient high temperature fiber element degradation bacteria can be obtained when indicant disintegration in culture medium for cellulose degradation indicant, it is most laggard
Row purifying.By the above method, the stable high-temperature fibre element degradation bacillus of performance can be effectively screened.
High-temperature fibre element degradation bacillus of the invention not only has preferable degradation capability, and product to cellulose
It not can cause environmental pollution, since it derives from pig manure, as pig manure indigenous strain, can be reduced and make with the inhibition of indigenous bacterium
With compost and fermentation bed for pig manure are provided with higher degradation capability, are provided simultaneously with high temperature resistance, can be made into thermophilic hair
Ferment bed adds microbial inoculum, has wide application prospect in the fermentation bed field of pig manure.
Preferably, on the basis of enriched medium gross mass, enriched medium is by 98%Mandel nutrient solution and 2%
Carboxymethyl cellulose forms, the composition of nutritive salt in Mandel nutrient solution are as follows: 2g/L NaNO3, 1.5g/L K2HPO4, 1.5g/L
CaCl2, 0.3g/L MgSO4, 0.005g/L FeSO4.7H2O, 0.0016g/LMnSO4.H2O, 0.0014g/L ZnSO4.H2O,
0.0005g/L CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
Preferably, the filter paper specification is 1 × 6cm.
Preferably, the bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is in 60 DEG C of conditions
Lower CMCase and FPase activity is respectively 121.18U/mL and 104.14U/mL.
Preferably, -80 DEG C of use of the bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is low
Warm freezing method preservation.
A kind of high-temperature fibre element degradation bacillus application method, specific steps are as follows:
(1) fermentation bed bacteria is prepared
Bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is inoculated in LB liquid medium,
Bacterium solution is seeded to the wheat bran by 87wt%, the rice chaff of 10wt%, 3wt% quick lime, the matrix of the water composition of 70wt% afterwards for 24 hours
In, 50 DEG C of fermentation 2d wear into fine powder after discharging drying to get hot fermentation bed microbial inoculum.
(2) it ferments
Sawdust is mixed with rice chaff 4:6 in mass ratio first, obtains padding；Pig manure is added in padding again, one cube of padding adds
8kg obtains windrow after adding water adjustment moisture content to be 50~60%；The height of windrow quality 0.5~1% is finally added in windrow
Warm fermentation bed bacteria is completed to ferment for the first time when heap temperature reaches peak and gradually decreases down 45 DEG C, then be turned within every 5-6 days
Heap 1 time, turning 3 times, obtains biological organic fertilizer altogether.
Therefore, the invention has the following beneficial effects:
(1) a kind of high-temperature fibre element degradation bacillus screening technique is provided, obtained high-temperature fibre element degradation
Bacillus has stronger degradation capability to cellulose, and product not can cause environmental pollution, and highest can be under 60 DEG C of high temperature
Growth and breeding can be made into thermophilic fermentation bed addition microbial inoculum, have wide application prospect in the fermentation bed field of pig manure；
(2) a kind of high-temperature fibre element degradation bacillus application method is provided, step is simple, real convenient for industrialization
It applies.
Detailed description of the invention
Fig. 1 is 1 bacillus subtilis SM-5-1 of embodiment (Bacillus Subtilis SM-5-1) culture aspect graph.
Fig. 2 is the aspect graph that experiment is disintegrated in embodiment 1.
Specific embodiment
The present invention will be further described with reference to the accompanying drawings and detailed description.
Embodiment 1
(1) bacterial screening
(1) after mixing 20wt% pig manure, 40wt% sawdust, 60wt% husk, in 60 DEG C of temperature, 60% humidity, pH7.5
Under conditions of ferment two months, obtain fermentation pig manure sawdust；
(2) using resulting fermentation pig manure sawdust as bacterium source sample, taking 10g fermentation sawdust to be placed in 100mL with CMC-Na is only
In the enriched medium of one carbon source, 50 DEG C, 150rpm constant temperature incubation obtain pregnant solution, rich on the basis of enriched medium gross mass
Collection culture medium is made of the carboxymethyl cellulose of 98%Mandel nutrient solution and 2%, the composition of nutritive salt in Mandel nutrient solution
Are as follows: 2g/L NaNO3, 1.5g/L K2HPO4, 1.5g/LCaCl2, 0.3g/L MgSO4, 0.005g/L FeSO4.7H2O,
0.0016g/L MnSO4.H2O, 0.0014g/LZnSO4.H2O, 0.0005g/L CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
(3) it takes 100 μ l pregnant solutions to be coated on the Congo red culture medium flat plate of CMC-, is inverted in 50 DEG C of incubators later permanent
Temperature culture, is observed after 5 days, occurs hydrolysis on plate, and the big bacterial strain of selection degradation diameter, which is directly inoculated in, is placed with specification
It in enriched medium for 1 × 6cm filter paper item, is disintegrated to filter paper item in culture medium, as the stronger bacterial strain of degradation capability, with richness
On the basis of collecting culture medium gross mass, enriched medium is made of the carboxymethyl cellulose of 98%Mandel nutrient solution and 2%,
The composition of nutritive salt in Mandel nutrient solution are as follows: 2g/L NaNO3, 1.5g/L K2HPO4, 1.5g/L CaCl2, 0.3g/L
MgSO4, 0.005g/LFeSO4.7H2O, 0.0016g/L MnSO4.H2O, 0.0014g/L ZnSO4.H2O, 0.0005g/L
CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
(4) a small amount of bacterial strain of picking is crossed on LB solid medium until obtaining single colonie, then the single colonie of acquisition is existed
Using CMC-Na continuously to cultivate for 6 generations on the enriched medium of sole carbon source, to get pure cultivation strain after colonial morphology is stablized, obtain
The Strain Designation arrived is bacillus subtilis SM-5-1 (Bacillus SubtilisSM-5-1), is protected on July 15th, 2018
It is hidden in China typical culture collection center, deposit number is CCTCC NO:M 2018415, is with enriched medium gross mass
Benchmark, enriched medium are made of the carboxymethyl cellulose of 98%Mandel nutrient solution and 2%, nutritive salt in Mandel nutrient solution
Composition are as follows: 2g/LNaNO3, 1.5g/L K2HPO4, 1.5g/L CaCl2, 0.3g/L MgSO4, 0.005g/L FeSO4.7H2O,
0.0016g/L MnSO4.H2O, 0.0014g/L ZnSO4.H2O, 0.0005g/L CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
(2) filter paper slaking test
It is 1 × 6cm filter paper item that two panels specification is placed in enriched medium, is directly inoculated with this on a piece of filter paper item wherein
Invention bacterial strain, the filter paper item disintegration for being inoculated with bacterial strain fragmentate (see Fig. 2), wherein rich on the basis of enriched medium gross mass
Collection culture medium is made of the carboxymethyl cellulose of 98%Mandel nutrient solution and 2%, the composition of nutritive salt in Mandel nutrient solution
Are as follows: 2g/L NaNO3, 1.5g/L K2HPO4, 1.5g/L CaCl2, 0.3g/L MgSO4, 0.005g/L FeSO4.7H2O,
0.0016g/L MnSO4.H2O, 0.0014g/LZnSO4.H2O, 0.0005g/L CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
Illustrate that bacterial strain of the invention has preferable degradation capability to cellulose.
(3) strain idenfication
The 16srDNA of resulting bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is as follows:
AAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCG
GGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGA
TCCGAACTGAGAACAGATTTGTGGGATTGGCTTAGCCTCGCGGCTTCGCTGCCCTTTGTTCTGCCCATTGTAGCACG
TGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACC
TTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGA
CACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGAAGCCCTATCTCTAGGGTTGTCAGAGG
ATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCA
ATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTTGCTGCAGCACTAAAGGGCG
GAAACCCTCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCT
TTCGCGCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCAC
CGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGG
GGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTAC
GTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTACCGCCCTATTCGAAC
GGTACTTGTTCTTCCCTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGAC
TTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGA
TCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCC
ATCTGCAAGTGGTAGCTAAAAGCCACCTTTTATGATTGAACCATGCGGTTCAATCAAGCATCCGGTATTAGCCCCGG
TTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTGACCTAAGGGAGCA
AGCTCCCGTCGGTCCGCTCGACT
The 16SrDNA sequence of bacillus subtilis 5-1 is as shown in SEQ ID No.1.
It is sequenced by 16srDNA, identifies that the bacterial strain is bacillus subtilis (Bacillus Subtilis), be named as withered
Careless bacillus SM-5-1 (Bacillus Subtilis SM-5-1) has been preserved in Chinese Typical Representative training on July 15th, 2018
Object collection is supported, deposit number is CCTCC NO:M 2018415.
(4) measurement of cellulase-producing, filter paper enzyme activity
The thick enzyme supernatant of 0.2ml, 50 DEG C of heat preservation 50min are added in the 1%CMC solution of 1.8ml.It is added in reaction solution
2ml DNS, boiling water bath 10min, ice water are cooling.It is control with inactivator, surveys the absorbance at 550nm.
Reaction system is citrate buffer solution (pH 4.8) 1.4ml, and filter paper considers 0.1g to be worth doing, after 50 DEG C of preheating 10min, addition
0.6ml crude enzyme liquid, 50 DEG C of heat preservation 50min.1ml reaction solution is taken, is added in the reaction tube of the liquid of DNS containing 2ml, boiling water bath 10min,
Ice water terminates reaction.It is control with inactivator, surveys the absorbance at 550nm.
Under the above conditions, the enzyme amount that catalyzing cellulose hydrolysis generates 1 μm of ol glucose per hour is an enzyme activity force
For IU.Enzyme activity calculation formula are as follows: enzyme activity unit (U)=G × n × 103/ m × t × M (glucose μm ol/mlh50
℃)
After measured, bud pole bacterium CMCase and FPase activity under conditions of 60 DEG C be respectively 121.18U/mL and
104.14U/mL。
(5) concrete application method
(1) fermentation bed bacteria is prepared
Bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is inoculated in LB liquid medium,
Bacterium solution is seeded to the wheat bran by 87wt%, the rice chaff of 10wt%, 3wt% quick lime, the matrix of the water composition of 70wt% afterwards for 24 hours
In, 50 DEG C of fermentation 2d wear into fine powder after discharging drying to get hot fermentation bed microbial inoculum.
(2) it ferments
Sawdust is mixed with rice chaff 4:6 in mass ratio first, obtains padding；Pig manure is added in padding again, one cube of padding adds
8kg obtains windrow after adding water adjustment moisture content to be 50~60%；The height of windrow quality 0.5~1% is finally added in windrow
Warm fermentation bed bacteria is completed to ferment for the first time when heap temperature reaches peak and gradually decreases down 45 DEG C, then be turned within every 5-6 days
Heap 1 time, turning 3 times, obtains biological organic fertilizer altogether.
Above-mentioned embodiment is only a preferred solution of the present invention, not the present invention is made in any form
Limitation, there are also other variations and modifications on the premise of not exceeding the technical scheme recorded in the claims.
Sequence table
<110>Zhejiang Academy of Agricultural Science
<120>a kind of high-temperature fibre element degradation bacillus screening technique and application method
<130> 2018
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1401
<212> DNA
<213>bacillus subtilis (Bacillus Subtilis)
<400> 1
aaaggttacc tcaccgactt cgggtgttac aaactctcgt ggtgtgacgg gcggtgtgta 60
caaggcccgg gaacgtattc accgcggcat gctgatccgc gattactagc gattccagct 120
tcacgcagtc gagttgcaga ctgcgatccg aactgagaac agatttgtgg gattggctta 180
gcctcgcggc ttcgctgccc tttgttctgc ccattgtagc acgtgtgtag cccaggtcat 240
aaggggcatg atgatttgac gtcatcccca ccttcctccg gtttgtcacc ggcagtcacc 300
ttagagtgcc caactgaatg ctggcaacta agatcaaggg ttgcgctcgt tgcgggactt 360
aacccaacat ctcacgacac gagctgacga caaccatgca ccacctgtca ctctgccccc 420
gaaggggaag ccctatctct agggttgtca gaggatgtca agacctggta aggttcttcg 480
cgttgcttcg aattaaacca catgctccac cgcttgtgcg ggcccccgtc aattcctttg 540
agtttcagtc ttgcgaccgt actccccagg cggagtgctt aatgcgtttg ctgcagcact 600
aaagggcgga aaccctctaa cacttagcac tcatcgttta cggcgtggac taccagggta 660
tctaatcctg ttcgctcccc acgctttcgc gcctcagcgt cagttacaga ccagagagtc 720
gccttcgcca ctggtgttcc tccacatctc tacgcatttc accgctacac gtggaattcc 780
actctcctct tctgcactca agttccccag tttccaatga ccctccccgg ttgagccggg 840
ggctttcaca tcagacttaa gaaaccgcct gcgcgcgctt tacgcccaat aattccggac 900
aacgcttgcc acctacgtat taccgcggct gctggcacgt agttagccgt ggctttctgg 960
ttaggtaccg tcaaggtacc gccctattcg aacggtactt gttcttccct aacaacagag 1020
ttttacgatc cgaaaacctt catcactcac gcggcgttgc tccgtcagac tttcgtccat 1080
tgcggaagat tccctactgc tgcctcccgt aggagtctgg gccgtgtctc agtcccagtg 1140
tggccgatca ccctctcagg tcggctacgc atcgtcgcct tggtgagccg ttacctcacc 1200
aactagctaa tgcgccgcgg gtccatctgc aagtggtagc taaaagccac cttttatgat 1260
tgaaccatgc ggttcaatca agcatccggt attagccccg gtttcccgga gttatcccag 1320
tcttacaggc aggttaccca cgtgttactc acccgtccgc cgctgaccta agggagcaag 1380
ctcccgtcgg tccgctcgac t 1401
Claims (6)
- A kind of bacillus screening technique 1. high-temperature fibre element is degraded, which is characterized in that specific steps are as follows:(1) by 20wt% pig manure, 40wt% sawdust, 60wt% husk mix after, 60 DEG C of temperature, 60% humidity, pH7.5 item It ferments two months under part, obtains fermentation pig manure sawdust；(2) using resulting fermentation pig manure sawdust as bacterium source sample, 10g fermentation sawdust is taken to be placed in 100mL using CMC-Na as sole carbon In the enriched medium in source, 50 DEG C, 150rpm constant temperature incubation obtain pregnant solution；(3) it takes 100 μ l pregnant solutions to be coated on the Congo red culture medium flat plate of CMC-, is inverted constant temperature training in 50 DEG C of incubators later It supports, is observed after 5 days, occur hydrolysis on plate, the big bacterial strain of selection degradation diameter, which is directly inoculated in, is placed with filter paper item It in enriched medium, is disintegrated to filter paper item in culture medium, as the stronger bacterial strain of degradation capability；(4) a small amount of bacterial strain of picking cross on LB solid medium until obtain single colonie, then by the single colonie of acquisition with CMC-Na, to get pure cultivation strain after colonial morphology is stablized, obtains continuously to cultivate for 6 generations on the enriched medium of sole carbon source Strain Designation be bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1), on July 15th, 2018 protect It is hidden in China typical culture collection center, deposit number is CCTCC NO:M 2018415.
- The bacillus screening technique 2. a kind of high-temperature fibre element according to claim 1 is degraded, which is characterized in that with richness On the basis of collecting culture medium gross mass, enriched medium is made of the carboxymethyl cellulose of 98%Mandel nutrient solution and 2%, The composition of nutritive salt in Mandel nutrient solution are as follows: 2g/L NaNO3, 1.5g/L K2HPO4, 1.5g/L CaCl2, 0.3g/L MgSO4, 0.005g/L FeSO4.7H2O, 0.0016g/L MnSO4.H2O, 0.0014g/L ZnSO4.H2O, 0.0005g/L CoCl2, pH5.5；121 DEG C, 20min, sterilizing.
- The bacillus screening technique 3. a kind of high-temperature fibre element according to claim 1 is degraded, which is characterized in that described Filter paper specification is 1 × 6cm.
- The bacillus screening technique 4. a kind of high-temperature fibre element according to claim 1 is degraded, which is characterized in that described Bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) CMCase and FPase activity point under conditions of 60 DEG C It Wei not 121.18U/mL and 104.14U/mL.
- The bacillus screening technique 5. a kind of high-temperature fibre element according to claim 1 is degraded, which is characterized in that described Bacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) -80 DEG C of cryogenic freezing preserving process preservations of use.
- A kind of bacillus application method 6. high-temperature fibre element is degraded, which is characterized in that specific steps are as follows:(1) fermentation bed bacteria is preparedBacillus subtilis SM-5-1 (Bacillus Subtilis SM-5-1) is inoculated in LB liquid medium, for 24 hours after Bacterium solution is seeded to the wheat bran by 87wt%, the rice chaff of 10wt%, 3wt% quick lime, in the matrix of the water composition of 70wt%, 50 DEG C fermentation 2d, discharging drying after wear into fine powder to get hot fermentation bed microbial inoculum；(2) it fermentsSawdust is mixed with rice chaff 4:6 in mass ratio first, obtains padding；Pig manure is added in padding again, one cube of padding adds 8kg, Windrow is obtained after adding water adjustment moisture content to be 50~60%；The high temperature hair of windrow quality 0.5~1% is finally added in windrow Ferment bed microbial inoculum is completed to ferment for the first time when heap temperature reaches peak and gradually decreases down 45 DEG C, then every turning in 5-6 days 1 Secondary, turning 3 times, obtains biological organic fertilizer altogether.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110964639A (en) * | 2019-12-09 | 2020-04-07 | 湖南金泥生物科技有限公司 | Strain screening method applied to pig raising fermentation bed and application |
| CN114134077A (en) * | 2021-11-19 | 2022-03-04 | 江苏科技大学 | Silkworm excrement-derived cellulose degrading bacterium DC11 and screening method and application thereof |
| CN114685213A (en) * | 2020-12-30 | 2022-07-01 | 北京农学院 | Preparation method of organic compound fertilizer for peach branches |
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| CN101525583A (en) * | 2008-09-18 | 2009-09-09 | 华中农业大学 | Bacillus subtilis dcy-1 and application thereof in biofermentation |
| CN106350469A (en) * | 2016-11-09 | 2017-01-25 | 上海交通大学 | Bacillus with high temperature resistance and cellulose degradation capacity and application thereof |
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| CN101525583A (en) * | 2008-09-18 | 2009-09-09 | 华中农业大学 | Bacillus subtilis dcy-1 and application thereof in biofermentation |
| CN106350469A (en) * | 2016-11-09 | 2017-01-25 | 上海交通大学 | Bacillus with high temperature resistance and cellulose degradation capacity and application thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110964639A (en) * | 2019-12-09 | 2020-04-07 | 湖南金泥生物科技有限公司 | Strain screening method applied to pig raising fermentation bed and application |
| CN114685213A (en) * | 2020-12-30 | 2022-07-01 | 北京农学院 | Preparation method of organic compound fertilizer for peach branches |
| CN114134077A (en) * | 2021-11-19 | 2022-03-04 | 江苏科技大学 | Silkworm excrement-derived cellulose degrading bacterium DC11 and screening method and application thereof |
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