CN105969835B - Method for preparing methane by straw and excrement combined fermentation - Google Patents

Method for preparing methane by straw and excrement combined fermentation Download PDF

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CN105969835B
CN105969835B CN201610460822.7A CN201610460822A CN105969835B CN 105969835 B CN105969835 B CN 105969835B CN 201610460822 A CN201610460822 A CN 201610460822A CN 105969835 B CN105969835 B CN 105969835B
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fermentation
microbial inoculum
pseudomonas
straw
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CN105969835A (en
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崔宗均
袁旭峰
朱万斌
朱金波
师力民
姚建明
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Middle Green (beijing) Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/02—Biological treatment
    • C02F11/04—Anaerobic treatment; Production of methane by such processes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
    • C12P5/023—Methane
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
    • 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
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

The invention discloses a method for preparing methane by combined fermentation of straw and excrement. The invention provides a methane fermentation method, which comprises the following steps: fermenting the raw materials by using a microbial inoculum to realize methane fermentation; the raw materials consist of straws, animal wastes and water; the mass ratio of C atoms in the straw to N atoms in the animal manure is 20-35: 1. experiments prove that the method for preparing methane by combined fermentation of crop straws and animal wastes overcomes the problems that the existing biogas engineering taking straws as raw materials is unreasonable in nutrient proportion, lacks of constructing high-efficiency decomposition microbial communities, is unreasonable in straw decomposition process and the like. The method realizes high-efficiency decomposition and acidification, and then transfers the decomposition and acidification into anaerobic fermentation to realize high-efficiency biogas production.

Description

Method for preparing methane by straw and excrement combined fermentation
Technical Field
The invention relates to the technical field of biology, in particular to a method for preparing methane by straw and excrement combined fermentation.
Background
Methane fermentation is a process in which organic substances are decomposed into small molecular organic acids under anaerobic conditions and then methane is generated. Mankind controls and projects the process to form 'methane fermentation engineering', or 'methane fermentation engineering'. The methane fermentation engineering can obtain clean energy, namely methane, while treating the environmental pollution caused by organic wastes. The main components of the biogas are methane, CO2 and hydrogen sulfide. The biogas can be converted into 'biogas' with methane content reaching a certain standard through the purification process of removing CO2 and hydrogen sulfide.
The methane fermentation microorganisms mainly utilize water-soluble organic matters with smaller molecules. Therefore, the traditional methane fermentation raw materials mainly comprise various waste water containing organic matters, animal manure, organic matter processing waste residues and the like. With the maturity of the purification technology of 'biogas', the aim of methane fermentation engineering is mainly transferred from environmental management to energy production. Accordingly, the traditional methane fermentation raw materials can not meet the requirements of engineering development. Therefore, the raw materials containing cellulose, such as straws, pasture, bagasse and the like attract attention of people, and various biogas fermentation technologies utilizing the straws appear. However, the methane-fermenting microorganisms cannot directly decompose and utilize the cellulose-containing raw material, and the conversion rate of the cellulose-containing raw material and the equipment volume methane-producing efficiency are extremely unstable. The main problems are as follows:
① the nutrient of the straw is monotonous, the nutrient proportion is unreasonable, the microorganism decomposes and utilizes the crop straw, the reasonable nutrient proportion is needed, especially the proportion of C and N is reasonable in 20-35, and certain phosphorus element and microelement are needed.
② the method is lack of microorganism group for decomposing straw efficiently, which not only needs reasonable nutrient proportion, but also needs to construct efficient microorganism group for decomposing straw and meet the growth condition of the microorganism group in the process of straw decomposition.
③ the decomposition process of straw is not reasonable, to achieve the high-efficiency decomposition of straw, besides the reasonable nutrient proportion and high-efficiency microbiota, it also needs to satisfy the environment condition and fermentation process suitable for the growth of high-efficiency decomposition microbiota.
Disclosure of Invention
It is an object of the present invention to provide a method for methane fermentation.
The method provided by the invention comprises the following steps: fermenting the raw materials by using a microbial inoculum to realize methane fermentation;
the raw materials consist of straws, animal wastes and water;
the mass ratio of C element to N element of the raw material is 20-35: 1;
the active ingredients of the microbial inoculum are Clostridium xylanolyticum (Clostridium xylanolyticum), cellulolytic Pseudomonas (Xylanimonas cellulilytica), Pseudomonas (Pseudomonas sp.) and Bacillus subtilis (Bacillus subtilis).
In the method, the mass ratio of the C element to the N element of the raw material is 20: 1. 25:1 or 30: 1.
in the above method, the mass ratio of the straw, the animal waste and the water is 1: 0.5-1: 8-8.5.
In the above method, the mass ratio of the straw, the animal waste and the water is 1: 1: 8 or 1: 0.5: 8.5.
in the method, the straw is corn straw, wheat straw or rice straw.
In the above method, the animal waste is cow waste, pig waste or chicken waste.
In the method, the straws are straw particles with the length less than 1 cm.
In the method, the mass ratio of the straw to the animal waste to the water to the microbial inoculum is 1: 0.5-1: 8-8.5: 0.001.
the mass ratio of the straw to the animal waste to the water to the microbial inoculum is 1: 1: 8: 0.001 or 1: 0.5: 8.5: 0.001.
the microbial inoculum is prepared by uniformly mixing a Clostridium xylanolyticum (Clostridium xylanolyticum) culture solution, a cellulolytic xylan unicellular bacteria (xylomonas) culture solution, a Pseudomonas sp (Pseudomonas sp) culture solution and a Bacillus subtilis culture solution to obtain a mixed bacterial liquid, and adsorbing the mixed bacterial liquid on a carrier to obtain the microbial inoculum.
In the microbial inoculum, the ratio of the Clostridium xylanisolvens (Clostridium xylanolyticum), the cellulomonas xylanolyticus (xylomonas cellulolytica), the Pseudomonas (Pseudomonas sp.), the Bacillus subtilis (Bacillus subtilis) and the carrier is (25-35) cfu: (25-30) cfu (20-30) cfu (15-20): 1X 10-4-1×10-3g；
Or, the ratio of the Clostridium xylanisolvens (Clostridium xylanolyticum), the cellulomonas xylanolyticus (xylolysimachia), the Pseudomonas (Pseudomonas sp.), the Bacillus subtilis (Bacillus subtilis) and the carrier is (25-35) cfu: (25-30) cfu (20-30) cfu:20cfu: 5X 10-4g；
Or the content of the Clostridium xylanilyticum (Clostridium xylolyticum) in the microbial inoculum is 50000-70000 cfu/g;
the content of the cellulolytic xylan monad (Xylanimonas cellulolysitica) in the microbial inoculum is 50000-60000 cfu/g;
the content of the Pseudomonas (Pseudomonas sp.) in the microbial inoculum is 40000-;
the content of the Bacillus subtilis in the microbial inoculum is 40000 cfu/g.
The carrier is crushed straw obtained by crushing dry rice straws and then sieving the crushed straw by a sieve of 0.4 mm.
In the microbial inoculum, the mass ratio of the mixed bacterial liquid to the carrier is 1: 10;
the colony forming unit ratio of each bacterium in the mixed bacterium liquid is (25-35): (25-30):(20-30):(15-20).
Or the colony forming unit ratio of each bacterium in the mixed bacterium liquid is (25-35): (25-30):(20-30):20.
In the method, the straws are corn straws, wheat straws or rice straws;
and/or the straw is straw particles smaller than 1 cm.
In the above method, the animal waste is cow waste, pig waste or chicken waste.
In the above method, the fermentation comprises the following steps:
1) uniformly mixing the microbial inoculum and the raw materials, stirring and fermenting to obtain a decomposition fermentation system;
the stirring fermentation conditions are that the stirring speed is 50 r/min, stirring is carried out for 10 minutes in each hour, then the stirring is stopped for 50 minutes, the stirring fermentation is carried out for 36 hours, and the temperature is 30 ℃;
2) fermenting the decomposition fermentation system in an anaerobic fermentation container to realize methane fermentation;
the anaerobic fermentation condition is 35 ℃ and 30 days.
In the above method, the Clostridium xylanolyticum (Clostridium xylanolyticum) is Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No. 6555; https:// www.dsmz.de).
The cellulolytic xylanase monad (Xylanimonas cellulolytica) is cellulolytic xylanase monad (Xylanimonas cellulolytica) DSM No. 15894; https:// www.dsmz.de).
The Pseudomonas (Pseudomonas sp.) is Pseudomonas ACCCC No. 11691.
The Bacillus subtilis (Bacillus subtilis) is Bacillus subtilis ACCC No. 03189.
Another object of the present invention is to provide a microbial inoculum for methane fermentation.
The microbial inoculum for methane fermentation provided by the invention is the microbial inoculum in the method.
The method of the invention has the following characteristics:
①, optimizing the mixture ratio of straw and animal waste:
the straw comprises corn straw, wheat straw and rice straw; the manure comprises cow manure, pig manure and chicken manure. The straws are crushed into particles with the diameter smaller than 1cm, the excrement is screened to remove large particles with the diameter larger than 1cm, and the sand particles are removed through water soaking and precipitation. When the straw and the excrement are proportioned, the straw, the excrement and water are uniformly mixed, so that the proportion of dry matters of the raw materials to the water is equal to 1/10, and the C/N of the raw materials is within the range of 20-35.
② construction of efficient decomposition microbial population:
in a cement pool with diameter of 4.2m, depth of 4m and central vertical paddle stirrer, adding straw and excrement to make straw decomposition system according to the above-mentioned mixing ratio, and utilizing the patent "a straw in-situ decomposition microbial inoculum and its application" (patent No. CN 103642721A), the inoculation ratio is 1m3Adding 100g of solid microbial inoculum into a decomposition system, uniformly scattering the microbial inoculum, uniformly stirring, and maintaining the temperature of 30 ℃ to construct a high-efficiency decomposition microbial community taking clostridium as a main body.
③ two-step fermentation process of first facultative anaerobic decomposition and then methane fermentation
Controlling the temperature of the decomposition fermentation system at 30 ℃, and controlling the stirring intensity at: stirring at 50 rpm for 10 min per hour, stopping stirring for 50 min to provide facultative anaerobic condition, pumping the decomposed fermented liquid into anaerobic fermenting tank with screw pump after 36 hr, maintaining the temperature in the fermenting tank at 35 deg.c for 30 days, and fermenting in a full mixed fermentation (CSTR) process.
Experiments prove that the method for preparing methane by combined fermentation of crop straws and animal wastes overcomes the problems that the existing biogas engineering taking straws as raw materials is unreasonable in nutrient proportion, lacks of constructing high-efficiency decomposition microbial communities, is unreasonable in straw decomposition process and the like. The method comprises the steps of proportioning straws and animal wastes according to an optimal proportion, utilizing high-efficiency microbial community decomposition, and adopting a two-step fermentation process, namely, firstly creating a facultative anaerobic condition suitable for microbial community decomposition in a fermentation system for decomposition and fermentation to realize high-efficiency decomposition and acidification, and then transferring the fermentation system into anaerobic fermentation to realize high-efficiency biogas production.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The preparation of the microbial inoculum in the following examples is disclosed in patent publication No. CN 103642721A, and the active ingredients are Clostridium xylanolyticum (Clostridium xylanolyticum) No. DSM No.6555(https:// www.dsmz.de), cellulolytic xylanolyticum (Xylanimmonas cellulolytica) No. DSM No.15894(https:// www.dsmz.de), Pseudomonas (Pseudomonas sp.) No. ACCC No.11691 (China agricultural microbial cultures Collection center (ACCC)), and Bacillus subtilis (ACCC No. 03189).
The microbial inoculum is prepared by uniformly mixing a Clostridium xylanolyticum (Clostridium xylanolyticum) culture solution, a cellulolytic xylan monad (Xylanimonas cellulolytica), a Pseudomonas sp (Pseudomonas sp) culture solution and a Bacillus subtilis (Bacillus subtilis) culture solution to obtain a mixed bacterial solution, and adsorbing the mixed bacterial solution onto a carrier to obtain the microbial inoculum.
Example 1 Combined methane fermentation of corn stover and cow dung
1. Microbial inoculum for quickly decomposing organic matters
The organic matter fast decomposing microbial inoculum is prepared by the following method:
respectively culturing Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, Clostridium cellulolyticum (Xylanimomonas cellulolytica) DSM No.15894, Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 and Bacillus subtilis (Bacillus subtilis) ACCC No.03189 to obtain a Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555 culture solution, a Clostridium cellulolyticum (Xylanimidolytica) DSM No.15894 culture solution, a Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 culture solution and a Bacillus subtilis (Bacillus subtilis) ACCC 03189 culture solution.
Mixing culture solution of Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, culture solution of cellulolytic xylanase (Xanthomonas cellulolytica) DSM No.15894, culture solution of Pseudomonas sp (Pseudomonas sp.) ACCC No.11691, and culture solution of Bacillus subtilis ACCC No.03189, and obtaining mixed bacteria solution.
Crushed straw obtained by crushing dry rice straws and passing the crushed straw through a 0.4mm sieve is used as a carrier.
And then spraying a mixed bacterial liquid accounting for 10 percent of the mass of the chopped grass while stirring in 100Kg of chopped grass, and adsorbing the mixed bacterial liquid on a carrier to obtain the organic matter fast-decomposing microbial inoculum.
In the microbial inoculum, the mixture ratio of Clostridium xylanilyticum (Clostridium xylanilyticum), cellulolytic Pseudomonas (Xylanimonas cellulolytica), Pseudomonas (Pseudomonas sp.), Bacillus subtilis (Bacillus subtilis) and a carrier is 25cfu: 25cfu:20cfu: 5X 10-4g；
Wherein the content of Clostridium xylanisolvens (Clostridium xylanolyticum) in the microbial inoculum is 50000 cfu/g;
the content of cellulolytic xylan monad (Xylanimonas cellulosilytica) in the microbial inoculum is 50000 cfu/g;
the content of Pseudomonas (Pseudomonas sp.) in the microbial inoculum is 40000cfu/g;
the content of the Bacillus subtilis in the microbial inoculum is 40000 cfu/g.
2. Combined methane fermentation of corn straw and cow dung
The implementation place is as follows: \28095TheChina agriculture university scientific and technological park industry biogas engineering platform of Zhou city of 60m3A full mixing mode;
the components required for fermentation are as follows: corn straw (water content 15%), cow dung (water content 85%), and the organic matter fast decomposing microbial inoculum obtained in the previous step;
the implementation method comprises the following steps: effective volume of decomposition fermentation tank is 50m3The effective volume of the full-mixing anaerobic fermentation tank is 60m3。
The raw material ratio is as follows: uniformly mixing 5 tons of corn straws, 5 tons of cow dung and 40 tons of water to obtain a raw material; the ratio of C to N of the raw materials is 25: 1; adding 5kg of the obtained organic matter fast decomposing microbial inoculum into 50 tons of raw materials;
crushing the straws into particles with the diameter less than 1cm, removing large particles with the diameter more than 1cm from excrement by a sieve, and adding the corn straws, the cow dung, the water and the microbial inoculum into an effective volume of 50m according to the proportion3Stirring the mixture in the decomposition fermentation tank for 30 minutes at a rotating speed of 50 revolutions per minute (rotating radius of 2m), uniformly mixing, controlling the temperature at 30 ℃, and controlling the stirring intensity at: the rotation speed is 50 r/min, stirring is carried out for 10 min in each hour, stopping for 50 min to provide facultative anaerobic condition, and after running for 36 hours, 40% dry matter of the decomposing and fermenting system becomes water soluble, and 60% dry matter remains in suspension state to obtain decomposing and fermenting mixed liquor.
Pumping the decomposition fermentation mixed solution into a screw pump of 60m3In an anaerobic fermentation tank, the temperature of the fermentation tank is kept at 35 ℃, the liquid retention time is 30 days, and methane fermentation is carried out by a full mixed fermentation (CSTR) fermentation process to obtain a fermentation product.
The residual dry matter in 1L of the fermentation product is detected by a dry matter loss method (in detail, see the literature: Yuan Peak, Gao Rui Fang, Li culture and the like. the improvement of the methane production efficiency of the anaerobic fermentation of the corn straws by the pretreatment of a compound strain MC1, 2011, 9 (27): 266-.
As a result, the conversion rate of the dry matters of the straws and the cow dung is more than 50 percent, and the daily methane yield reaches 90m3The above.
Example 2 Combined methane fermentation of wheat straw and pig manure
1. Microbial inoculum for quickly decomposing organic matters
The organic matter fast decomposing microbial inoculum is prepared by the following method:
respectively culturing Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, Clostridium cellulolyticum (Xylanimomonas cellulolytica) DSM No.15894, Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 and Bacillus subtilis (Bacillus subtilis) ACCC No.03189 to obtain a Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555 culture solution, a Clostridium cellulolyticum (Xylanimidolytica) DSM No.15894 culture solution, a Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 culture solution and a Bacillus subtilis (Bacillus subtilis) ACCC 03189 culture solution.
Mixing culture solution of Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, culture solution of cellulolytic xylanase (Xanthomonas cellulolytica) DSM No.15894, culture solution of Pseudomonas sp (Pseudomonas sp.) ACCC No.11691, and culture solution of Bacillus subtilis ACCC No.03189, and obtaining mixed bacteria solution.
Crushed straw obtained by crushing dry rice straws and passing the crushed straw through a 0.4mm sieve is used as a carrier.
And then spraying a mixed bacterial liquid accounting for 10 percent of the mass of the chopped grass while stirring in 100Kg of chopped grass, and adsorbing the mixed bacterial liquid on a carrier to obtain the organic matter fast-decomposing microbial inoculum.
In the microbial inoculum, the ratio of Clostridium xylanilyticum (Clostridium xylanilyticum), cellulolytic Pseudomonas (Xylanimonas cellulolytica), Pseudomonas (Pseudomonas sp.), Bacillus subtilis (Bacillus subtilis) and carrier is 35 cfu:30 cfu:30cfu:20 cfu: 5X 10-4g；
Wherein the content of Clostridium xylanisolvens (Clostridium xylanolyticum) in the microbial inoculum is 70000 cfu/g;
the content of cellulolytic xylan monad (Xylanimonas cellulosilytica) in the microbial inoculum is 60000 cfu/g;
the content of Pseudomonas (Pseudomonas sp.) in the microbial inoculum is 60000 cfu/g;
the content of the Bacillus subtilis in the microbial inoculum is 40000 cfu/g.
2. Combined methane fermentation of wheat straw and pig manure
The implementation place is as follows: 28095Zhou Chinese agricultureScientific and technological garden industry biogas engineering platform 60m3Full mixing type
The components required for fermentation are as follows: wheat straw (water content 12%), pig manure (water content 88%), and the organic matter fast decomposing microbial inoculum prepared in the step 1;
the implementation method comprises the following steps: effective volume of decomposition fermentation tank is 50m3The effective volume of the full-mixing anaerobic fermentation tank is 60m3。
The raw material ratio is as follows: uniformly mixing 5 tons of wheat straws, 5 tons of pig manure and 40 tons of water to obtain a raw material; the ratio of C to N of the raw materials is 30: 1;
adding 5kg of the organic matter fast decomposing microbial inoculum prepared in the step 1 into 50 tons of raw materials;
crushing the straws into particles with the diameter less than 1cm, removing large particles with the diameter more than 1cm from excrement by a sieve, and adding the corn straws, the cow dung, the water and the microbial inoculum into an effective volume of 50m according to the proportion3The decomposition fermentation tank is stirred for 30 minutes at the rotating speed of 50 r/min and uniformly mixed, the temperature is controlled at 30 ℃, and the stirring intensity is controlled as follows: the rotation speed is 50 r/min, stirring is carried out for 10 min in each hour, stopping for 50 min to provide facultative anaerobic condition, and after running for 36 hours, 40% dry matter of the decomposing and fermenting system becomes water soluble, and 60% dry matter remains in suspension state to obtain decomposing and fermenting mixed liquor.
Pumping the decomposition fermentation mixed solution into a screw pump of 60m3In an anaerobic fermentation tank, the temperature of the fermentation tank is kept at 35 ℃, the liquid retention time is 30 days, and methane fermentation is carried out by a full mixed fermentation (CSTR) fermentation process to obtain a fermentation product.
The remaining dry matter in 1L of the fermentation product was measured by a dry matter loss method and the conversion rate ═ 100% (dry matter mass in 1L before fermentation-dry matter mass in 1L after fermentation)/dry matter mass in 1L before fermentation).
As a result, the conversion rate of the straw and the pig manure dry matter is more than 45 percent, and the daily biogas yield reaches 80m3The above.
Example 3 Combined methane fermentation of Rice straw and Chicken manure
1. Microbial inoculum for quickly decomposing organic matters
The organic matter fast decomposing microbial inoculum is prepared by the following method:
respectively culturing Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, Clostridium cellulolyticum (Xylanimomonas cellulolytica) DSM No.15894, Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 and Bacillus subtilis (Bacillus subtilis) ACCC No.03189 to obtain a Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555 culture solution, a Clostridium cellulolyticum (Xylanimidolytica) DSM No.15894 culture solution, a Pseudomonas sp (Pseudomonas sp.) ACCC No.11691 culture solution and a Bacillus subtilis (Bacillus subtilis) ACCC 03189 culture solution.
Mixing culture solution of Clostridium xylanolyticum (Clostridium xylanolyticum) DSM No.6555, culture solution of cellulolytic xylanase (Xanthomonas cellulolytica) DSM No.15894, culture solution of Pseudomonas sp (Pseudomonas sp.) ACCC No.11691, and culture solution of Bacillus subtilis ACCC No.03189, and obtaining mixed bacteria solution.
Crushed straw obtained by crushing dry rice straws and passing the crushed straw through a 0.4mm sieve is used as a carrier.
And then spraying a mixed bacterial liquid accounting for 10 percent of the mass of the chopped grass while stirring in 100Kg of chopped grass, and adsorbing the mixed bacterial liquid on a carrier to obtain the organic matter fast-decomposing microbial inoculum.
In the microbial inoculum, the mixture ratio of Clostridium xylanilyticum (Clostridium xylanilyticum), cellulolytic Pseudomonas (Xylanimonas cellulolytica), Pseudomonas (Pseudomonas sp.), Bacillus subtilis (Bacillus subtilis) and a carrier is 25cfu: 30cfu:30cfu:20 cfu: 5X 10-4g；
Wherein the content of Clostridium xylanisolvens (Clostridium xylanolyticum) in the microbial inoculum is 50000 cfu/g;
the content of cellulolytic xylan monad (Xylanimonas cellulosilytica) in the microbial inoculum is 60000 cfu/g;
the content of Pseudomonas (Pseudomonas sp.) in the microbial inoculum is 60000 cfu/g;
the content of the Bacillus subtilis in the microbial inoculum is 40000 cfu/g.
2. Combined methane fermentation of rice straw and chicken manure
The implementation place is as follows: \28095TheChina agriculture university scientific and technological park industry platform of Zhou city 60m3 full-mixing type
The components required for fermentation are as follows: rice straw (water content 12%), chicken manure (water content 78%), and the organic matter fast-decomposing microbial inoculum prepared in the step 1;
the implementation method comprises the following steps: effective volume of decomposition fermentation tank is 50m3The effective volume of the full-mixing anaerobic fermentation tank is 60m3。
The raw material ratio is as follows: 5.06 tons of rice straws, 2.5 tons of chicken manure and 42.5 tons of water are mixed evenly to obtain raw materials; the ratio of C to N of the raw materials is 20: 1;
then adding 5kg of the organic matter fast decomposing microbial inoculum prepared in the step 1 into 50.06 tons of raw materials;
crushing the straws into particles with the diameter less than 1cm, removing large particles with the diameter more than 1cm from excrement by a sieve, and adding the corn straws, the cow dung, the water and the microbial inoculum into an effective volume of 50m according to the proportion3The decomposition fermentation tank is stirred for 30 minutes at the rotating speed of 50 r/min and uniformly mixed, the temperature is controlled at 30 ℃, and the stirring intensity is controlled as follows: the rotation speed is 50 r/min, stirring is carried out for 10 min in each hour, stopping for 50 min to provide facultative anaerobic condition, and after running for 36 hours, 40% dry matter of the decomposing and fermenting system becomes water soluble, and 60% dry matter remains in suspension state to obtain decomposing and fermenting mixed liquor.
Pumping the decomposition fermentation mixed solution into a screw pump of 60m3In an anaerobic fermentation tank, the temperature of the fermentation tank is kept at 35 ℃, the liquid retention time is 30 days, and methane fermentation is carried out by a full mixed fermentation (CSTR) fermentation process to obtain a fermentation product.
Detecting the residual dry matter in the fermentation product 1L by a dry matter loss method, calculating the conversion rate (the dry matter mass in 1L before fermentation-the dry matter mass in 1L after fermentation)/the dry matter mass in 1L before fermentation) multiplied by 100 percent, and as a result, the conversion rate of the dry matters of the straws and the chicken manure is more than 50 percent, and the methane yield reaches 90m per day3The above.
The invention can also be matched according to the principle that the solid content in the raw material ratio is 10 percent and the C/N is equal to 35, and the operation conditions are the same as the 3 examples.

Claims (3)

1. A method of methane fermentation comprising the steps of: fermenting the raw materials by using a microbial inoculum to realize methane fermentation;
the raw materials consist of straws, animal wastes and water;
the mass ratio of the C element to the N element of the raw materials is 20: 1. 25:1 or 30: 1;
the mass ratio of the straw to the animal waste to the water to the microbial inoculum is 1: 1: 8: 0.001 or 1: 0.5: 8.5: 0.001;
the microbial inoculum is prepared by uniformly mixing a Clostridium xylanolyticum (Clostridium xylanolyticum) DSM number 6555 culture solution, a cellulolytic xylan monad (xylomonas cellulolytica) DSM No.15894 culture solution, a Pseudomonas (Pseudomonas sp.) ACCC No.11691 culture solution and a Bacillus subtilis ACCC No.03189 culture solution to obtain a mixed bacterial solution, and adsorbing the mixed bacterial solution onto a carrier to obtain the microbial inoculum;
in the microbial inoculum, the ratio of the Clostridium cellulolyticum (Clostridium cellulolyticum) DSM number 6555, the cellulolytic Pseudomonas (Xylanimonas cellulolytica) DSM No.15894, the Pseudomonas (Pseudomonas sp.) ACCC No.11691 and the Bacillus subtilis ACCC No.03189 to the carrier is (25-35) cfu: (25-30) cfu (20-30) cfu:20cfu: 5X 10-4g；
In the microbial inoculum, the content of Clostridium xylanolyticum (Clostridium xylolyticum) DSM number 6555 in the microbial inoculum is 50000-;
the content of the cellulolytic xylan monad (Xylanimonas cellulolysitica) DSM No.15894 in the microbial inoculum is 50000-60000 cfu/g;
the content of the Pseudomonas (Pseudomonas sp.) ACCC No.11691 in the microbial inoculum is 40000-;
the content of the Bacillus subtilis ACCC No.03189 in the microbial inoculum is 40000cfu/g;
the fermentation comprises the following steps:
1) uniformly mixing the microbial inoculum and the raw materials, stirring and fermenting to obtain a decomposition fermentation system;
2) fermenting the decomposition fermentation system in an anaerobic fermentation container to realize methane fermentation;
the stirring fermentation conditions are that the stirring speed is 50 r/min, stirring is carried out for 10 minutes in each hour, then the stirring is stopped for 50 minutes, the stirring fermentation is carried out for 36 hours, and the temperature is 30 ℃;
the anaerobic fermentation condition is 35 ℃ and 30 days.
2. The method of claim 1, wherein: the straws are corn straws, wheat straws or rice straws;
the straw is specifically straw particles smaller than 1 cm.
3. The method of claim 1, wherein: the animal waste is cow waste, pig waste or chicken waste.
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