CN107988099A - A kind of microbial bacterial agent of organic waste fast degradation minimizing and its application - Google Patents
A kind of microbial bacterial agent of organic waste fast degradation minimizing and its application Download PDFInfo
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- CN107988099A CN107988099A CN201711260594.XA CN201711260594A CN107988099A CN 107988099 A CN107988099 A CN 107988099A CN 201711260594 A CN201711260594 A CN 201711260594A CN 107988099 A CN107988099 A CN 107988099A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/20—Bacteria; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F9/00—Fertilisers from household or town refuse
- C05F9/04—Biological compost
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The present invention provides microbial bacterial agent and its application of a kind of organic waste fast degradation minimizing, and microbial inoculum is mixed by bacillus subtilis (Bacillus subtilis) that preserving number is CGMCC No.14052, the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) of preserving number CGMCC No.14053, the bacillus subtilis (Bacillus subtilis) that preserving number is CICC No.10210 and cellulose-degrading bacteria (Bacillus sp) that preserving number is CICC No.10829.The microbial bacterial agent is mixed with carrier rice chaff, microbial bacterial agent is attached in carrier rice chaff；Press filtration is handled after organic waste is sieved；Organic waste after press filtration is handled is added in reaction warehouse, adds the carrier rice chaff of attached microbial microbial inoculum, and intermittent stirring reacts 12~24h, and more than 90% decrement can be reached to kitchen garbage.Present invention treatment in situ organic waste from source, greatly alleviates the pressure of organic garbage treatment.
Description
Technical field
The present invention relates to the technical field of a kind of organic waste microorganism fast degradation minimizing, harmless treatment, specifically
It is related to screening, organic waste minimizing, the control of harmless treatment condition of degraded organic waste high-effective microorganism microbial inoculum.
Background technology
Organic waste is " wet refuse ", with the quickening of economic fast development and urbanization, living standards of the people
Improve, organic waste yield is increasing, and kitchen garbage accounts for the 50-70% of organic waste, and kitchen garbage yield is got over
Come bigger, become increasingly complex.Statistics shows that China city produces kitchen garbage more than 60,000,000 tons every year, daily more than 16
Ten thousand tons.But from the point of view of current preceding 5 batches of 100 experimental cities, most project is built up also in building, establishing the stage
It is less, normal operation it is less.
For organic waste since water content is high, caloric value is low, is such as burned with other house refuses, it is impossible to meets that rubbish is burnt
The caloric value requirement of power generation is burnt, and the harmful toxic matters such as bioxin can also be produced and endanger people to health.Its component is main
Sugar, protein, starch, fat, cellulose, the content of organic matter is high, full of nutrition, perishable rotten, if take with it is other
Domestic garbage mixing carries out sanitary landfills, easily causes stench, produces a large amount of percolates and biogas, contaminated soil, water source and big
Gas, endangers people's health.The organic waste content of organic matter is high, and recycle value is high, if lost with consumer waste filling and embedding
Utility value.Based on considerations above, carry out organic waste and be collected separately and handle necessary, and urban modernization management
Inexorable trend.
The recycling of organic waste, minimizing, harmless treatment have following social benefit：(1) organic waste is made in source
It is under control on head, promotes the consciousness of resident's garbage classification, is conducive to promote the progress of garbage classification；(2) organic waste is just
Ground is disposed, and is reduced the construction of environmental sanitation clearing and garbage transfer station, is avoided secondary pollution in transit, reduce at rubbish
The cost of reason；(3) food safety hazards such as gutter oil, swill pig are eliminated from root；(4) garbage disposal is reduced to environment
Pollution and the harm to health of people；(5) recycled by disposal of resources, realization.How effectively to handle and dispose and is organic
Rubbish, it is an important topic of urban modernization management to realize recycling, innoxious, minimizing.In recent years, the world is caused
The great attention of national governments.
The current processing method of organic garbage treatment mainly crushes in line, sanitary landfills, compost, high temperature feed, heat
Solution, fermentation and hydrogen production method etc..Since the kitchen garbage oil content in China is high, in line easily blocking pipeline is crushed, is unsuitable for long-term
Operation.Organic waste sanitary landfills is easy to operate, and occupied ground is big, produces a large amount of landfill leachates, brings secondary pollution, European Union
This processing method is forbidden.The shortcomings that organic waste compost is of low quality, and the composting mode of better quality is of high cost, is promoted
It is difficult.High-temperature process feed, there are the safety issue of feed, the raising required in face of disinfection, processing cost increase, equipment
Requirement also improve.Take low burning method moisture height, calorific value, generation bioxin welding and endanger people's health.
Although pyrolysismethod has broad application prospects, but of high cost, technology not yet reaches the practical stage.Fermentation and hydrogen production is to generally believe
Most potential alternative energy source, but still conceptual phase in the early stage.Microbial bacterial agent fast degradation processing organic waste operation into
This low, easy to operate, non-secondary pollution, carries out treatment in situ minimizing, innoxious has certain advantage.
The content of the invention
The present invention provides a kind of organic waste microorganism fast degradation minimizing, innoxious processing method, from source
Treatment in situ organic waste, greatly alleviates the pressure of organic garbage treatment.
A kind of microbial bacterial agent of organic waste fast degradation minimizing, is the withered of CGMCC No. 14052 by preserving number
Careless bacillus (Bacillus subtilis), the bacillus amyloliquefaciens that preserving number is CGMCC No.14053
(Bacillus amyloliquefaciens), preserving number are the bacillus subtilis (Bacillus of CICC No.10210
Subtilis) it is mixed with preserving number for the cellulose-degrading bacteria (Bacillus sp) of CICC No.10829.
The microbial bacterial agent of the present invention is acidproof salt tolerant microbial inoculum, and its growth pH is 3.5~6.0, and tolerance NaCl can reach
2.5%, cultivation temperature is 25~45 DEG C.Preferably, by after each bacterial strain individually culture, bacterium night is pressed 1:1:1:1 mixes.
High-effective microorganism microbial inoculum of the present invention, is the feature and main component sieve that acid and high salt are easily produced for organic waste
The efficient degradation starch of choosing, protein, fat, the acidproof degradation bacteria of the salt tolerant of cellulose.
Starch of the present invention, protein degradation bacterium are to screen to obtain from the kitchen garbage treatment fluid tamed for a long time.
It for Bacillus subtilis, bacterial strain number is HKY-1 that starch degrading bacterium, which uses Classification And Nomenclature, preserving number CGMCC
The bacillus subtilis of No.14052.Protein degradation bacterium is that bacillus amyloliquefaciens Classification And Nomenclature is Bacillus
The bacillus amyloliquefaciens that amyloliquefaciens, bacterial strain number are HKY-2, preserving number is CGMCC No.14053.Two plants of bacterium
It is preserved in the China Microbiological bacterium positioned at Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica
Kind preservation administration committee common micro-organisms center, preservation date is on April 20th, 2017.
Fat, the cellulose-degrading bacteria of the present invention is filtered out from the strain that laboratory has preserved, and is from China
Research for Industrial Microbial Germ collection is commercially available, and fat acid decomposition bacterium is bacillus subtilis Bacillus subtilis CICC
No.10210, cellulose-degrading bacteria are Bacillus Bacillus sp CICC No.10829.
After these four bacterium of the present invention filter out, choose single bacterium colony from tablet and be inoculated into LB fluid nutrient mediums and cultivate, its
Condition of culture is that pH is that 3.5~5, NaCl can reach 2.0~2.5%, and cultivation temperature is 25~45 DEG C, and 120~160rpm, shakes
Bed culture 3~6 days can be according to underlying scale 1:1:1:1 ratio mixing adds, can be according to the specific of processing in actual conditions
Component of refuse adjusts microbial inoculum ratio.
Further preferred condition of culture pH is 4.0, NaCl 2.0%, and cultivation temperature is 35 DEG C, 140rpm, shaking table training
Support 4 days.
The present invention also provides a kind of method of organic waste fast degradation minimizing, include the following steps：
(1) microbial bacterial agent is mixed with carrier rice chaff, microbial bacterial agent is attached in carrier rice chaff；By organic rubbish
Press filtration is handled after rubbish screening；
(2) organic waste after press filtration is handled is added in reaction warehouse, adds the carrier rice chaff of attached microbial microbial inoculum,
Intermittent stirring reacts.
The efficient acidproof Salt-tolerant microbial agent of present invention screening degraded organic waste；Using aerobic and facultative microbe pair
Organic waste is degraded, it is general 12~24 it is small when can reach more than 90% solid-state fluorescence material it is degradable for liquid and
CO2；The liquid that microbial degradation organic waste produces is discharged into municipal sewage pipe network after Anaerobic Treatment, is concentrated in sewage plant
Can qualified discharge after processing.
The features such as present invention is for organic waste high salt, Gao Shui, high oily, perishable production acid, exploitation salt tolerant is acidproof efficiently multiple
Microbial bacterial agent is closed, need to only carry out preliminary screening, drainage to organic waste, the bulk such as bone, cornstalk rubbish is carried out must
The crushing wanted, is added in organic garbage disposal reaction warehouse, is uniformly mixed with the rice chaff carrier of attached microbial microbial inoculum, with
Extremely low speed intermittent stirring, after 12~24h, microorganism can be by more than 90% solid by the high-effective microorganism enzyme of generation
Organic waste is degradable into liquid and CO2.The liquid that degraded produces can enter municipal sewage pipe network by Anaerobic Treatment heel row,
It is not only that urban wastewater treatment firm provides good carbon source, and can qualified discharge after sewage plant centralized processing.The technology has
There is the advantages of waste reduction rate is high, specific energy consumption is low, floor space is small, treatment effeciency is high, processing cost is low.For organic rubbish
The distribution of rubbish, especially kitchen garbage dissipates, is distributed the characteristics of wide, amount is few, and domestic rubbish disposal is greatly alleviated from source
Pressure.
It is preferred that using carrier of the rice chaff as microbial bacterial agent, rice chaff, contact area is big, and can ensure the shape of simultaneous oxygen
State.
Preferably, the bulky grain organic waste screened out is put into reaction warehouse after crushed in step (1).Institute of the present invention
After the organic waste stated is, it is necessary to first sort the organic matters of the bulks such as bone and stalk, crushing be added in reaction warehouse into
Row degradation treatment.
Preferably, the organic waste moisture after press filtration processing is controlled 20~40%.Further preferred water content is 25
~35%, it is most preferably 30%.The moisture being press-filtered out enters municipal sewage pipe network after carrying out Anaerobic Treatment, and only sewage plant does not carry
For good carbon source, moreover it is possible to the qualified discharge after sewage plant centralized processing.
Organic waste of the present invention is added in reaction warehouse and is mixed with the rice chaff carrier of attached microbial microbial inoculum, interval
Stirring, ensures that microbial bacterial agent carries out fast degradation under the conditions of aerobic and simultaneous oxygen to organic waste, it is preferable that stirring 5~
10min, is spaced 10~15min, and mixing speed is 5~20rpm, and reaction temperature is 34 DEG C~45 DEG C, and reactant pH is controlled 3.5
~5.5, humidity is 25~35% in reaction warehouse.
Further preferably, 5~6min, 10~12min of interval are preferably stirred；Mixing speed is 8~10rpm；Temperature is 35
DEG C~40 DEG C；PH is controlled 4.0~4.5.
Preferably, for every a batch of organic waste, the time of the interior interval stirring reaction of reaction warehouse is 12~24h.This
The processing method of invention, organic waste pass through 12~24h, and the organic waste for reaching more than 90% is degraded to liquid and CO2。
Preferably, the dosage of microbial bacterial agent is using the mass ratio of day output in microbial bacterial agent and reaction warehouse as 0.3
~0.5% meter, added once at interval of 10~15 days.
Preferably, the liquid produced after step (2) processing enters municipal sewage pipe network through Anaerobic Treatment heel row.It is not only city
Town sewage treatment plant provides good carbon source, and can qualified discharge after sewage plant centralized processing.
Preferably, a small amount of stink produced in step (2) processing procedure is pumped into the equipment equipped with deodorant and purifies heel row
Enter sewage network.Deodorant is the deodorant of commercialization, is a kind of plants essential oil.
Preferably, the organic waste is kitchen garbage.
Compared with prior art, the invention has the advantages that：
(1) degradation speed is fast.The technique uses the acidproof microorganism of salt tolerant, to organic waste under conditions of aerobic and simultaneous oxygen
Degradation speed it is fast compared with fermenting speed, it is general 12~24 it is small when more than 90% organic waste can be completely decomposed into liquid and
CO2.And zymotechnique generally require 24 it is small when or so could complete the fermentation process of organic waste.
(2) catabolite is few.Zymotechnique is compared using the acidproof aerobic treatment process with facultative microbe microbial inoculum of salt tolerant
Degrade to organic waste more thorough, therefore organic waste reduction rate higher, processing product are discharged in liquid form, can make liquid fertilizer
For pouring, also septic tank can be also directly discharged to as the nutrient solution of sewage disposal system biochemistry pool, or after Anaerobic Treatment
Mix with sanitary sewage and discharged after being discharged into sewage disposal system processing, without compost, the processing for significantly reducing catabolite is difficult
Degree.
(3) stink produced is few.Similar product is produced due to using hot fermentation treatment process in processing procedure
A large amount of stinks.The present invention is few using aerobic and facultative microbe treatment process, the stink of generation.
(4) heating is few needed for.Similar product using hot fermentation due to being handled, and stir storehouse temperature general control is 45
, it is necessary to constantly heat to keep temperature in storehouse between~70 DEG C.The present invention is using aerobic and facultative microbe to organic waste
Degraded carry out at normal temperatures, therefore the temperature control of stir storehouse is at 10~50 DEG C, general only minimum in winter temperature
When just need to carry out the degradation efficiency that a small amount of heating ensures organic waste.
(5) power consumption is few.Similar product often handles the power consumption of 1 kilogram of organic waste generally in 0.15~0.20kWh
Between.And this method processing organic waste is used since demand for heat is few, the power consumption of 1 kilogram of organic waste of processing exists
Below 0.10kWh.
Brief description of the drawings
Fig. 1 is starch degrading bacterium bacillus subtilis Bacillus subtilis HKY-1, CGMCC No.14052,
On starch culture-medium tablet, add the transparent circle produced after iodine solution.
Fig. 2 is that protein degradation bacterium is bacillus amyloliquefaciens Bacillus amyloliquefaciens HKY-2,
CGMCC No.14053, the transparent circle produced on casein culture medium flat plate.
Fig. 3 is fat acid decomposition bacterium Bacillus subtilis CICC No.10210, the production fat produced on grease tablet
Fat acid indicator is by bluish-green flavescence.
Fig. 4 is cellulose-degrading bacteria Bacillus Bacillus sp CICC No.10829, Congo red flat in cellulose
The transparent circle produced on plate.
Embodiment
The screening of bacterial strain：
The screening technique of efficient acidproof salt tolerant starch degrading bacterium：Culture medium (g/L)：Soluble starch 5g, peptone 5g,
NaCl 25g, beef extract 5g, pH are adjusted to 4.0, after high-temperature sterilization, are inoculated with kitchen garbage treatment fluid 10g, 37 DEG C, 150rpm shakes
After bed culture domestication enrichment 7 days, inoculation 5ml nutrient solutions to identical culture medium so repeat enrichment 2 times.Take last time
Enrichment culture liquid 1ml, using 10 times of gradient dilution methods, tablet is coated on the solid medium of same medium, 37 DEG C of inversions
Culture, until there is single bacterium colony to grow, adds iodine solution on tablet, observes transparent circle size.The production amylase of laboratory preservation
Bacterial strain is tested together, the larger single bacterium colony of picking transparent circle, line purifying.The single bacterium colony that will be singled out is seeded in same liquid training
Support in base, 150rpm, 37 DEG C of shaking table culture 48h, after taking zymotic fluid to centrifuge, 1ml supernatants are taken, using 3,5- dinitrosalicylic acid systems
The amylase activity size produced is surveyed, filters out one plant of bacterium of enzyme activity maximum, bacterial strain number is denoted as HKY-1, as experiment
Efficient degrading bacteria.Identified using 16S rDNA sequence pair HKY-1 bacterial strains, be sequenced and completed by sequencing company, 16S rDNA sequences are such as
Shown in SEQ ID NO.1, sequence Genbank carry out Blast after find, the 16S rDNA sequences of bacterial strain HKY-1 with
The homology of Bacillus subtilis is 99%.
Confirm that bacterial strain is Bacillus subtilis by identifying, be named as bacillus subtilis Bacillus
Subtilis HKY-1, are preserved in positioned at Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica
China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date are on April 20th, 2017.
The screening technique of efficient acidproof salt tolerant protein matter degradation bacteria：Culture medium (g/L)：Casein 5g, glucose 2g, K2HPO4
0.3g, MgSO40.2g, NaCl 25g, pH are adjusted to 4.0, after high-temperature sterilization, are inoculated with kitchen garbage treatment fluid 10g, 37 DEG C,
After 150rpm shaking table cultures domestication enrichment 7 days, inoculation 5ml nutrient solutions to identical culture medium so repeat enrichment 2 times.Take last
Enrichment culture liquid 1ml once, carries out tablet coating, 37 DEG C are inverted culture, until there is single bacterium colony using 10 times of gradient dilution methods
Grow.The bacteria produced proteinase strain of Laboratories Accession is together tested at the same time and is compared, the larger list of bacterium colony transparent circle on picking tablet
Bacterium colony, line purify to obtain single bacterium colony.The single bacterium colony of different strains is inoculated with carry out liquid fermentation and culture respectively.Fermentation medium：
Wheat bran 15g, analysis for soybean powder 8%, glucose 3%, NaH2PO41%, CuSO40.2%, NaCl 2.0%, pH4.5, most suitable fermentation
Temperature is 37 DEG C, 150rpm, fermented and cultured 2 days.After zymotic fluid centrifugation, 1ml supernatants are taken, albumen is filtered out using folin's methods
One plant of bacterium of enzyme activity maximum is as experiment efficient degrading bacterial strain.
Identified using 16S rDNA sequence pair HKY-2 bacterial strains, be sequenced and completed by sequencing company, 16S rDNA sequences such as SEQ
Shown in ID NO.2, sequence is found after Genbank carries out Blast, the 16S rDNA sequences and Bacillus of bacterial strain HKY-2
The homology of amyloliquefaciens is 99%.
Confirm that bacterial strain is Bacillus amyloliquefaciens by identifying, be named as bacillus subtilis
Bacillus amyloliquefaciens HKY-2, are preserved in positioned at section of Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China
The China Committee for Culture Collection of Microorganisms's common micro-organisms center of institute of microbiology of institute, preservation date are 2017
April 20.
The screening technique of efficient acidproof salt tolerant fat acid decomposition bacterium：Culture medium (g/L)：Dusty yeast 1g, olive oil emulsion
20g, (NH4)2SO42g, NaCl 25g, KH2PO40.15g, Na2HPO40.35g, pH are adjusted to 7.0, after high-temperature sterilization, connect
Kind grease contaminated soil sample 10g, after the domestication of 150rpm shaking table cultures is enriched with 7 days, takes 5ml nutrient solutions to be inoculated into phase by 37 DEG C
Same culture medium so repeats enrichment 2 times.The enrichment culture liquid 1ml of last time is taken, is coated on using 10 times of gradient dilution methods
On same medium tablet containing final concentration of 0.2% elegant thymol blue indicator, 37 DEG C are inverted culture, until there is single bacterium colony length
Go out.The yielding lipase bacterial strain of Laboratories Accession, which is together tested, to be compared, and picking single bacterium colony surrounding media is turned yellow by bluish-green flavescence
The big single bacterium colony of scope is fermented.150rpm, 37 DEG C after fermentation culture for 2 days, takes 1ml zymotic fluids to take supernatant after centrifuging, detects
Lipase activity.
Hexane extraction process surveys lipase activity：1ml olive oil is taken to be placed in containing glycine-NaOH pH9.4 buffer solutions
In 3ml, 1ml fermented supernatant fluids are added, 37 DEG C of reaction 10min, are terminated with 15ml ethanol-acetones and reacted, with 10ml water saturations just
Hexane extraction generation aliphatic acid, from upper strata take 2.5ml to be placed in dry conical flask with phenolphthalein give instruction agent use
The aliphatic acid of 0.025mol/KOH ethanol solutions titration generation.Under the above-described reaction conditions, catalysis per minute is produced into 1 μm of ol fat
The required enzyme amount of fat acid is defined as 1 unit of activity (U).So as to the yield of enzyme size of more every plant of bacterium, it is big to select enzyme activity
Bacterial strain is as test organisms.
The screening technique of highly effective cellulose degradation bacteria：Culture medium (g/L)：Stalk powder 5g, filter paper bar 1g, KH2PO41g,
NH4NO3 1g,MgSO4.7H2O 0.5g, yeast extract 1g, NaCl 25g, pH 5.0, after high-temperature sterilization, is inoculated with 10g straw under high temperature
Compost sample, 150rpm, 37 DEG C of domestications are enriched with 10 days, are taken 5ml nutrient solutions to be inoculated into identical culture medium and are so repeated enrichment 2
It is secondary.The enrichment culture liquid 1ml of last time is taken, the upper of same medium, 37 DEG C of inversions are coated on using 10 times of gradient dilution methods
Culture 10 days, the cellulose-degrading bacteria of Laboratories Accession is together tested, and compares the big of after congo red has been added transparent circle
It is small, choose several plants of big bacterium of generation transparent circle and carry out fermentation measure enzyme activity size.
Fermentation medium (g/L)：Sodium carboxymethylcellulose 15g, KH2PO41g, NH4NO31g, MgSO4.7H2O 0.5g,
Yeast extract 1g, NaCl 25g, pH5.0, after high-temperature sterilization, are inoculated with this several plants of bacterium respectively, 150rpm, 37 DEG C of shaking table cultures 4 days, hair
After zymotic fluid centrifugation, 1ml supernatants are taken to choose enzyme activity maximum using the size of 3,5- dinitrosalicylics acid system measure enzyme activity
One plant of bacterium is as test organisms.
As shown in Figure 1 to 4, wherein Fig. 1 is the transparent circle that 4 kinds of efficient acidproof salt tolerant degradation bacterias produce on tablet
Bacillus subtilis HKY-1 CGMCC No.14052, Fig. 2 are Bacillus amyloliquefaciens HKY-2
CGMCC No.14053；Fig. 3 is Bacillus subtilis CICC No.10210；Fig. 4 is Bacillus sp CICC
No.10829。
Embodiment 1
Hangzhou middle school dining room, about 0.5 ton of kitchen garbage of day processing, treatment conditions are as follows：
Screen four kinds of bacterium are individually cultivated in LB liquid medium, its condition of culture is that pH is that 4.0, NaCl can
To reach 2.0%, cultivation temperature is 35 DEG C, 140rpm, and shaking table culture is after 3~6 days, according to 1:1:1:1 volume ratio mixes
Close.
Mixed microbial inoculum is stirred with rice chaff, not lumpd to be easily added in reaction warehouse.Microbial bacterial agent is per two weeks
Add once, each dosage is 0.4% (M/M) of organic waste day output.
, it is necessary to which the bulk organic waste such as bone, corn block to bulk carries out sorting out crushing before organic waste adds
After be added in reaction warehouse and carry out degradation treatment.
Organic waste needs to carry out it press filtration moisture control 25~35% before adding, filtrate passes through Anaerobic Treatment
Afterwards, municipal sewage pipe network is discharged into, only urban wastewater treatment firm does not provide good carbon source, and after sewage plant centralized processing
Can qualified discharge.
Organic waste after sorting crushing drainage is added in reaction warehouse to be mixed with the rice chaff carrier of attached microbial microbial inoculum,
Intermittent stirring, ensures that microbial bacterial agent carries out fast degradation under the conditions of aerobic and simultaneous oxygen to organic waste, stirs 5min, interval
10min.Mixing speed 10rpm.For reaction warehouse temperature at 36 DEG C~40 DEG C, reactant pH controls are wet in 3.5~5.0, reaction warehouse
Spend for 25~35%.
Organic waste passes through 12~24h, and the organic waste for reaching more than 90% is degraded to liquid and CO2.What degraded produced
Liquid is discharged into municipal sewage pipe network after Anaerobic Treatment, and only urban wastewater treatment firm does not provide good carbon source, Er Qie
Can qualified discharge after sewage plant centralized processing.
Kitchen garbage is perishable because the content of organic matter is high, finds to enable in practice degradable in 12~24h energy
As liquid and CO2, also can slightly a little stinks, it is necessary to which the stink produced using exhaust fan is extracted out, by equipped with deodorant
Equipment in handle after be discharged into sewage network.
Table 1：The result of the test of 0.5 ton of kitchen garbage of day processing
| Processing time | Water content (%) | Temperature (DEG C) | pH | Reduction rate (%) |
| First week | 28.6 | 34.6 | 4.4 | 90.2 |
| Second week | 27.5 | 35.3 | 4.7 | 90.7 |
| 3rd week | 29.6 | 37.6 | 4.8 | 91.4 |
| 4th week | 29.4 | 36.9 | 4.3 | 90.5 |
| 5th week | 30.1 | 38.5 | 4.7 | 90.6 |
| 6th week | 31.4 | 36.6 | 4.7 | 90.5 |
| 7th week | 30.9 | 37.4 | 4.3 | 91.4 |
| 8th week | 32.2 | 36.5 | 4.6 | 91.6 |
Embodiment 2
The kitchen garbage of each eating and drinking establishment of Hangzhou Commercial Complex, about 0.3~0.8 ton of day output, the kitchen garbage
Maximum feature is fat content height, and the characteristics of for the kitchen garbage, treatment conditions are as follows.
Screen four kinds of bacterium are individually cultivated in LB liquid medium, its condition of culture is that pH is that 4.0, NaCl can
To reach 2.0%, cultivation temperature is 35 DEG C, 140rpm, and shaking table culture is after 3~6 days, degradable starch, protein, fat, fiber
The ratio of element is according to 1:1:4:1 volume ratio mixing.
Mixed microbial inoculum is stirred with rice chaff, not lumpd to be easily added in reaction warehouse.Microbial bacterial agent is per two weeks
Add once, each dosage is 0.4% (M/M) of organic waste day output.
, it is necessary to which the bulk organic waste such as bone, corn block to bulk carries out sorting out crushing before organic waste adds
After be added in reaction warehouse and carry out degradation treatment.
Organic waste needs to carry out it press filtration moisture control 25~35% or so before adding, filtrate is by anaerobism
After reason, municipal sewage pipe network is discharged into, only urban wastewater treatment firm does not provide good carbon source, and is focused in sewage plant
After can qualified discharge.
Organic waste after sorting crushing drainage is added in reaction warehouse to be mixed with the rice chaff carrier of attached microbial microbial inoculum,
Intermittent stirring, ensures that microbial bacterial agent carries out fast degradation under the conditions of aerobic and simultaneous oxygen to organic waste, stirs 10min,
Have a rest 15min.Mixing speed 10rpm.At 36 DEG C~40 DEG C, reactant pH is controlled in 3.5~4.5, reaction warehouse reaction warehouse temperature
Humidity is 25~35%.
Organic waste passes through 12~24h, and the organic waste for reaching more than 90% is degraded to liquid and CO2.What degraded produced
Liquid is discharged into municipal sewage pipe network after Anaerobic Treatment, and only urban wastewater treatment firm does not provide good carbon source, Er Qie
Can qualified discharge after sewage plant centralized processing.
Kitchen garbage is perishable because the content of organic matter is high, finds to enable in practice degradable in 12~24h energy
As liquid and CO2, also can slightly a little stinks, it is necessary to which the stink produced using exhaust fan is extracted out, by equipped with deodorant
Equipment in handle after be discharged into sewage network.
Table 2：The result of the test of 0.3~0.8 ton of kitchen garbage of day processing
| Processing time | Water content (%) | Temperature (DEG C) | pH | Reduction rate (%) |
| First week | 29.6 | 36.8 | 4.5 | 91.4 |
| Second week | 31.3 | 37.9 | 4.3 | 90.3 |
| 3rd week | 28.9 | 36.4 | 4.7 | 91.9 |
| 4th week | 27.4 | 38.9 | 4.9 | 90.8 |
| 5th week | 30.7 | 38.2 | 4.6 | 89.5 |
| 6th week | 29.3 | 37.4 | 4.3 | 90.6 |
| 7th week | 31.6 | 38.3 | 4.9 | 91.2 |
| 8th week | 27.8 | 36.6 | 4.1 | 89.4 |
Embodiment 3
The vegetable cultivation base in Zhejiang Ruian village, 1~1.5 ton of abandoned vegetable of day output, abandoned vegetable 1 year is with season
The change process component of section is varied from, and spring and summer is mainly common green vegetable, and autumn contains bamboo shoot leaf, maize straw etc., place
Manage bar part is as follows.
Screen four kinds of bacterium are individually cultivated in LB liquid medium, its condition of culture is that pH is that 4.0, NaCl can
To reach 2.0%, cultivation temperature is 35 DEG C, 140rpm, and shaking table culture is after 3~6 days, degradable starch, protein, fat, fiber
The bacterium of element is according to 1:1:1:6 volume ratio mixing.
Mixed microbial inoculum is stirred with rice chaff, not lumpd to be easily added in reaction warehouse.Microbial bacterial agent is per two weeks
Add once, each dosage is 0.4% (M/M) of organic waste day output.
, it is necessary to carry out crushing the reaction warehouse added to the bulk such as maize straw, dried bamboo shoots organic waste before organic waste adds
Interior carry out degradation treatment.
The organic wastes such as vegetables are added in reaction warehouse and are mixed with the rice chaff carrier of attached microbial microbial inoculum, intermittent stirring,
Ensure that microbial bacterial agent carries out fast degradation under the conditions of aerobic and simultaneous oxygen to organic waste, stir 10min, interval 10min.Stir
Mix speed 10rpm.Reaction warehouse temperature is at 36 DEG C~40 DEG C, and humidity is 25~30% in reaction warehouse, reactant pH controls 4.5~
5.5。
Organic waste passes through 12~24h, and the organic waste for reaching more than 90% is degraded to liquid and CO2。
For can degradable be liquid in the basic 12h of organic waste as fresh abandoned vegetable one kind, bamboo shoot leaf and
Generally reach after corn straw smashing in 24 more than 90% it is degradable be liquid and CO2, so not producing substantially any smelly
Taste, it is not necessary to add deodorant deodorization.The liquid that degraded produces can be as the high-quality organic fertilizer in vegetable cultivation base.
Table 3：The result of the test of 1~1.5 ton of abandoned vegetable of day processing
| Processing time | Water content (%) | Temperature (DEG C) | pH | Reduction rate (%) |
| First week | 30.4 | 38.4 | 5.4 | 95.4 |
| Second week | 28.4 | 36.4 | 5.7 | 96.3 |
| 3rd week | 32.6 | 35.3 | 5.8 | 97.9 |
| 4th week | 26.5 | 37.5 | 6.0 | 93.8 |
| 5th week | 32.8 | 38.9 | 5.3 | 90.5 |
| 6th week | 25.3 | 35.8 | 5.7 | 90.6 |
| 7th week | 23.6 | 36.9 | 6.2 | 93.2 |
| 8th week | 30.3 | 35.4 | 5.9 | 92.6 |
The foregoing is merely the specific implementation case of patent of the present invention, but the technical characteristic of patent of the present invention is not limited to
This, any those skilled in the relevant art in the field of the invention, all cover in the special of the present invention by the change or modification made
Among sharp scope.
Sequence table
<110>Zhejiang Environmental Protection Research Academy
<120>A kind of microbial bacterial agent of organic waste fast degradation minimizing and its application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1389
<212> DNA
<213>Bacillus subtilis (Bacillus subtilis)
<400> 1
ggcttctaaa ggttacctca ccgacttcgg gtgttacaaa ctctcgtggt gtgacgggcg 60
gtgtgtacaa ggcccgggaa cgtattcacc gcggcatgct gatccgcgat tactagcgat 120
tccagctacg cagtcgagtt gcagactgcg atccgaactg agaacagatt tgtgggattg 180
gcttaacctc gcggtttcgc tgccctttgt tctgtccatt gtagcacgtg tgtagcccag 240
gtcataaggg gcagtgattt gacgtcatcc ccaccttcct ccggtttgtc accggcagtc 300
accttagagt gcccaactga atgctggcaa ctaagatcaa gggttgcgct cgttgcggga 360
cttaacccaa catctcacgc agagctgacg acaaccatgc accacctgtc actctgcccc 420
cgaaggggac gtcctatctc taggattgtc agaggatgtc aagacctggt aaggttcttc 480
gcgttgcttc gaattaaacc acatgtcccc gcttgtgcgg gcccccgtca attcctttga 540
gtttcagtct tgcgaccgta ctccccaggc ggagtgctta atgcgttagc tgcagcacta 600
aggggcggaa accccctaac acttagcact ctcgttacgg cgtggactac cagggtatct 660
aatcctgttc gctccccacg ctttcgctcc tcagcgtcag ttacagacca gagagtcgcc 720
ttcgccactg gtgttcctcc acatctctac gcatttcccg ctcacgtgga attccactct 780
cctcttctgc actcaagttc cccagtttcc aatgaccctc cccggttgag ccgggggctt 840
tcacatcaga cttaagaaac cgcctgcgag ccctttacgc ccataattcg gacaacgctt 900
gccacctacg tattaccgcg gctgctggca cgtagttagc cgtggctttc tggttaggta 960
ccgtcaaggt gccgccctat ttgaacggca cttgttcttc cctaacaacg agctttcgat 1020
ccgaaaacct tcatcactca cgcggcgttg ctccgtcaga ctttcgtcca ttgcggaaga 1080
ttccctactg ctgcctcccg taggagtctg ggccgtgtct cagtcccagt gtggccgatc 1140
accctctcag gtcggctacg catcgtcgcc ttggtgagcc gttacctcac caactagcta 1200
atgcgccggg tccatctgta agtggtagcc gaagccacct tttatgtctg aaccatgcgg 1260
ttcaaacaac catccggtat tagccccggt ttcccggagt tatcccagtc ttacaggcag 1320
gttacccacg tgtatcaccc gtccgccgct aacatcaggg agcaagctcc catctgtccg 1380
ctcgactgc 1389
<210> 2
<211> 1387
<212> DNA
<213>Bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
<400> 2
gcggctggct cctaaaggtt acctcaccga cttcgggtgt tacaaactct cgtggtgtga 60
cgggcggtgt gtacaaggcc cgggaacgta ttcaccgcgg catgctgatc cgcgattact 120
agcgattagc ttcacgcagt cgagttgcag actgcgatcc gaactgagaa cagatttgtg 180
ggattggctt aacctcgcgg tttcgctgcc ctttgttctg tccattgtag cacgtgtgta 240
gcccaggtca taaggcatga tgatttgacg tcatccccac cttcctccgg tttgtcaccg 300
gcagtcacct tagagtgccc aactgaatgc tggcaactaa gatcaagggt tgcgctcgtt 360
gcgggactta acccaacatt ccgacacgag ctgacgacaa ccatgcacca cctgtcactc 420
tgcccccgaa ggggacgtcc tatctctagg attgtcagag gatgtcaaga cctggtaagg 480
ttcttcgcgt tgcttcgaat taaacacagc tccaccgctt gtgcgggccc ccgtcaattc 540
ctttgagttt cagtcttgcg accgtactcc ccaggcggag tgcttaatgc gttagctgca 600
gcactaaggg gcggaaaccc cctaacactt acactatcgt ttacggcgtg gactaccagg 660
gtatctaatc ctgttcgctc cccacgcttt cgctcctcag cgtcagttac agaccagaga 720
gtcgccttcg ccactggtgt tcctcccaca tctctaccat ttaccgctac acgtggaatt 780
ccactctcct cttctgcact caagttcccc cagtttccaa tgaccctccc cggttgagcc 840
gggggctttc acatcagact taagaaaccg cctgcgagcc cttacgccca taattccgga 900
caacgcttgc cacctacgta ttaccgcggc tgctggcacg tagttagccg tggctttctg 960
gttaggtacc gtcaaggtgc cgccctattt gaacggcact tgttcttcct aacaacgagc 1020
tttacgatcc gaaaaccttc atcactcacg cggcgttgct ccgtcagact ttcgtccatt 1080
gcggaagatt ccctactgct gcctcccgta ggagtctggg ccgtgtctca gtcccgtgtg 1140
gccatcaccc tctcaggtcg gctacgcatc gtcgccttgg tgagccgtta cctcaccaac 1200
tagctaatgc gccgcgggtc catctgtaag tggtagccga agccaccttt tatgtctgaa 1260
catgcggttc aacaaccatc cggtattagc cccggtttcc cggagttatc ccagtcttac 1320
aggcaggtta cccacgtgtt actcacccgt ccgccgctaa catcagggag caagctccca 1380
tctgtcc 1387
Claims (10)
1. a kind of microbial bacterial agent of organic waste fast degradation minimizing, it is characterised in that by preserving number be CGMCC
The bacillus subtilis (Bacillus subtilis) of No.14052, the solution starch gemma that preserving number is CGMCC No.14053
Bacillus (Bacillus amyloliquefaciens), the bacillus subtilis (Bacillus that preserving number is CICC No.10210
Subtilis) it is mixed with preserving number for the cellulose-degrading bacteria (Bacillus sp) of CICC No.10829.
A kind of 2. method of organic waste fast degradation minimizing, it is characterised in that include the following steps：
(1) microbial bacterial agent as claimed in claim 1 is mixed with carrier, microbial bacterial agent is attached on carrier；By organic rubbish
Press filtration is handled after rubbish screening；
(2) organic waste after press filtration is handled is added in reaction warehouse, and the load of attached microbial microbial inoculum is added into reaction warehouse
Body, intermittent stirring reaction.
3. method according to claim 2, it is characterised in that the carrier is rice chaff.
4. method according to claim 2, it is characterised in that by the bulky grain organic waste screened out through powder in step (1)
After broken in input reaction warehouse.
5. method according to claim 2, it is characterised in that organic waste moisture control after press filtration processing 20~
40%.
6. method according to claim 2, it is characterised in that reaction condition is in reaction warehouse：Stir 5~10min, interval 10
~15min, mixing speed are 5~20rpm, and reaction temperature is 34~45 DEG C, and reactant pH is controlled 3.5~5.5.
7. method according to claim 2, it is characterised in that to every a batch of organic waste, interval is stirred in reaction warehouse
The time of reaction is 12~24h.
8. method according to claim 2, it is characterised in that the dosage of microbial bacterial agent is with microbial bacterial agent and reaction warehouse
The mass ratio of interior day output is 0.3~0.5% meter, is added once at interval of 10~15 days.
9. method according to claim 2, it is characterised in that the liquid produced after step (2) processing is after Anaerobic Treatment
It is discharged into municipal sewage pipe network.
10. method according to claim 2, it is characterised in that the stink produced in step (2) processing procedure is pumped into be equipped with and removes
Sewage network is discharged into after being purified in smelly dose of equipment.
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