CN109957527A - A kind of direct decomposing and fermenting crops straws produce special microorganism composite bacterial agent preparation and its application method of biogas - Google Patents
A kind of direct decomposing and fermenting crops straws produce special microorganism composite bacterial agent preparation and its application method of biogas Download PDFInfo
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Abstract
The invention discloses special microorganism composite bacterial agent preparation and its application methods that a kind of direct decomposing and fermenting crops straws produce biogas.This special microorganism composite bacterial agent is liquid or solid-state, by one plant or several plants in wax gemma bar, bacillus subtilis, bacillus licheniformis, bacillus megaterium, bacillusmusilaginosiengineering, Bacillus circulans, spherical bacillus, clostridium sporogene, acetone clostridium butyricum, Clostridium baratii, Clostridium beijerinckii, Clostridium thermocellum, Trichoderma viride, Filamentous fungi, aspergillus niger, after fermented culture, combination is compounded.Meanwhile using this special microorganism composite bacterial agent as inoculation microbial inoculum, provides and resolution process is carried out to agricultural crop straw, accelerate the application method of fermentation methane production；It is fast through microbial bacterial agent treated fermenting crops straws speed, produce that methane quantity is big, and processing cost is low, and without secondary pollution, can also ferment under conditions of without any fowl, animal manure just and generate biogas, the scope of application is very extensive.
Description
Technical field
The present invention relates to a kind of direct decomposing and fermenting crops straws produce biogas special microorganism composite bacterial agent preparation and
Its application method, the preparation for the microbial bacterial agent that especially cellulose family agricultural crop straw is decomposed；And it is dedicated multiple using this
Combined bacteria agent provides as inoculation microbial inoculum and carries out resolution process to agricultural crop straw, accelerate the application method of fermentation methane production.Simultaneously
It can be used for carrying out other organic solid wastes biological treatment production biogas.
Background technique
In order to improve Ecological Environment, the quality of life of peasant is improved, country widelys popularize the life of clean energy resource-biogas
It produces and utilizes.Currently, China rural area mainly produces biogas using poultry, poultry manure, according to statistics, due to each peasant household cultivation poultry, fowl number
Amount is uneven, larger in Various Seasonal peasant household cultivation quantity variance, therefore produces biogas, hair using agricultural crop straw as fermentation raw material
Residue, biogas slurry after ferment production biogas are good organic fertilizer again, and the method is worth promoting.But directly it is by agricultural crop straw
Fermenting raw materials produce biogas, can frequently result in produce biogas it is slow, gas production is few, factor of created gase is low, there is also fermentation materials meta-acid sometimes
Phenomena such as, it is subject to certain restrictions so promoting.
Currently, the preparation of complex microbial inoculum still is limited to the production of microbial-bacterial fertilizer etc., still without dedicated decomposition
Agricultural crop straw carries out the preparation of the complex microbial inoculum of the high-efficiency environment friendly of fermentation methane production.Currently, agricultural crop straw is handled
Method mainly has physical method, chemical method.Physical method mainly has mechanical mixing method, gas explosion method；Chemical method mainly uses alkali process, uses alkali
Measure larger, water consumption is also big；The above various technologies have that energy consumption is larger, and higher cost, rural area is poor for applicability and causes environment
The disadvantages of pollution.Therefore, the fermentation of bioanalysis resolution process is carried out to agricultural crop straw using special microorganism composite bacterial agent and produces natural pond
Gas will be well worth doing.
The biological process of biogas fermentation can be divided into three phases, i.e. hydrolysis stage, the sour rank of production by multi-cultur es synergy
Section produces the gas stage.It is produced in process of methane in fermenting crops straws, cellulose substances decomposition is crucial, the destruction of cellulose
Mean that complicated hydrolysis of organic matter is generated at corresponding monomer and incorporeity substance, this is most important physical behavior variation.
Therefore, it according to the characteristic of microbial strains and marsh gas fermentation processes principle, studies, to prepare dedicated high-efficiency environment friendly microorganism compound
It is inoculated with microbial inoculum, while providing and accelerating the processing side decomposed rich in cellulose agricultural crop straw using complex microbial inoculum
Method is conducive to the progress of marsh gas fermentation processes hydrolysis stage, and accelerates production acid phase and produce the implementation of gas stage, when shortening production biogas
Between, increase biogas output, is very necessary.
Summary of the invention
The purpose of the present invention is to propose to the special microorganism compound bacterias that a kind of direct decomposing and fermenting crops straws produce biogas
The preparation of agent, while providing and decomposing the application side that crop stalk carries out processing fermentation methane production using this complex microbial inoculum
Method；It effectively overcomes defect that is bad and influencing biogas output, yield of fermenting in the prior art, handles through microbial bacterial agent
Crop stalk fermenting speed afterwards is fastly and production methane quantity is big, makes initially to produce 50% or more gas time shortening, biogas output improves
30% or more；The agricultural crop straws such as all cornstalks, straw, straw can be made as the raw material of biogas fermentation, with existing other places
Reason method is compared, and processing cost is low, can be handled beside Farm House, field, methane-generating pit, and without secondary pollution；Straw
Methane-generating pit fermentation can be entered after stalk processing and generate biogas, biogas can be also generated under conditions of without any fowl, animal manure just, be applicable in model
It encloses very extensive.
The present invention is realized by following technical measures:
Bacterium that special microorganism composite bacterial agent of the present invention uses, fungi strain are by the whole in the 7-15 kind in following 15 kinds of bacterium
Or in which part forms.
Strain Chinese strain Classification system:
1 wax gemma bar Bacillus cereus
2 bacillus subtilis Bacillus subtilis
3 bacillus licheniformis Bacillus licheniformis
4 bacillus megaterium Bacillus megaterium
5 bacillusmusilaginosiengineering Bacillus mucilaginosus
6 Bacillus circulans Bacillus circulans
7 spherical bacillus Bacillus sphaericus
8 clostridium sporogene Clostridium sporogenes
9 acetone clostridium butyricum Clostridium acetobutylicum
10 Clostridium baratii Clostridium pasteurianum
11 Clostridium beijerinckii Clostridium beijerinckii
12 Clostridium thermocellum Clostridium thermocellum
13 Trichoderma viride Trichoderma viride
14 Filamentous fungi Trichoderma pseudokoningii
15 aspergillus niger Aspergillus niger
For convenience of description, above-mentioned 15 kinds of bacterium are indicated with its serial number in this application, and such as No. 1 bacterium is wax bacillus, and No. 2 bacterium are
For bacillus subtilis, and so on.Above-mentioned strain can voluntarily separate identification or directly be bought by culture presevation unit and be used.
The above bacterial strain common ground is that bacterium can generate gemma, and fungi is high activity medium temperature, mesophilic cellulose-degrading bacteria；The above bacterial strain warp
Screening, activation, rejuvenation, mutagenesis, cellulose degradation vigor improve 50% or more.Above 15 kinds of bacterium in the present invention will be complete
Portion or part are applied in combination, and are prepared as product.The present invention guarantees that product has good storage, application reliability.
Microbial bacterial agent the preparation method comprises the following steps: each bacterium is carried out solid or liquid fermentation and culture alone or in combination, cultivate
Base is general production gemma culture medium or is generally conducive to bacterial growth, produces gemma culture medium；Each fungi is consolidated alone or in combination
Body or liquid fermentation and culture, culture medium are general fungi culture medium.Now the preparation method of solid and liquid bacterial agent is described as follows:
One, the preparation method of solid and liquid bacterial agent
1, the preparation method of solid fungicide
Bacteria culture fermented and cultured generates bacterial spore
1-7 strain is respectively or after combination, using general production gemma culture medium and condition of culture, such as glucose broth
Deng, carry out fermented and cultured, with generate gemma peak conditions determine terminate fermentation (at this time spore concentration generally 1 × 109-4 ×
1010/ml)；8-12 strain produces gemma culture medium and Anaerobic culturel item respectively or after combination, using general anaerobic bacteria
Part, such as cooked meat medium carry out fermented and cultured, determine termination fermentation (spore concentration at this time to generate the peak conditions of gemma
Generally in 1 × 109-4 × 1010/ml).
Bacterial spore is collected after fermentation and prepares gemma dry powder
According to conventional general method, after above-mentioned strain fermentation, the collection of gemma spore, powder processed are carried out.Adsorbent is added to filter
The methods of collect bacterial spore spore, air-dried using 70 DEG C or less, spray drying, conidia powder, the appropriate suction of cooperation be made after dry
Attached dose, dilution filler at preparation, measure effective spore concentration.Adsorbent is diatomite, precipitated calcium carbonate, careless carbon, Power Plant Ash etc.
The public knows substance.
Fungi strain solid fermentation culture
One plant in 13-15 or several plants of medium temperatures, mesophilic cellulose degradation fungi, respectively or combination is inoculated in the dedicated training of solid state fermentation
It supports and is cultivated according to a conventional method in base, obtain solid state fermentation microbial inoculum (bacteria containing amount is > 109cfu/g)；The dedicated training of solid state fermentation
Base is supported to be compounded by sawdust, rice bran, wheat bran, inorganic salts.
Preparing microorganism microbial inoculum and packaging
According to bacterial spore concentration and fungi concentration requirement, product is prepared by 1: 1；It is lower than in the product moisture content for guaranteeing preparation
10%, when pollution microbes are less than 5%, product final packaging can be carried out.
2, the preparation method of liquid bacterial agent
Bacteria culture fermented and cultured generates bacterial spore
1-7 strain is respectively or after combination, using general production gemma culture medium and condition of culture, such as glucose broth
Deng, carry out fermented and cultured, with generate gemma peak conditions determine terminate fermentation (at this time spore concentration generally 1 × 109-4 ×
1010/ml)；8-12 strain produces gemma culture medium and Anaerobic culturel item respectively or after combination, using general anaerobic bacteria
Part, such as cooked meat medium carry out fermented and cultured, and to generate, an armful peak conditions for son are determining to terminate fermentation (spore concentration at this time
Generally in 1 × 109-4 × 1010/ml).
Fungi strain liquid fermentation and culture
One plant in 13-15 or several plants of medium temperatures, mesophilic cellulose degradation fungi, respectively or combination is inoculated in the dedicated training of liquid state fermentation
It supports in base, is cultivated according to a conventional method, obtain liquid state fermentation microbial inoculum (bacteria containing amount is > 109cfu/ml).Liquid state fermentation is dedicated
Culture medium adds water to prepare by soya-bean milk, glucose.
Preparing microorganism liquid bacterial agent product
According to each bacterial spore concentration and formulation concentrations requirement, protective agent and sterile water and fungi fermentation is added to train with above-mentioned spore liquid
Nutrient solution filling is configured to liquid product by 1: 1.
Two, the application method of resolution process fermentation methane production is carried out to agricultural crop straw using special microorganism composite bacterial agent
The present invention carries out resolution process fermentation methane production, stalk internal structure that treated to agricultural crop straw using composite bacteria agent
It changes, stalk limbers up, is loose, so stalk can be made conducive to anaerobe degradation as the raw material for producing biogas
It utilizes, makes initially to produce 50% or more gas time shortening, biogas output improves 30% or more.Because the bacterium of composite bacteria agent used in it,
In fungi, containing thermophilic, medium temperature cellulose-degrading bacteria, therefore the synergistic effect of each bacterium is utilized, it can be in the item without any fowl, animal manure just
Biogas all is generated using stalk fermentation under part.
Decomposing process is carried out to stalk using special microorganism composite bacterial agent are as follows: first break crop stalk raw material
It is broken, add water to soak, appropriate urea water then is added to the straw after wetting or the nitrogen fertilizer in agriculture such as carbon ammonia or tapioca starch, waste matter slag are made
It is supplemented for nitrogen source, while special microorganism composite bacterial agent is added, mixed thoroughly, carried out the modes such as stack retting and handle, stack retting temperature is not less than
50 DEG C maintain 3 days or more, then will through special microorganism composite bacterial agent, treated that crop stalk enters methane-generating pit, it is appropriate to be added
Biogas fermentation inoculum and special-purpose catalyst, closed anaerobic fermentation produce biogas.The total solid concentration of fermentation materials is adjusted to 15-
50%, carry out anaerobism producing methane by dry-type fermentation；The total solid concentration of fermentation materials is adjusted to 15% hereinafter, carrying out anaerobism wet type
Fermentation methane production.
Specific embodiment
Embodiment one, the preparation of special microorganism solid fungicide
Bacteria culture fermented and cultured
1-7 strain is carried out to rejuvenation, ultraviolet mutagenesis (the above technology is known in those skilled in the art) according to a conventional method respectively,
Individually use following condition fermented and cultureds: culture medium wins biotechnology for bacteriotrophy broth bouillon, purchased from Qingdao sea
Company, by specification are configured to 10L, PH7.2, are packed into 18 liters of automatic fermenters；Condition of culture are as follows: 30 DEG C of cultivation temperature；It stirs
Mix 200 revs/min of speed；Filtrated air ventilatory capacity 1: 1；Incubation time 24 hours；Spore concentration > 1 × 109 when fermentation ends
A/ml.
8-12 strain individually uses following condition fermented and cultureds: culture medium is bacterium anaerobism cooked meat medium, purchase
From Qingdao Hai Bo biotech company, by specification is configured to 10L, PH7.2, is packed into 18 liters of automatic fermenters；Condition of culture
Are as follows: 30 DEG C of cultivation temperature；200 revs/min of mixing speed: sterile nitrogen ventilatory capacity 0.7: 1；Incubation time 20 hours；Culture knot
Spore concentration is 2 × 109/ml of > when beam.
Gemma dry powder is collected and prepared to bacterial spore after fermentation
After above-mentioned strain fermentation plus diatomite uses centrifugal filtration, collects gemma thallus；300 are made after 60 DEG C of heated-air dryings
Mesh, tune thallus spore concentration are 1 × 1010/g, prepare bacterium solid-state microorganism microbial inoculum by equal weight proportions.Fungi strain fermentation
Culture
13-15 strain individually uses following condition fermented and cultureds: by No. 13-15 Trichoderma viride, Filamentous fungi, black song
Mould each one plant carries out rejuvenation, ultraviolet mutagenesis (the above technology is known in those skilled in the art) according to a conventional method, individually adopts
With following condition fermented and cultureds: (inoculum concentration 10%) is individually inoculated in solid state fermentation special culture media, at 32 DEG C
Culture 48 hours, obtains solid-state version fungi fermentation microbial inoculum.Solid state fermentation special culture media specific formula are as follows: sawdust 4kg, rice bran
1kg, wheat bran 1kg, (NH4) 2SO45g, KH2PO42.5g add water to water content up to 50%, PH 7.0.Above-mentioned fungi solid-state microbial inoculum
Soft for yellow, bacteria containing amount > bacteria containing amount is > 109cfu/g.
Complex microbial inoculum compounding and packaging
Above-mentioned bacterium, fungi solid-state microorganism microbial inoculum are compounded in 1: 1 ratio, and above-mentioned water content is lower than 10% after detection, are polluted miscellaneous
When bacterium is less than 5%, product final packaging is carried out；It is packaged as 500g/ bags.
The preparation method of embodiment two, microbial liquid microbial inoculum
Bacteria culture fermented and cultured
1-7 strain is carried out to rejuvenation, ultraviolet mutagenesis (the above technology is known in those skilled in the art) according to a conventional method respectively,
Individually use following condition fermented and cultureds: culture medium wins biotechnology for bacteriotrophy broth bouillon, purchased from Qingdao sea
Company, by specification are configured to 10L, PH7.2, are packed into 16 liters of automatic fermenters；Condition of culture are as follows: 30 DEG C of cultivation temperature；It stirs
Mix 200 revs/min of speed；Filtrated air ventilatory capacity 1: 1；Incubation time was at 24 hours；Spore concentration > 1 when fermentation ends ×
109/ml.
8-12 strain individually uses following fermentation culture conditions: culture medium is purchased from for bacterium anaerobism cooked meat medium
Qingdao Hai Bo biotech company, by specification are configured to 10L, PH7.2, are packed into 16 liters of automatic fermenters；Condition of culture are as follows:
30 DEG C of cultivation temperature；200 revs/min of mixing speed: sterile nitrogen ventilatory capacity 0.7: 1；Incubation time 20 hours；At the end of culture
Spore concentration is 2 × 109/ml of >.
Fungi strain fermented and cultured
13-15 strain individually uses following condition fermented and cultureds: by No. 13-15 Trichoderma viride, Filamentous fungi, black song
Mould each one plant carries out rejuvenation, ultraviolet mutagenesis (the above technology is known in those skilled in the art) according to a conventional method, individually adopts
With following condition fermented and cultureds: (inoculum concentration 10%) is individually inoculated in liquid fermentation special culture media 10L, at 32 DEG C
Lower culture 64 hours；Liquid state fermentation special culture media formula: the soya-bean milk that mass/volume percent concentration is 80%, glucose
20g adds water to 10L, pH 7.0.
Microbial bacterial agent compounding and packaging
According to each spore concentration and formulation concentrations requirement, protective agent and sterile water and fungi strain fermentation is added to train with above-mentioned gemma liquid
Nutrient solution body filling is configured to liquid product by 1: 1；It is packaged as 500ml/ bottles.
Claims (6)
1. a kind of direct decomposing and fermenting crops straws produce special microorganism composite bacterial agent preparation and its application method of biogas,
It is characterized in that: the special microorganism composite bacterial agent of direct decomposing straw fermentation methane production, by being capable of degraded cellulose, hemicellulose
The bacterium for generating gemma of element, lignin, including wax gemma bar, bacillus subtilis, bacillus licheniformis, huge gemma bar
Bacterium, bacillusmusilaginosiengineering, Bacillus circulans, spherical bacillus, clostridium sporogene, acetone clostridium butyricum, Clostridium baratii,
Clostridium beijerinckii, Clostridium thermocellum, in one plant or several plants；By can degraded cellulose, hemicellulose, lignin medium temperature, mesophilic
Fungi, including one plant or several plants in Trichoderma viride, Filamentous fungi, aspergillus niger, after fermented culture, combination is compounded；Together
When provide using this special microorganism composite bacterial agent to agricultural crop straw progress resolution process, accelerate the application side of fermentation methane production
Method.
2. direct decomposing and fermenting crops straws according to claim 1 produce the special microorganism composite bacterial agent of biogas,
It is characterized in that the special microorganism composite bacterial agent is liquid or solid-state.
3. special microorganism composite bacterial agent according to claim 1 or 2, it is characterised in that the liquid microbial inoculum preparation method
Are as follows: one plant in bacterium or several plants of bacterium are inoculated in aerobic in liquid state fermentation special culture media or Anaerobic culturel alone or in combination, good
Oxygen bacterium cultivates logical filtrated air, and anaerobic bacteria culture leads to sterile nitrogen, obtains liquid state fermentation microbial inoculum；In one plant or several plants in fungi
Warm, mesophilic cellulose-degrading bacteria is inoculated in aerobic culture in liquid state fermentation special culture media alone or in combination, obtains liquid state fermentation
Microbial inoculum；By above-mentioned microbial inoculum by 1: 1 mixing compounding packaging.
4. special microorganism composite bacterial agent according to claim 1 or 2, it is characterised in that the solid-state microbial inoculum preparation method
Are as follows: one plant in bacterium or several plants of bacterium are inoculated in alone or in combination in liquid state fermentation special culture media, after culture plus adsorbent warp
Centrifugation or plate-frame filtering collect gemma and thallus, using 70 DEG C it is below air-dry or spray drying, cooperate appropriate adsorbent and dilute
It releases filler and pulvis is made；One plant in fungi or several plants of medium temperatures, mesophilic cellulose-degrading bacteria are inoculated in solid-state hair alone or in combination
It is cultivated in ferment special culture media, obtains solid-state version fermenting agent；By above-mentioned microbial inoculum by 1: 1 mixing compounding packaging.
5. according to claim 1 decompose agricultural crop straw quickening fermentation methane production using special microorganism composite bacterial agent
Application method, it is characterised in that decompose the application method of straw fermentation methane production are as follows: first crop stalk raw material is crushed, adds water
Wetting is added special microorganism microbial inoculum, adds urea water or carbon ammonia or other nitrogen fertilizer in agriculture in right amount, mix thoroughly, carries out the modes such as stack retting
Processing, stack retting temperature maintain 3 days or more not less than 50 DEG C, then will treated that crop stalk enters through special microorganism microbial inoculum
Methane-generating pit, appropriate that biogas fermentation inoculum or special-purpose catalyst is added, closed anaerobic fermentation produces biogas.
6. decomposing the application method of straw fermentation methane production according to claim 1 or 5, it is characterised in that the warp is specially
With microbial bacterial agent, treated that crop stalk enters after methane tank straw enters methane-generating pit, can be dense by the total solid of fermentation materials
Degree is adjusted to 15-50%, carries out dry-type anaerobic fermentation and produces biogas；The total solid concentration of fermentation materials can also be adjusted to 15%
Hereinafter, carrying out wet type producing methane through anaerobic fermentation.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184228A (en) * | 2019-07-10 | 2019-08-30 | 山东省农业科学院农业资源与环境研究所 | A kind of method of corn stover anaerobic dry fermentation microbial inoculum and biogas slurry reflux volume increase methane |
| CN111004748A (en) * | 2019-12-26 | 2020-04-14 | 青岛尚德生物技术有限公司 | Microbial strengthening microbial inoculum capable of remarkably promoting fermentation effect before household garbage incineration and application thereof |
| CN112624866A (en) * | 2021-01-01 | 2021-04-09 | 浙江大学 | Organic slow release fertilizer using livestock and poultry manure and straw as raw materials |
| CN113088462A (en) * | 2021-03-12 | 2021-07-09 | 浙江大学 | Composite leaven for organic material compost |
-
2017
- 2017-12-26 CN CN201711428712.3A patent/CN109957527A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184228A (en) * | 2019-07-10 | 2019-08-30 | 山东省农业科学院农业资源与环境研究所 | A kind of method of corn stover anaerobic dry fermentation microbial inoculum and biogas slurry reflux volume increase methane |
| CN110184228B (en) * | 2019-07-10 | 2024-04-02 | 山东省农业科学院农业资源与环境研究所 | Corn straw anaerobic dry fermentation microbial inoculum and biogas slurry reflux methane production method |
| CN111004748A (en) * | 2019-12-26 | 2020-04-14 | 青岛尚德生物技术有限公司 | Microbial strengthening microbial inoculum capable of remarkably promoting fermentation effect before household garbage incineration and application thereof |
| CN112624866A (en) * | 2021-01-01 | 2021-04-09 | 浙江大学 | Organic slow release fertilizer using livestock and poultry manure and straw as raw materials |
| CN113088462A (en) * | 2021-03-12 | 2021-07-09 | 浙江大学 | Composite leaven for organic material compost |
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