CN102559499B - Preparation method of methane dry fermentation compound bacterial preparation - Google Patents

Preparation method of methane dry fermentation compound bacterial preparation Download PDF

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CN102559499B
CN102559499B CN201210021688.2A CN201210021688A CN102559499B CN 102559499 B CN102559499 B CN 102559499B CN 201210021688 A CN201210021688 A CN 201210021688A CN 102559499 B CN102559499 B CN 102559499B
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methane
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bacterium
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CN102559499A (en
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尹小波
李强
徐彦胜
马诗淳
张云飞
代丽蓉
承磊
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Biogas Institute of Ministry of Agriculture
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Abstract

The invention discloses a preparation method of a methane dry fermentation compound bacterial preparation, which comprises the following steps: mixing fermentation substrate with an inoculum which is compound bacteria composed of cellulose decomposing bacteria, proteolytic bacteria, fat decomposing bacteria, hydrogen-producing aceogenic bacteria, sulfate reducing bacteria and methanogenic archaea, adjusting the water content and pH of the mixed materials to 70-80% and 7.0 respectively, performing anaerobic fermentation under the condition of 35 DEG C plus or minus 2 DEG C for 60-90 days to obtain methane dry fermentation compound bacterial preparation. The preparation method of the invention can prepare compound bacterial preparation with a relatively complete methane fermentation function, and the total number of microbial cells and methanogenic archaea cells in the bacterial preparation are at least 10*10<10>/g and 1.0*10<8>/g respectively. Using the compound bacterial preparation as an inoculum of methane dry fermentation can reduce the starting time, and the fermentation process is stable and easy to control.

Description

A kind of preparation method of methane dry fermentation compound bacterial preparation
Technical field
The present invention relates to a kind of preparation method of methane dry fermentation compound bacterial preparation, belong to organic waste utilization and production of renewable energy resources technical field.
Background technology
In recent years, society constantly increases the demand of the energy, and the fossil energy resources such as oil, coal constantly reduce, and price continues soaring, bring disadvantageous effect to people's productive life, so the exploitation of renewable energy source are more and more subject to the attention of national governments and researcher.Biogas is clean renewable energy source, inexhaustible.The organism such as stalk, fowl and animal excrement, organic garbage of city can change into biogas through anaerobic digestion, for the mankind provide clean energy.
The microorganism that participates in biogas fermentation comprises zymogenic bacteria, hydrogen-producing acetogenic bacteria, the ancient bacterium of product methane etc.First, zymogenic bacteria is degraded to cellulose substances the organic acids such as acetic acid, propionic acid, by protein degradation, is organic acid and ammonia, by fat acid decomposition, is acetic acid, propionic acid, H 2and CO 2; Then, hydrogen-producing acetogenic bacteria resolves into acetic acid, H by the organic acid of generation 2and CO 2; Finally, methanogen is by H 2/ CO 2, the small-molecule substance such as methyl compound, acetic acid changes into methane, wherein the disappearance of any one link does not all reach methanogenic object.
Sludge gas dry fermentation is the focus of Recent study, it have fermentation solidity thing content high, save water, fermentation unit volume is little, fermentation after product is processed simple, the advantage that environmental pollution is little.But dry fermentation concentration is high, mass transfer is difficult, very high to the requirement of inoculum during startup.
According to pertinent literature, sludge gas dry fermentation starts the dewatered sludge, anaerobic activated sludge of fermented liquid that the inoculum adopt is mainly old methane-generating pit, cloaca mud, sewage work etc., the methane dry fermentation compound bacterial preparation of the special preparation of not mentioned use.
Chinese patent specification sheets CN101705199A discloses a kind of methane-producing composite microbial inoculum and preparation method, the method is utilized 5 strain methanogens to carry out liquid step by step as bacterial classification and after spreading cultivation, is mixed into a kind of methane-producing composite microbial inoculum, with mud or pig manure, adsorb again, to realize the production of solid composite fungus agent.This microbial inoculum can accelerate New-built Methane Tank or reload greatly after produce biogas start time, improve gas producing efficiency and the aerogenesis stability of methane-generating pit.This patent fungus strain forms single, the not mentioned startup for dry fermentation.
Chinese patent specification sheets CN101948752A discloses a kind of composite fungus agent for biogas fermentation, this microbial inoculum is a kind of compound formulation of microorganism zymocyte liquid, the microorganisms such as Mierocrystalline cellulose bacterioide, Pasteur's gemma clostridium fermented liquid, the bag-shaped bacterium of Black Sea methane, Ma Shi sarcina methanica have been comprised, mainly for take the conventional biogas fermentation that cow dung is main raw material, fermentation concentration is 5-10%.This microbial inoculum use range is narrow, also not mentioned for dry fermentation.
Chinese patent specification sheets CN101705199 discloses a kind of methane-producing composite microbial inoculum and preparation method thereof, this microbial inoculum by sarcina methanica, formic acid methagen, have a liking for tree methane tyrothricin, Ting Daer methane leaf bacterium, Ken Shi hair on the neck and send out methanobacteria and form, under anaerobic be inoculated in liquid methane bacterium culture medium, use this microbial inoculum can improve the gas production rate of normal operation methane-generating pit.This microbial inoculum only forms by producing the ancient bacterium of methane, there is no zymogenic bacteria and hydrogen-producing acetogens etc., and function singleness is not mentioned for dry fermentation.
Chinese patent specification sheets CN 101475926 and CN101481676 B disclose respectively a kind of anaerobic cellulose-degrading methane producing composite bacterium and making method.This composite bacteria comprises the 13 strain bacterium such as anaerobically fermenting bacterium, hydrogen-producing acetogens and methanogen.But its composition of the microorganism designs mainly for cellulosic degraded, the conventional biogas fermentation that is main raw material for rice straw, this microbial inoculum use range is narrow, also not mentioned for dry fermentation.
Summary of the invention
The object of the present invention is to provide a kind of preparation method who is applied to especially the composite fungus agent of sludge gas dry fermentation, according to the microorganism cells sum in the methane dry fermentation compound bacterial preparation of present method made, be at least 1.0 * 10 10individual/g, methanogen total cellular score is at least 1.0 * 10 8individual/g, can realize quick startup and steady running used as the inoculum of dry fermentation.
For achieving the above object, the technical solution used in the present invention is as follows:
A kind of preparation method of methane dry fermentation compound bacterial preparation, it is characterized in that: the composite bacteria that the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane is formed is as inoculum, inoculum is mixed with fermentation substrate, under 35 ± 2 ℃ of conditions, anaerobically fermenting, after 60 ~ 90 days, obtains methane dry fermentation compound bacterial preparation.
In described composite bacteria, the cellulose decomposing bacteria that is 25% ~ 45% by cell quantity per-cent, 9% ~ 15% proteolyticbacteria, 8% ~ 15% fat decomposing bacteria, 8% ~ 12% hydrogen-producing acetogenic bacteria, 18% ~ 31% sulphate reducing bacteria and 2.5% ~ 9.5% the ancient bacterium of product methane, the microorganism cells sum of each component is at least 2 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 5 * 10 8individual/g.
The described 11 strain bacterium that relate to as the composite bacteria of inoculum are as follows:
(1) bacteroides cellulosilyticusseparate Mierocrystalline cellulose bacterioide, strictly anaerobic, shaft-like, Gram-negative, the various cellulose substances of can degrading, can utilize various carbohydrates to generate acetic acid, propionic acid, succsinic acid, and bacterial strain deposit number is DSM 14838 t;
(2) clostridium sufflavumyellow bacillus fusiformis, strictly anaerobic, thalline is that microbend is shaft-like, can move, peritrichous, 33 ℃ of optimum growth temperatures, the most suitable growth pH 7.4, and energy degraded cellulose class material, can also utilize wood sugar, fructose, glucose, cellobiose, xylan, generates acetic acid, ethanol, H 2and CO 2, bacterial strain deposit number is DSM 19573 t;
(3) clostridium pasteurianumpasteur's gemma clostridium, 37 ℃ of optimum growth temperatures, do not move, and inferior utmost point end forms an oval gemma, can decomposing protein, carbohydrate, can glucose fermentation, maltose, lactose, sucrose produces acetic acid, bacterial strain deposit number is DSM 525 tor ATCC 6013 t;
(4) desulfobulbus rhabdoformispropionic acid desulfurization onion shape bacterium, strictly anaerobic, round bar shape, Gram-positive, 31 ℃ of optimum growth temperatures, optimal pH 6.8 ~ 7.2, degraded propionic salt is acetic acid and CO 2, also can utilize fumaric acid and oxysuccinic acid as carbon source and the energy, bacterial strain deposit number is DSM 8777 tor ATCC700652 t;
(5) syntrophomonas sapovoranslonger chain fatty acid syntrophism Zymomonas mobilis, strictly anaerobic, microbend is shaft-like, Gram-positive, 35 ℃ of optimum growth temperatures, the saturated chain lipid acid of lipid acid of 4 to 18 carbon atoms of energy oxygenolysis solution produces acetic acid and H 2, with methanospirillum hungateisyntrophism, bacterial strain deposit number is DSM 3441 t;
(6) syntrophomonas Wolfeiwo Shi syntrophism Zymomonas mobilis, strictly anaerobic, slight screw shaped, 35 ℃ of optimum growth temperatures, oxidation butyric acid produces acetic acid and H 2, and utilize H 2/ CO 2methanogen syntrophism, bacterial strain deposit number is DSM 4212 t;
(7) desulfovibrio desulfuricansdesulfovibrio desulfurican, strictly anaerobic, S bending, Gram-negative, polar flagella, can move, 37 ℃ of optimum growth temperatures, the vitriol of usining obtains energy as electron acceptor(EA) alienation organic substance, can metabolism lactic acid salt, the lipid acid such as pyruvic acid, and bacterial strain deposit number is DSM 642 t;
(8) methanospirillum hungateiheng Shi methanospirillum, strictly anaerobic, crooked shaft-like, 37 ℃ of optimum growth temperatures, optimal pH 7.0 ~ 7.5, can utilize H 2/ CO 2, formic acid produces CH 4, not utilizing acetic acid, methylamine, methyl alcohol and other alcohols, bacterial strain deposit number is DSM 864 t;
(9) methanocorpusculum parvumlittle methane grain bacterium, strictly anaerobic, Gram-negative, it is little irregular spherical that cell is, and 37 ℃ of optimum growth temperatures, can be with H 2/ CO 2, formate, propylene glycol/CO 2, butyleneglycol/CO 2for substrate produces methane, bacterial strain deposit number is DSM 3823 tor ATCC 43721 t;
(10) methanosarcina barkeripasteur's sarcina methanica, strictly anaerobic, gramstaining is variable, does not move, and is irregular spheroidal aggravation, single raw or typical many cells aggregate, 37 ℃ of optimum growth temperatures, can utilize methyl alcohol, Trimethylamine 99, acetic acid and H 2/ CO 2methane is produced in growth, but never utilizes formic acid, and bacterial strain deposit number is DSM 800 t;
(11) methanosaeta conciliimethane mane bacterium, strictly anaerobic, it is shaft-like that cell is, Gram-negative, 35 ℃ of optimum growth temperatures, growth pH6.5 ~ 7.8, only utilize acetate growth to produce methane, and bacterial strain deposit number is DSM 2139 t.
Above bacterial strain all can obtain by German microbial strains preservation center (DSMZ) or U.S. typical case culture center (ATCC).
Described solution Mierocrystalline cellulose bacterioide bacteroides cellulosilyticuswith yellow bacillus fusiformis clostridium sufflavum iscellulose decomposing bacteria; Described Pasteur's gemma clostridium clostridium pasteurianumfor proteolyticbacteria; Described longer chain fatty acid syntrophism Zymomonas mobilis syntrophomonas sapovoransfor fat decomposing bacteria; Described Wo Shi syntrophism Zymomonas mobilis syntrophomonas Wolfeifor hydrogen-producing acetogenic bacteria; Described desulfovibrio desulfurican desulfovibrio desulfuricanswith propionic acid desulfurization onion shape bacterium Desulfobulbus rhabdoformis be sulphate reducing bacteria; Described Heng Shi methanospirillum methanospirillum hungatei, little methane grain bacterium methanocorpusculum parvum, Pasteur's sarcina methanica methanosarcina barkeriwith methane mane bacterium methanosaeta conciliifor producing the ancient bacterium of methane.
The cell quantity degree of described solution Mierocrystalline cellulose bacterioide is 15% ~ 25%, the cell quantity degree of yellow bacillus fusiformis is 10% ~ 20%, the cell quantity degree of Pasteur's gemma clostridium is 9% ~ 15%, the cell quantity degree of propionic acid desulfurization onion shape bacterium is 10% ~ 15%, the cell quantity degree of longer chain fatty acid syntrophism Zymomonas mobilis is 8% ~ 15%, the cell quantity degree of Wo Shi syntrophism Zymomonas mobilis is 8% ~ 12%, the cell quantity degree of desulfovibrio desulfurican is 10% ~ 16%, the cell quantity degree of Heng Shi methanospirillum is 0.5% ~ 1%, the cell quantity degree of little methane grain bacterium is 1% ~ 2%, the cell quantity degree of Pasteur's sarcina methanica is 0.5% ~ 2.5%, the cell quantity degree of methane mane bacterium is 0.5% ~ 4%.
In described composite bacteria, microorganism cells sum is at least 2.0 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 5.0 * 10 8individual/g.
The mass ratio that described inoculum mixes with fermentation substrate is 3:10.
Described fermentation substrate can be selected pig manure, chicken manure, straw or maize straw, and ight soil raw material and stalk are in carbon nitrogen content ratio 20 ~ 30:1(mass ratio) be used in combination.
Described fermentation substrate should regulate TS content 20 ~ 30%, and under natural temperature condition, stack retting is 7 days.
Employing the invention has the advantages that:
One, microbial inoculum making method of the present invention is controlled at 20% ~ 30% fermentation concentration, has reduced the volume of fermentation unit, and cost is low, and Production Flow Chart is easy to control.
Two, in the present invention, with the dry fermenting compound fungus of high density, make inoculum, using dosage is little, with low cost, simple to operate.
three,in preparation method of the present invention, composite fungus agent is comprised of 11 strain anaerobism function stems, its physiological property is clear, by function, classifies, and belongs to respectively the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane; In methane dry fermentation compound bacteria formula, the bacterial classification mixed culture of many strains definite functions, produces enzyme abundant, can remove the feedback inhibition of meta-bolites, ensures the stability of yeasting.While using it for sludge gas dry fermentation, can significantly shorten start time, improve biogas fermentation efficiency, strengthen the stability of fermenting process.
Four, in the present invention, the ratio of the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane in microbial inoculum as inoculum can, according to adjusting in the described scope that do not coexist of fermentation materials, make it to reach better result of use.
Five, in the present invention, by inoculum, than controlling, be 3:10 with the mixing quality of fermentation substrate, sufficient nutrient can be provided at the growth metabolism that the fermentation starting stage is bacterial classification, guarantee fast, stably to enter the bacteria fermentation stage, realize the rapid propagation of microorganism.
Six, in the present invention as pig manure, cow dung, chicken manure, straw or the maize straw of fermentation substrate, there is carbon-nitrogen ratio clear and definite, easily collecting, with low cost, the advantage that ferment effect is stable.
Seven, in the present invention, reasonably combined by different types of raw material, can guarantee that in fermentation substrate, carbon nitrogen content scale dimension is held in 20 ~ 30:1, for microorganism provides sufficient Carbon and nitrogen sources in growth metabolism process, can, for microorganism provides stable growing environment, guarantee that microorganism breeds fast.
Eight, utilize method related in the present invention can obtain a kind of methane dry fermentation compound bacterial preparation of high-biomass, the microorganism cells sum of microbial inoculum is at least 1.0 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 1.0 * 10 8individual/g; This microbial inoculum moisture content is low, is easy to store, convenient transportation, using method be easy to operate.
Embodiment
Embodiment 1
Composite bacteria of the present invention is comprised of 11 strain bacterium, according to the difference of function in fermenting process, this 11 strain bacterium can be divided into following 6 classes:
Cellulose decomposing bacteria: bacteroides cellulosilyticus, clostridium sufflavum;
Proteolyticbacteria: clostridium pasteurianum;
Fat decomposing bacteria: syntrophomonas sapovorans;
Sulphate reducing bacteria: desulfovibrio desulfuricans, desulfobulbus rhabdoformis;
Hydrogen-producing acetogenic bacteria: syntrophomonas Wolfei;
Produce the ancient bacterium of methane: methanospirillum hungatei, methanocorpusculum parvum, methanosarcina barkeri, methanosaeta concilii.
Above-mentioned 11 strain bacterium of adhering to the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane separately are directly mixed and form composite bacteria as inoculum in proportion, and the organism such as pig manure, chicken manure, straw or maize straw of take are fermentation substrate.Ight soil raw material and stalk can be used in combination in carbon nitrogen content ratio 20 ~ 30:1, for example ight soil raw material and stalk are pressed 20:1,25:1 or 30:1.By inoculum with fermentation substrate by 3:10(mass ratio) mix after, regulating TS content is 20% ~ 30%, for example, regulate TS content to 20%, 25% or 30%, pH value 6.5 ~ 7.0, at 35 ± 2 ℃, anaerobism cultivation is 60 ~ 90 days.In said process, 11 strain function yeast synergies in inoculum, the substrate of degrading generates methane, and each function stem is realized propagation simultaneously, obtains the methane dry fermentation compound bacterial preparation of a kind of composition and stable performance.In this microbial inoculum, microorganism cells sum content can reach 1.0 * 10 10individual/g, produces the ancient mycetocyte sum of methane and can reach 1.0 * 10 8individual/g.
The content of dry-matter in the methane dry fermentation compound bacterial preparation that in embodiments of the invention, " TS content (%) " refers to produce; " total cellular score (individual/gram) " refers in every gram of above-mentioned microbial inoculum the cell quantity containing 11 kinds of functional microorganisms; " methanogen cell count (individual/gram) " refer in every gram of above-mentioned microbial inoculum containing the cell quantity that produces the ancient bacterium of methane; PH value refers to the pH value of the methane dry fermentation compound bacterial preparation of acquisition.
The cell quantity degree of described solution Mierocrystalline cellulose bacterioide is 15% ~ 25%, the cell quantity degree of yellow bacillus fusiformis is 10% ~ 20%, the cell quantity degree of Pasteur's gemma clostridium is 9% ~ 15%, the cell quantity degree of propionic acid desulfurization onion shape bacterium is 10% ~ 15%, the cell quantity degree of longer chain fatty acid syntrophism Zymomonas mobilis is 8% ~ 15%, the cell quantity degree of Wo Shi syntrophism Zymomonas mobilis is 8% ~ 12%, the cell quantity degree of desulfovibrio desulfurican is 10% ~ 16%, the cell quantity degree of Heng Shi methanospirillum is 0.5% ~ 1%, the cell quantity degree of little methane grain bacterium is 1% ~ 2%, the cell quantity degree of Pasteur's sarcina methanica is 0.5% ~ 2.5%, the cell quantity degree of methane mane bacterium is 0.5% ~ 4%.
Embodiment 2
In inoculum, the proportion of composing of 11 strain bacterium is specific as follows: bacteroides cellulosilyticusaccount for 22%, clostridium sufflavumaccount for 17%, clostridium pasteurianumaccount for 10%, desulfobulbus rhabdoformisaccount for 11%, syntrophomonas sapovoransaccount for 11%, syntrophomonas Wolfeiaccount for 10%, desulfovibrio desulfuricansaccount for 13%, methanospirillum hungateiaccount for 0.5%, methanocorpusculum parvumaccount for 1.0%, methanosarcina barkeriaccount for 1.5%, methanosaeta conciliiaccount for 3%.
Mixture with pig manure and maize straw is made fermentation substrate.Corn stalk powder is broken into powder, with pig manure in mass ratio the ratio of 1:1 mix, adding water, to regulate TS content be 30%, under natural temperature, stack retting is 7 days.After stack retting finishes, with lime powder, regulate pH value to 7.0, then by raw material: inoculum=10:3(mass ratio) add above-mentioned inoculum, after mixing, the moisture content to 80% of adjustable compound, anaerobism cultivation under 35 ± 2 ℃ of conditions.After 60 ~ 90 days, obtain composite fungus agent, its indices is as shown in the table:
Embodiment 3
Inoculum is comprised of following 11 strain bacterium, bacteroides cellulosilyticusaccount for 20%, clostridium sufflavumaccount for 15%, clostridium pasteurianumaccount for 12%, desulfobulbus rhabdoformisaccount for 11%, syntrophomonas sapovoransaccount for 13%, syntrophomonas Wolfeiaccount for 10%, desulfovibrio desulfuricansaccount for 13%, methanospirillum hungateiaccount for 1.0%, methanocorpusculum parvumaccount for 1.5%, methanosarcina barkeriaccount for 2%, methanosaeta conciliiaccount for 1.5%.
Mixture with pig manure and rice straw is made fermentation substrate.Rice straw is ground into powder, with pig manure in mass ratio the ratio of 1:1 mix, adding water, to regulate TS content be 25%, under natural temperature, stack retting is 7 days.After stack retting finishes, with lime powder, regulate pH value to 6.5, then by fermentation substrate: inoculum=10:3(mass ratio) add above-mentioned inoculum.After mixing, under 35 ± 2 ℃ of conditions, anaerobism is cultivated.After 60 ~ 90 days, obtain composite fungus agent, its indices is as shown in the table:
Embodiment 4
Inoculum is comprised of following 11 strain bacterium, bacteroides cellulosilyticusaccount for 19%, clostridium sufflavumaccount for 14%, clostridium pasteurianumaccount for 14%, desulfobulbus rhabdoformisaccount for 11%, syntrophomonas sapovoransaccount for 13%, syntrophomonas Wolfeiaccount for 10%, desulfovibrio desulfuricansaccount for 13%, methanospirillum hungateiaccount for 0.5%, methanocorpusculum parvumaccount for 1.5%, methanosarcina barkeriaccount for 2%, methanosaeta conciliiaccount for 2%.
Mixture with chicken manure and rice straw is made fermentation substrate.Rice straw is ground into powder, with chicken manure in mass ratio the ratio of 1:1 mix, adding water, to regulate TS content be 20%, under natural temperature, stack retting is 7 days.After stack retting finishes, with lime powder, regulate pH value to 7.0, then by fermentation substrate: inoculum=10:3(mass ratio) add above-mentioned inoculum.After mixing, under 35 ± 2 ℃ of conditions, anaerobism is cultivated.After 60 ~ 90 days, obtain composite fungus agent, its indices is as shown in the table:
Embodiment 5
Inoculum is comprised of following 11 strain bacterium, bacteroides cellulosilyticusaccount for 18%, clostridium sufflavumaccount for 14%, clostridium pasteurianumaccount for 15%, desulfobulbus rhabdoformisaccount for 11%, syntrophomonas sapovoransaccount for 13%, syntrophomonas Wolfeiaccount for 10%, desulfovibrio desulfuricansaccount for 13%, methanospirillum hungateiaccount for 1.0%, methanocorpusculum parvumaccount for 1.5%, methanosarcina barkeriaccount for 1.5%, methanosaeta conciliiaccount for 2%.
Mixture with chicken manure and maize straw is made fermentation substrate.Corn stalk powder is broken into powder, with chicken manure in mass ratio the ratio of 1:1 mix, adding water, to regulate TS content be 20%, under natural temperature, stack retting is 7 days.After stack retting finishes, with lime powder, regulate pH value to 7.0, then by fermentation substrate: inoculum=10:3(mass ratio) add above-mentioned inoculum.After mixing, under 35 ± 2 ℃ of conditions, anaerobism is cultivated.After 60 ~ 90 days, obtain composite fungus agent, its indices is as shown in the table:
Embodiment 6
A kind of preparation method of methane dry fermentation compound bacterial preparation, the composite bacteria that the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane is formed is as inoculum, inoculum is mixed with fermentation substrate, under 33 ℃ of conditions, carry out anaerobically fermenting, ferment after 90 days, obtain methane dry fermentation compound bacterial preparation.
In described composite bacteria of the present invention, the cellulose decomposing bacteria that is 25% by cell quantity per-cent, 15% proteolyticbacteria, 15% fat decomposing bacteria, 8% hydrogen-producing acetogenic bacteria, 27.5% sulphate reducing bacteria and 9.5% the ancient bacterium of product methane, the microorganism cells sum of each component is at least 2 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 5 * 10 8individual/g.
Described cellulose decomposing bacteria can be for separating Mierocrystalline cellulose bacterioide bacteroides cellulosilyticusand/or yellow bacillus fusiformis clostridium sufflavum; Described proteolyticbacteria can be Pasteur's gemma clostridium clostridium pasteurianum; Described fat decomposing bacteria can be longer chain fatty acid syntrophism Zymomonas mobilis syntrophomonas sapovorans; Described hydrogen-producing acetogenic bacteria can be Wo Shi syntrophism Zymomonas mobilis syntrophomonas wolfei; Described sulphate reducing bacteria is desulfovibrio desulfurican desulfovibrio desulfuricansand/or propionic acid desulfurization onion shape bacterium desulfobulbus rhabdoformis; The ancient bacterium of described product methane can be Heng Shi methanospirillum methanospirillum hungatei, little methane grain bacterium methanocorpusculum parvum, Pasteur's sarcina methanica methanosarcina barkeriand/or methane mane bacterium methanosaeta concilii.Those skilled in the art can select in above-mentioned bacterium, but is not limited to above-mentioned bacterium.
Embodiment 7
A kind of preparation method of methane dry fermentation compound bacterial preparation, the composite bacteria that the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane is formed is as inoculum, inoculum is mixed with fermentation substrate, under 37 ℃ of conditions, carry out anaerobically fermenting, ferment after 60 days, obtain methane dry fermentation compound bacterial preparation.
In described composite bacteria, the cellulose decomposing bacteria that is 45% by cell quantity per-cent, 9% proteolyticbacteria, 8% fat decomposing bacteria, 12% hydrogen-producing acetogenic bacteria, 18% sulphate reducing bacteria and 8% the ancient bacterium of product methane, the microorganism cells sum of each component is at least 2 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 5 * 10 8individual/g.
Embodiment 8
A kind of preparation method of methane dry fermentation compound bacterial preparation, the composite bacteria that the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane is formed is as inoculum, inoculum is mixed with fermentation substrate, under 35 ℃ of conditions, carry out anaerobically fermenting, ferment after 80 days, obtain methane dry fermentation compound bacterial preparation.
In described composite bacteria, the cellulose decomposing bacteria that is 30% by cell quantity per-cent, 12% proteolyticbacteria, 12% fat decomposing bacteria, 10% hydrogen-producing acetogenic bacteria, 31% sulphate reducing bacteria and 5% the ancient bacterium of product methane, the microorganism cells sum of each component is at least 2 * 10 10individual/g, produces the ancient mycetocyte sum of methane and is at least 5 * 10 8individual/g.

Claims (2)

1. a preparation method for methane dry fermentation compound bacterial preparation, is characterized in that: first regulate fermentation substrate TS content 20 ~ 30%, under natural temperature condition, stack retting is 7 days; The composite bacteria that the ancient bacterium of cellulose decomposing bacteria, proteolyticbacteria, fat decomposing bacteria, hydrogen-producing acetogenic bacteria, sulphate reducing bacteria and product methane is formed is as inoculum, after stack retting finishes, by inoculum and fermentation substrate in mass ratio 3:10 mix, under 35 ± 2 ℃ of conditions, carry out anaerobically fermenting, ferment after 60 ~ 90 days, obtain methane dry fermentation compound bacterial preparation;
Described cellulose decomposing bacteria is for separating Mierocrystalline cellulose bacterioide bacteroides cellulosilyticuswith yellow bacillus fusiformis clostridium sufflavum; Described proteolyticbacteria is Pasteur's gemma clostridium clostridium pasteurianum; Described fat decomposing bacteria is longer chain fatty acid syntrophism Zymomonas mobilis syntrophomonas sapovorans; Described hydrogen-producing acetogenic bacteria is Wo Shi syntrophism Zymomonas mobilis syntrophomonas wolfei; Described sulphate reducing bacteria is desulfovibrio desulfurican desulfovibrio desulfuricanswith propionic acid desulfurization onion shape bacterium desulfobulbus rhabdoformis; The ancient bacterium of described product methane is Heng Shi methanospirillum methanospirillum hungatei, little methane grain bacterium methanocorpusculum parvum, Pasteur's sarcina methanica methanosarcina barkeriwith methane mane bacterium methanosaeta concilii;
The quantity degree of described solution Mierocrystalline cellulose bacterioide is 15 ~ 25%, the quantity degree of yellow bacillus fusiformis is 10 ~ 20%, the quantity degree of Pasteur's gemma clostridium is 9 ~ 15%, the quantity degree of propionic acid desulfurization onion shape bacterium is 10 ~ 15%, the quantity degree of longer chain fatty acid syntrophism Zymomonas mobilis is 8 ~ 15%, the quantity degree of Wo Shi syntrophism Zymomonas mobilis is 8 ~ 12%, the quantity degree of desulfovibrio desulfurican is 10 ~ 16%, the quantity degree of Heng Shi methanospirillum is 0.5 ~ 1%, the quantity degree of little methane grain bacterium is 1 ~ 2%, the quantity degree of Pasteur's sarcina methanica is 0.5 ~ 2.5%, the quantity degree of methane mane bacterium is 0.5 ~ 4%.
2. the preparation method of a kind of methane dry fermentation compound bacterial preparation according to claim 1, is characterized in that: described fermentation substrate is pig manure, chicken manure, straw or maize straw, and ight soil raw material and stalk are used in combination in carbon nitrogen content ratio 20 ~ 30:1.
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