WO2010072220A1 - Paenibacillus macerans pour traiter de la biomasse - Google Patents

Paenibacillus macerans pour traiter de la biomasse Download PDF

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Publication number
WO2010072220A1
WO2010072220A1 PCT/DE2009/075036 DE2009075036W WO2010072220A1 WO 2010072220 A1 WO2010072220 A1 WO 2010072220A1 DE 2009075036 W DE2009075036 W DE 2009075036W WO 2010072220 A1 WO2010072220 A1 WO 2010072220A1
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Prior art keywords
microorganisms
microorganism
biomass
culture
paenibacillus macerans
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PCT/DE2009/075036
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German (de)
English (en)
Inventor
Monika Reuter
Daniel Vater
Vera Duchow
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Schmack Biogas Ag
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Priority claimed from PCT/DE2008/075016 external-priority patent/WO2009086811A2/fr
Priority claimed from DE102009003587A external-priority patent/DE102009003587A1/de
Application filed by Schmack Biogas Ag filed Critical Schmack Biogas Ag
Priority to PCT/DE2010/075019 priority Critical patent/WO2010102618A2/fr
Publication of WO2010072220A1 publication Critical patent/WO2010072220A1/fr

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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
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
    • C12P5/023—Methane
    • 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
    • C10—PETROLEUM, GAS OR COKE INDUSTRIES; TECHNICAL GASES CONTAINING CARBON MONOXIDE; FUELS; LUBRICANTS; PEAT
    • C10L—FUELS NOT OTHERWISE PROVIDED FOR; NATURAL GAS; SYNTHETIC NATURAL GAS OBTAINED BY PROCESSES NOT COVERED BY SUBCLASSES C10G, C10K; LIQUEFIED PETROLEUM GAS; ADDING MATERIALS TO FUELS OR FIRES TO REDUCE SMOKE OR UNDESIRABLE DEPOSITS OR TO FACILITATE SOOT REMOVAL; FIRELIGHTERS
    • C10L3/00—Gaseous fuels; Natural gas; Synthetic natural gas obtained by processes not covered by subclass C10G, C10K; Liquefied petroleum gas
    • C10L3/06—Natural gas; Synthetic natural gas obtained by processes not covered by C10G, C10K3/02 or C10K3/04
    • C10L3/08—Production of synthetic natural gas
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/145—Clostridium
    • 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
    • Y02E50/10—Biofuels, e.g. bio-diesel
    • 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

Definitions

  • the invention relates to a method for the treatment of biomass using a microorganism of the species Paenibacillus macerans.
  • Biofuels are mainly obtained by fermentation of plant substrates with the help of yeasts, bacteria or fungi.
  • the produced liquid energy carriers in particular bioethanol, can then be used as fuel in suitable combustion plants or as fuel or additive in motor vehicle engines or motors for power and heat generation.
  • the biomass is thus converted into a liquid and easily transportable energy source with a relatively high energy density, which can then be used universally.
  • the plant material used as raw material and in particular the organic dry substance contained therein must be opened for a subsequent utilization. This is done by hydrolysis of the organic dry matter, which also corresponds to a liquefaction of the organic dry matter. Ethanol or another biofuel is then produced from the liquid biomass substrate by fermentation.
  • oTS organic dry substance contained therein
  • the ethanolic fermentation takes place primarily by added yeasts, which convert glucose into ethanol.
  • yeasts which convert glucose into ethanol.
  • firstly high molecular weight substrates such as starch, cellulose or hemicellulose (for example in cereals, straw, whole plants) must be enzymatically or chemically cleaved to allow alcoholic fermentation.
  • hydrolytic substrate digestion which also corresponds to liquefaction, are primarily responsible microorganisms, especially bacteria.
  • biogas plants methane is produced by a microbial decomposition process of organic substances.
  • the biogas is produced in a multi-stage process of fermentation or digestion by the activity of anaerobic or microaerophilic microorganisms, i. in the absence of air.
  • the organic material used as a fermentation substrate has, from a chemical point of view, a high molecular structure, which in the individual process steps of a Biogas plant is degraded by metabolic activity of microorganisms to low molecular weight building blocks.
  • the populations of microorganisms which are active in the fermentation of the organic fermentation substrate have hitherto been insufficiently characterized.
  • exoenzymes e.g., cellulases, amylases, proteases, lipases
  • exoenzymes e.g., cellulases, amylases, proteases, lipases
  • the gaseous products formed besides consist predominantly of carbon dioxide.
  • hydrolysis products eg mono-, disaccharides, di-, oligopeptides, amino acids, glycerol, long chain fatty acids
  • short chain fatty or carboxylic acids such as butter -, propionic and acetic acid
  • short-chain alcohols such as ethanol
  • the short-chain fatty acids and carboxylic acids formed in acidogenesis and the short-chain alcohols are taken up by acetogenic bacteria and excreted again as acetic acid after ⁇ -oxidation.
  • By-products of acetogenesis are CO 2 and molecular hydrogen (H 2 ).
  • the products of acetogenesis such as acetic acid but also other substrates such as methanol and formate are converted by methane-forming organisms in the obligate anaerobic methanogenesis to methane and CO 2 .
  • the resulting here CO 2 and also during the other process steps such as hydrolysis, CO 2 formed in turn can also be converted by microorganisms with the incurred H 2 to methane.
  • the volume loading of a fermenter is the amount of substrate fed to the fermenter, expressed in kilograms of dry organic matter per cubic meter of fermenter volume and per day.
  • the amount of biogas produced depends strongly on the volume load of the fermenter, with increasing space load an increasingly larger amount of biogas is generated.
  • a high space load makes the process of biogas production increasingly economically viable, but on the other hand leads to an increasing destabilization of the biological processes of fermentation.
  • Increasing the volume of space is one way to operate a biogas plant more efficiently.
  • the biogas production is increased by a slow increase in the amount of feed in oTS per day.
  • high space loads of more than 6 kgoTS / m 3 d are not yet available.
  • the goal in plant operation is to operate a plant economically.
  • the stability of the biological process is at the forefront, as plant failure causes extremely high costs.
  • Substrates for biogas production have dry matter contents in the range of 5% to 90%.
  • substrates with a dry matter content of up to about 35 to at most 40% can be used. Since substrates with a higher dry matter content, in particular a higher proportion of organic dry matter, usually provide a higher energy content, they would preferably be used. In this case, however, there are higher costs due to increased expenditure of energy during pumping and stirring as well as higher ancillary costs due to wear or repair of pumps, agitators or the like with high viscosity or a high dry matter content. If substrate dilution is necessary because of a high dry matter content, additional water and wastewater costs as well as technical equipment for water supply, recovery or wastewater treatment are incurred.
  • biofuel is understood as meaning a liquid or gaseous fuel or energy carrier which is produced from biomass.
  • Biofuels come for the operation of Internal combustion engines for both mobile (eg motor vehicles) and stationary (eg generation of electrical and thermal energy in a combined heat and power plant) applications are used.
  • Examples of biofuels are biodiesel, bioethanol, biomethanol, biokerosene, biohydrogen or biogas.
  • bioethanol refers to ethanol which has been produced by alcoholic fermentation from biomass as renewable carbon carrier or biodegradable fractions of waste and serves in various concentrations as an additive to mineral oil fuels such as biodiesel or biofuel.
  • biogas is understood to mean the gaseous product of the anaerobic biodegradation of organic substrates, which generally contains about 45-70% of methane, 30-55% of carbon dioxide, and small amounts of nitrogen, hydrogen sulphide and other gases.
  • fermentation or “fermentation” in the context of the present invention include both anaerobic and aerobic metabolic processes which, under the action of microorganisms in a technical process from the supplied substrate to produce a product, e.g. Lead biogas.
  • a differentiation from the term “fermentation” is given in that it is exclusively anaerobic processes.
  • a “fermenter” is understood to mean the container in which the microbiological degradation of the substrate takes place with simultaneous formation of biogas.
  • the terms “reactor”, “fermenter” and “digester” are used interchangeably.
  • fertilization substrate or "substrate” in the context of the present invention means organic and biodegradable material which is added to the fermenter for fermentation.
  • Substrates can be renewable raw materials, organic fertilizers, substrates from the processing agricultural industry, municipal organic residues, slaughter residues or green waste.
  • substrates are corn silage, rye silage, Beet pulp, molasses, grass silage, cattle or pig slurry, beef, pork, chicken or horse manure, beer grains, apple, fruit or vine pomace, cereal, potato or fruit vinasse.
  • the terms “fermentation substrate” and “substrate” are used synonymously.
  • fertilization residue or "digestate” is understood to be the residue of biogas production which leaves the fermenter and is frequently stored in its own container.
  • volume load is understood to mean the amount of dry organic matter (oTS) in kg supplied to the fermenter per day and cubic meter (m 3 ) working volume.
  • organic dry substance (oTS) is understood to mean the anhydrous organic fraction of a substance mixture after removal of the inorganic constituents and drying at 105 ° C. As a rule, the dry matter content is stated in% of the substrate.
  • the term “residence time” is understood to mean the average residence time of the substrate in the fermenter.
  • specific biogas yield or “specific methane yield” is used to denote the amount of biogas or methane produced (stated in standard cubic meters of Nm 3 gas) divided by the amount (as a rule per tonne) of organic dry substance used or substrate understood.
  • nucleotide sequence encompasses both the DNA sequence and the corresponding RNA sequence,
  • RNA sequences given in the invention using bases A, U, C, G also refer to the corresponding DNA sequences using bases A, T, C, G and vice versa.
  • nucleotide sequence of a microorganism by means of a standardized process by which the individual nucleotides of the DNA or RNA of the microorganism can be detected with high accuracy.
  • individual positions can repeatedly be present in a sequence for which the determination of the nucleotide present at the respective position was not possible with sufficient accuracy.
  • the letter "N" is indicated in the nucleotide sequence in the context of the present invention. If the letter "N" is subsequently used in a nucleotide sequence, this stands as an abbreviation for every conceivable nucleotide, that is to say for A, U, G or C. in the case of a ribonucleic acid or for A, T, G or C in the case of a deoxyribonucleic acid.
  • nucleotide mutation means a change in the starting nucleotide sequence, whereby individual nucleotides or several, directly following one another or interrupted by unmodified nucleotides
  • insertion or “addition” as used herein means the addition of 1, 2 or more nucleotides to the respective starting sequence.
  • substitution means the replacement of a nucleotide present at a particular position by another.
  • microorganism is understood to mean microscopically small organisms, which as a rule are single-celled organisms but may also be multicellular organisms Examples of microorganisms are bacteria, microscopic algae, fungi or protozoa.
  • the term "genus” of microorganisms, "type” of microorganisms and “strain” of microorganisms is understood to mean the corresponding basic category of biological taxonomy, in particular the phylogenetic classification Among a particular genus, species or strain, not only are microorganisms having a particular RNA sequence but also, to a certain extent, their genetic variants, with genetic variance in the strain, species, genus increases.
  • culture refers to an accumulation of microorganisms under established conditions which ensure the growth or at least the survival of the microorganisms, for example enrichment cultures or pure cultures, liquid cultures as well as cultures on solid media such as nutrient media also permanent crops such as frozen glycerin cultures, immobilized cultures such as gel cultures or highly concentrated cultures such as cell pellets.
  • pure culture of a microorganism is understood to mean the progeny of a single cell, which is isolated by a multi-step process from a mixture of different microorganisms.
  • the multi-step process involves the separation of a single cell from a cell population and requires that also from the cell
  • pure cultures of microorganisms can be selectively recovered
  • serial dilution of the suspension in the nutrient solution it can finally be achieved that in the last dilution stage there is only one cell left then the basis for a pure culture.
  • mixed culture is understood to mean a mixture of different microorganisms, but natural populations of microorganisms are usually mixed cultures, but mixed cultures can also be produced artificially, for example by combining several pure cultures.
  • the object of the invention is to provide a method for the treatment of biomass, which allows a higher yield of usable biofuels than the prior art.
  • the present invention provides a method of treating biomass.
  • the biomass is added to a microorganism of the species Paenibacillus macerans.
  • the method for treating biomass is preferably a method for liquefying biomass.
  • Paenibacillus macerans is known as an optionally anaerobically living organism.
  • the use or occurrence of microorganisms of the species Paenibacillus macerans in the treatment of biomass or the production of biogas was previously unknown.
  • the addition of microorganisms of the species Paenibacillus macerans to a biomass substrate the viscosity of the substrate can be greatly reduced.
  • the use of microorganisms of the species Paenibacillus macerans leads to a significant increase in the liquefaction of the biomass and, as a result, to a significant improvement in the efficiency and efficiency of biofuel production plants.
  • the method for the treatment of biomass is a method for producing biogas from biomass.
  • the addition of microorganisms of the species Paenibacillus macerans to the fermentation substrate can both increase the volume load of the fermenter and significantly increase the amount of biogas formed.
  • the addition of a microorganism of the species Paenibacillus macerans causes an increase in the volume load of a fermenter up to more than 50%, without any instability of the fermentation process would occur. Parallel to the increased space load, the amount of biogas produced is significantly increased.
  • the specific yield of biogas increases, since significantly more of the organic dry matter is degraded than in the absence of addition of microorganisms of the species Paenibacillus macerans. Due to the increased degree of degradation, a significantly increased specific gas yield can be achieved with improved substrate utilization. In addition, by the Addition of microorganisms of the species Paenibacillus macerans the residence time of the fermentation substrate in the fermenter at constant gas yield can be significantly shortened, whereby the increase in the space load is possible. The use of microorganisms of the species Paenibacillus macerans therefore leads to a dramatic improvement in the efficiency and efficiency of biogas plants.
  • the inventive addition of microorganisms of the species Paenibacillus macerans thus provides a method which ensures increased stability of the fermentation process and in which liquefaction of the fermentation substrate occurs.
  • the liquefaction of substrate by the decomposition of insoluble constituents can also be used for the production of biofuel from biomass.
  • a microorganism of the species Paenibacillus macerans is added in the form of a culture of microorganisms consisting predominantly of a microorganism of the species Paenibacillus macerans persists.
  • microorganisms of the species Paenibacillus macerans could only be detected in minute traces of less than 10% 4 % of the total number of microorganisms present, since the amount of microorganisms isolated from their natural occurrence is generally sufficient for the addition of the microorganisms
  • the addition of the microorganisms to the fermentation substrate of a fermenter is most easily carried out directly in the form of a culture of microorganisms.
  • the addition of the culture of Paenibacillus macerans can be carried out in the form of a culture suspension, in the form of dry, freeze-dried or moist cell pellets or also in the form of spore suspensions, spore preparations or dry, freeze-dried or moist spore pellets.
  • Microorganisms of the species Clostridium sartagoformum and Clostridium sporosphaeroides have similar properties with respect to the treatment of biomass as the microorganisms of the species Paenibacillus macerans, which is why they are predestined for use in the treatment of biomass. Microorganisms of the species Clostridium sartagoformum and Clostridium Sporosphaeroides can therefore also be used in the methods and applications described herein for the treatment of biomass.
  • Microorganisms of the species Paenibacillus macerans are preferably added to the fermentation substrate in the form of cultures of microorganisms, the cultures of microorganisms consisting predominantly of microorganisms of the species Paenibacillus macerans. If, in addition to the determination of the number of microorganisms of the species Paenibacillus macerans, the total number of microorganisms is also determined, the proportion of microorganisms of the species Paenibacillus macerans in the culture may be expressed as a percentage. In a mixed culture, microorganisms of the species Paenibacillus macerans are the predominant species of microorganisms when they have the highest percentage of the various types of microorganisms present in the mixed culture.
  • composition of the microbial populations in the various fermentation substrates as well as the development of the organism composition during the fermentation process is largely unknown, but very variable and subject to a complicated dynamic process, which is also influenced by the respective process conditions.
  • various methods are known to those skilled in the art, for example, in the review article by Amann et al. (Microbiol. Review., 59, 143-169, 1995).
  • a preferred method for determining the microorganism composition independently of a previous cultivation of the microorganisms is, for example, the preparation of a rDNA clone library (eg based on 16S rRNA) after nucleic acid extraction and PCR, which can then be sequenced.
  • a rDNA clone library eg based on 16S rRNA
  • the composition of the microbial population in the fermentation substrate can be determined, for example, by in situ hybridization with specific fluorescence-labeled oligonucleotide probes.
  • Suitable rRNA-based oligonucleotide probes are known from the review mentioned above or may be prepared, for example, by probe base (Loy et al., 2003, Nucleic Acids Res. 31, 514-516.
  • a quantitative determination of the proportion of individual microorganisms in the total population can be carried out in a suitable manner with the methods of quantitative dot blot, in situ hybridization or whole cell hybridization.
  • the microorganism Paenibacillus macerans accounts for at least 10 "4 % of the total number of microorganisms present in the culture added to the fermentation substrate.” More preferably, the microorganism Paenibacillus macerans accounts for at least 10 "2 % of the total number of microorganisms present in the culture and particularly preferably, the microorganism Paenibacillus macerans accounts for at least 1% of the total number of microorganisms present in the culture.
  • the microorganism Paenibacillus macerans accounts for at least 10% of the total number of microorganisms present in the culture, more preferably the microorganism Paenibacillus macerans accounts for at least 50% of the total number of microorganisms present in the culture, and more preferably the microorganism makes Paenibacillus macerans at least 90% of the total number of microorganisms present in the culture.
  • a pure culture of a microorganism of the species Paenibacillus macerans is added.
  • the pure culture is biochemically characterized by specific metabolic processes and activities, as well as by special growth conditions. Due to the specific metabolic processes and activities, the addition of a pure culture of a fermentative microorganism can especially contribute to an improved control of the complex biogas production process.
  • a microorganism of the species Paenibacillus macerans as a component at least added to an immobilized culture of microorganisms. Since the amount of microorganisms isolated from their natural occurrence is insufficient for the addition of the microorganisms, it is usually propagated in the form of a culture. In practice, it has been found that the addition of the microorganisms to the fermentation substrate of a fermenter is most easily carried out in the form of an immobilized culture of microorganisms.
  • microorganisms of the species Paenibacillus macerans should be present in the added immobilized culture in an amount enriched in comparison to the natural occurrence.
  • immobilized mixed cultures of any composition can be used for the addition. The only requirement is that microorganisms of the species Paenibacillus macerans are present in an amount that exceeds their natural occurrence.
  • the microorganism of the species Paenibacillus macerans accounts for at least 10.sup.- 4 % of the total number of microorganisms present in the immobilized culture added to the fermentation substrate.
  • the microorganism of the species Paenibacillus macerans accounts for at least 10.sup.- 2 % of the total number In microorganisms present in the immobilized culture, and particularly preferably, the microorganism of the species Paenibacillus macerans accounts for at least 1% of the total number of microorganisms present in the immobilized culture.
  • the microorganism of the species Paenibacillus macerans makes up at least 10% of the total number of microorganisms present in the immobilized culture, more preferably the microorganism of the species Paenibacillus macerans makes up at least 50% of the total number of microorganisms present in the immobilized culture, and is particularly preferred the microorganism of the species Paenibacillus macerans makes up at least 90% of the total number of microorganisms present in the immobilized culture.
  • at least one immobilized pure culture of a microorganism of the species Paenibacillus macerans is added.
  • Gel-forming polymers are preferably used. These have the advantage that bacteria can be taken up or stored within the gel structure. Preferably, those materials are used which dissolve slowly in water or are degraded, so that the release of the microorganism Paenibacillus macerans takes place over a longer period of time.
  • suitable polymers are polyaniline, polypyrrole, polyvinyl pyrolidone, polystyrene, polyvinyl chloride, polyvinyl alcohol, polyethylene, polypropylene,
  • Epoxy resins polyethyleneimines, polysaccharides such as agarose, alginate or cellulose,
  • Ethylcellulose methylcellulose, carboxymethylethylcellulose, cellulose acetates,
  • Polydialkyldimethylammonium chloride mixtures of polyacrylic acids and Polydiallyldimethylammoniumchlorid and mixtures thereof.
  • the polymeric material may also be prepared by conventional crosslinkers such as glutaraldehyde,
  • Urea / formaldehyde resins or tannin compounds are crosslinked.
  • Alginates as Immobilisate prove to be particularly advantageous because they do not have a negative impact on the activity of the microorganism Paenibacillus macerans and on the other hand they are slowly degraded by microorganisms. Due to the slow degradation of the alginate immobilizates, the trapped microorganisms of the species Paenibacillus macerans are gradually released.
  • the microorganisms are mixed with a polymer gel and then cured in a suitable hardener solution. For this they are first mixed with a gel solution and then dropped into a hardener solution of suitable height. The detailed procedures for immobilization are known to the person skilled in the art.
  • additional biomass is added to the fermentation reactor in a timely manner to the addition of the microorganisms described below.
  • Timely addition of additional biomass may occur within a period of 1 second to 3 days after addition of microorganisms, or it may occur simultaneously with the addition of microorganisms.
  • the space load in the fermentation reactor can be continuously increased or kept approximately constant by the continuous addition of new substrate, wherein the fermentation at all room loads, preferably at a space load of ⁇ 0.5 kg of organic dry matter per m 3 and day [kgoTS / m 3 d] , more preferably at a space load of ⁇ 4.0 kgoTS / m 3 d and particularly preferably at a room load of ⁇ 6.0 kgoTS / m 3 d can be performed, which corresponds to an increase in the space load to about double compared to the current state of the art.
  • the fermentation substrate used can in particular also have a high proportion of solid constituents.
  • a hydrolytically active, fermentative microorganism of the species Paenibacillus macerans By adding a hydrolytically active, fermentative microorganism of the species Paenibacillus macerans, these solid constituents are at least partially liquefied. Due to the liquefaction of the fermentation substrate due to the addition of the microorganism Paenibacillus macerans, a thickening of the fermenter material can be prevented and targeted counteracted. Another liquid entry into the fermentation substrate in the form of water or manure during fermentation can be avoided. Thus, there is another advantage in conserving the resource freshwater. Another advantage is the thus obtained obtaining the stirring and pumpability of the substrate. As a result, agitators and pumps are spared and significantly less energy is required for the stirring process.
  • microorganisms of the species Paenibacillus macerans can also be used for the liquefaction of biomass in alcoholic fermentation with the aim of producing biofuel.
  • the treatment of biomass takes place with constant mixing of the fermentation substrate. Due to the constant mixing of the fermentation substrate, the cultures of Paenibacillus macerans can be better distributed in the fermentation substrate. In the case of biogas production, moreover, the biogas produced can be better removed from the fermentation process.
  • the constant mixing of the fermentation substrate also leads to a uniform heat distribution in the fermentation reactor.
  • Measurements of the temperature in the fermentation reactor which were carried out at periodic intervals, but also continuously, showed that the fermentation substrate in a temperature range of 20 0 C to 80 0 C, preferably at about 35 0 C to 60 0 C, particularly preferably at 40 0 C is fermented efficiently to 50 0 C. These temperature ranges are therefore preferred in the context of the present invention.
  • the last stage of the fermentation process namely the formation of methane by methanogenic microorganisms, particularly efficient at elevated temperatures.
  • All embodiments of the present invention are not limited to one-step processes for the production of biogas.
  • the use of microorganisms of the species Paenibacillus macerans can also be carried out in two or more stages.
  • microorganisms of the species Paenibacillus macerans can be used in processes for the production of liquid biofuels.
  • fermentation substrate and a microorganism of the species Paenibacillus macerans are added continuously.
  • the continuous operation of a fermentation reactor should result in a stable microbial biocenosis to a continuous production of biogas, the exposure of the substrate addition to the fermentation should be reduced as a result of a process disturbance.
  • the microorganism of the species Paenibacillus macerans may be added at regular intervals to the fermentation substrate Addition of the microorganism of the species Paenibacillus macerans at regular intervals leads to an increase in the autismdzelliere and thus to an improved sequence of fermentative processes, such as hydrolysis with a simultaneous improved utilization of the fermentation substrate for fermentation.
  • the microorganism of the species Paenibacillus macerans is added in an amount to the
  • Fermentation substrate added so that after addition of the proportion of the microorganism of
  • Fermentation substrate present microorganisms.
  • Fermentation substrate present microorganisms.
  • a microorganism of the species Paenibacillus macerans is particularly preferably added in an amount to the fermentation substrate that makes up after addition of the content of the microorganism of the species Paenibacillus macerans between 10 "6% and 25% of the total number of present in the fermentation substrate microorganisms.
  • the microorganism of the species Paenibacillus macerans is added in an amount to the fermentation substrate that makes up after addition of the content of the microorganism of the species Paenibacillus macerans between 10 "4% and 10% of the total number present in the fermentation substrate microorganisms.
  • the microorganism of the species Paenibacillus macerans is added to the fermentation substrate in an amount such that, after addition, the proportion of the microorganism of the species Paenibacillus macerans is between 10 -3 % and 1% of the total number of microorganisms present in the fermentation substrate.
  • microorganisms of the species Paenibacillus macerans can be carried out at any point in the fermentation process; in particular, microorganisms of the species Paenibacillus macerans can also be used to inoculate fermentation substrate upon first use or restart of a fermenter.
  • Microorganisms of the species Paenibacillus macerans may be added in the form of a culture once or several times at regular or irregular intervals, but preferably weekly or monthly, more preferably daily or twice to five times per week in an appropriate concentration and amount. Suitable concentrations of microorganisms and added amounts have already been mentioned or are described in the working examples.
  • microorganisms of the species Paenibacillus macerans in case of disturbances in the fermentation process to stabilize the fermentation. Such disorders can be detected early by monitoring certain characteristic parameters of the fermentation. Characteristic parameters provide information about the quality of an ongoing fermentation process for the production of biogas.
  • Such characteristic parameters are not only the amount of biogas produced and the methane content of the biogas produced but also, for example, the hydrogen content of the biogas produced, the pH of the fermentation substrate, the redox potential of the Fermentation substrate, the carboxylic acid content of the fermentation substrate, the proportions of various carboxylic acids in the fermentation substrate, the hydrogen content of the fermentation substrate, the proportion of dry matter in the fermentation substrate, the proportion of organic dry matter in the fermentation substrate, the viscosity of the fermentation substrate and the volume loading of the fermentation.
  • the present invention also encompasses the use of a microorganism of the species Paenibacillus macerans for the treatment of biomass.
  • the present invention also includes the use of a microorganism of the species Paenibacillus macerans for the liquefaction of biomass.
  • the present invention also encompasses the use of a microorganism of the species Paenibacillus macerans for the fermentative production of biogas from biomass.
  • the present invention also encompasses the use of the liquefied biomass for biofuel production obtained by one of the described processes.
  • the liquefied biomass obtained by one of the processes described is preferably used for the production of bioethanol.
  • the present invention comprises the strain of the microorganism Paenibacillus macerans SBG2, as deposited under No. DSM 22569.
  • the microorganism Paenibacillus macerans SBG2 was deposited in a pure culture at the German Collection of Microorganisms and Cell Cultures GmbH in Braunschweig under the Budapest Treaty.
  • the name is Paenibacillus macerans SBG2 with the accession number DSM 22569.
  • the present invention comprises the strain of the microorganism Clostridium sartagoformum SBGIa, as deposited under No. DSM 22578.
  • the microorganism Clostridium sartagoformum SBGIa was deposited in a pure culture at the German Collection of Microorganisms and Cell Cultures GmbH in Braunschweig under the Budapest Treaty.
  • the name is Clostridium sartagoformum SBGIa with the accession number DSM 22578.
  • Bacteria of the species Paenibacillus macerans can be isolated from the fermentation substrate or fermentation residue of a fermenter with the aid of methods known to those skilled in the art.
  • a suitable substrate is introduced from a fermenter into a selection medium, cultured for a long time and finally isolated individual colonies of microorganisms from the selection medium.
  • microorganisms of the species Paenibacillus macerans can be selected on the basis of the DNA.
  • Bacteria Paenibacillus macerans SBG2 were isolated from the fermentation substrate of a post-fermenter. For this purpose, nitrogen and carbon dioxide were passed through a liquid selection medium, then Na 2 S was added to the selection medium and autoclaved (20 min. At 121 0 C). Then, the biomass obtained from the secondary digester was introduced into the selection medium and cultured for at least one week at a temperature status of at least 30 0 C. A sample obtained from the liquid selection medium was applied to a solid selection medium and subsequently the colonies of microorganisms grown on the solid selection medium were selected. After amplification of the obtained microbial DNA by PCR, a comparison with known DNA sequences could be performed.
  • the present invention also includes microorganisms having a nucleic acid having a nucleotide sequence containing a sequence region having more than 97.63% sequence identity with the nucleotide sequence of SEQ ID NO: 1. More preferably, the nucleotide sequence contains a sequence range greater than 97.64% or greater than 97.66% or greater than 97.68% or greater than 97.70% or greater than 97.75% or greater than 97.80%. or more than 97.90% sequence identity with the nucleotide sequence of SEQ ID NO: 1.
  • the nucleotide sequence contains a sequence region which has more than 98.0% sequence identity with the nucleotide sequence SEQ ID No. 1. More preferably, the nucleotide sequence contains a sequence range greater than 98.1% or greater than 98.2% or greater than 98.3% or greater than 98.4% or greater than 98.5% or greater than 98.6%. or more than 98.7% or greater than 98.8% or greater than 98.9% sequence identity with the nucleotide sequence SEQ ID NO: 1, and more preferably the nucleotide sequence contains a sequence region having greater than 99% sequence identity with the nucleotide sequence SEQ ID No. 1.
  • the microorganism has a nucleotide sequence containing a sequence region having more than 99.5% sequence identity with the nucleotide sequence SEQ ID NO: 1, and more preferably the nucleotide sequence contains a sequence region having greater than 99.8% sequence identity having the nucleotide sequence of SEQ ID NO: 1.
  • the nucleotide sequence contains a sequence region which corresponds to the nucleotide sequence SEQ ID No. 1.
  • SEQ ID NO: 1 may be compared to the starting nucleotide sequence at one or two positions or at three positions or at four positions or at five positions or at six positions or at seven positions or at eight positions or at nine positions or ten positions or eleven positions or twelve Positions or 13 positions or 14 positions or 15 positions or 16 positions or 17 positions or 18 positions or 19 positions or 20 positions or 21 positions or 22 positions or 23 positions or 24 positions or at 25 positions or at 26 positions or at 27 positions or at 28 positions or at 29 positions or at 30 positions or at 31 positions or at 32 positions or at 33 positions or at 34 positions or at 35 positions nucleotide mutations.
  • the meaning of the term "nucleotide mutation" is explained in the "Definitions" section of the present text.
  • the present invention also encompasses a culture of microorganisms suitable for use in a process for treating biomass, in particular a process for liquefying biomass and / or a process for the fermentative production of biogas from biomass, wherein a microorganism is present in the culture of microorganisms which has a nucleotide sequence containing a sequence region having at least 97.63% sequence identity with the nucleotide sequence of SEQ ID NO: 1, wherein the microorganism constitutes at least 10 "4 % of the total number of microorganisms present in the culture.
  • the nucleotide sequence contains a sequence region which has more than 99% sequence identity with the nucleotide sequence SEQ ID No. 1.
  • a microorganism in the culture of microorganisms suitable for use in a process for the treatment of biomass, in particular a process for the liquefaction of biomass and / or a process for the fermentative production of biogas from biomass, a microorganism is present which has a nucleotide sequence with a sequence region which has more than 99.5% sequence identity with the nucleotide sequence SEQ ID NO: 1.
  • the nucleotide sequence contains a sequence region which has more than 99.8% sequence identity with the nucleotide sequence SEQ ID No. 1.
  • a microorganism which has a nucleotide sequence is present in the culture of microorganisms suitable for use in a process for the treatment of biomass, in particular a process for liquefying biomass and / or a process for the fermentative production of biogas from biomass containing a sequence region corresponding to the nucleotide sequence of SEQ ID NO: 1.
  • the microorganism Paenibacillus macerans accounts for at least 10.sup.- 2 %, preferably at least 1% of the total number of microorganisms present in the culture.
  • the microorganism Paenibacillus macerans makes at least 10%, more preferably at least 25% of the total number in the Culture of existing microorganisms.
  • the microorganism Paenibacillus macerans accounts for at least 50%, in particular at least 90%, of the total number of microorganisms present in the culture.
  • it is a pure culture of microorganisms suitable for use in a method for the treatment of biomass, in particular a method for liquefying biomass and / or a method for fermentative production of biogas from biomass, which is a pure culture of the microorganism Paenibacillus macerans SBG2 as characterized above with respect to its nucleotide sequence.
  • it is an immobilized culture of microorganisms.
  • the present invention also encompasses an immobilized culture of microorganisms suitable for use in a method for the treatment of
  • Biomass in particular a process for the liquefaction of biomass and / or a process for the fermentative production of biogas from biomass, wherein in the immobilized culture of microorganisms a microorganism is present which has a nucleotide sequence containing a sequence region containing at least 97.63% Sequence identity with the nucleotide sequence SEQ ID NO: 1
  • No. 1 has.
  • a method for the treatment of biomass in particular a method for liquefying biomass and / or a method for the fermentative production of biogas from biomass suitable immobilized culture of microorganisms present a microorganism having a nucleotide sequence with a sequence region, more than 97.64% or more than 97.66% or more than 97.68% or more than 97.70% or more than 97.75% or more than 97.80% or more than 97.90% or more than 98.0% or more than 98.1% or more than 98.2% or more than 98.3% or more than 98.4% or more than 98.5% or more than 98.6% or greater than 98.7% or greater than 98.8% or greater than 98.9% sequence identity to the nucleotide sequence of SEQ ID NO: 1.
  • the nucleotide sequence contains a sequence region which has more than 99.0% sequence identity with the nucleotide sequence SEQ ID No. 1.
  • a microorganism which contains a nucleotide sequence is present in the immobilized culture of microorganisms suitable for use in a process for the treatment of biomass, in particular a process for the liquefaction of biomass and / or a process for the fermentative production of biogas from biomass
  • a process for the treatment of biomass in particular a process for the liquefaction of biomass and / or a process for the fermentative production of biogas from biomass
  • Very particularly preferably contains the Nucleotide sequence has a sequence region which has more than 99.8% sequence identity with the nucleotide sequence SEQ ID No. 1.
  • the invention is for use in a method for the treatment of biomass, in particular a method for liquefying biomass and / or a method for fermentative
  • Microorganisms include a microorganism having a nucleotide sequence containing a sequence region corresponding to the nucleotide sequence of SEQ ID NO: 1.
  • the present invention also encompasses the use of microorganisms as characterized above with respect to their nucleotide sequence in a method of treating biomass. These microorganisms are preferably used in one of the methods for the treatment of biomass explained in greater detail above.
  • the present invention also encompasses the use of microorganisms as characterized above with respect to their nucleotide sequence in a process for liquefying biomass. These microorganisms are preferably used in one of the above-explained methods for liquefying biomass.
  • the present invention also encompasses the use of microorganisms as characterized above with respect to their nucleotide sequence in a process for the fermentative production of biogas from biomass. These microorganisms are preferably used in one of the methods explained in more detail above for the fermentative production of biogas from biomass.
  • the present invention also encompasses the use of a culture of microorganisms as characterized above with respect to their nucleotide sequence in a method of treating biomass. These cultures of microorganisms are preferably used in one of the methods for the treatment of biomass explained in more detail above.
  • the present invention also encompasses the use of a culture of microorganisms as characterized above with respect to their nucleotide sequence in a process for liquefying biomass. These cultures of microorganisms are preferably used in one of the above-explained methods for liquefying biomass.
  • the present invention also encompasses the use of a culture of microorganisms as characterized above with respect to their nucleotide sequence in a process for the fermentative production of biogas from biomass. These cultures of microorganisms are preferably used in one of the methods explained in more detail above for the fermentative production of biogas from biomass.
  • Fig. 1 shows a substrate flow test for investigating the degree of liquefaction of fermentation substrate
  • Fig. 2 Results of a fermentation: Plotted is the gas yield as space-time yield (N l / l) and the space load of the fermenter against time;
  • Fig. 3 results of a further fermentation: Plotted is the gas yield as space-time yield (Nl / I) and the specific gas yield (Nm 3 A) of the fermenter against time; Ways to carry out the invention
  • Bacteria Paenibacillus macerans SBG2 were successfully isolated from the fermentation substrate of a post-fermenter. The deposit of the organism was carried out in a pure culture at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) according to the Budapest Treaty (Paenibacillus macerans SBG2 with the accession number DSM 22569).
  • Bacteria Clostridium sartagoformum SBGIa were also successfully isolated from the fermentation substrate of a post-fermenter. The deposit of the organism was carried out in a pure culture at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) according to the Budapest Treaty (Clostridium sartagoformum SBGIa with the accession number DSM 22578).
  • bacteria of the strain Clostridium sartagoformum SBGIa both in mixed culture and in pure culture properties according to the invention similar to those to bacteria of the strain Paenibacillus macerans SBG2, such. an increase in the volume of space, an increase in gas yield and a stabilization of the biogas process as well as the ability to liquefy biomass.
  • Bacteria of the strain Paenibacillus macerans SBG2 were successfully isolated from the fermentation substrate of a post-fermenter.
  • the microorganisms were isolated using a selection medium containing carboxymethylcellulose (CMC) as the sole carbon source.
  • CMC carboxymethylcellulose
  • Carboxymethylcellulose is very similar to the cellulose contained in fermentation substrates of biogas plants and, moreover, has an improved solubility in an aqueous medium due to the linking of the hydroxyl groups with carboxymethyl groups (-CH 2 -COOH-). That for selection Medium used by Paenibacillus macerans SBG2 (DSMZ medium 520 plus 1% CMC and 0.2% yeast extract) was gassed with N 2 for selection to be made under anaerobic conditions. Contained residual oxygen was reduced by means of 0.5 g / l Na 2 S.
  • the selection medium was then inoculated with the supernatant of material from a post fermenter (diluted 1: 2000). After one week of cultivation at 40 ° C., single rods were observed under microscopic analysis. Further selection of liquid cultures and isolation to pure cultures was achieved by smearing on anaerobic carboxymethylcellulose plates.
  • the cultivation was carried out under the specified conditions in a 1 m 3 fermenter, which was inoculated with 100 ml preculture. Since growth is quite fast with a doubling time of about 2 h, Paenibacillus macerans SBG2 is particularly suitable for biotechnological application.
  • Smaller amounts of bacteria eg for addition to fermenter types 1, 2 and 4) were cultured in 500 ml to 1 l scale. Cultivation for the addition to a fermentation process took place over a period of 1 to 2 days. Cell densities in the range of 10 8 to 10 10 cells per ml of culture medium were achieved. The bacterial cells were harvested by centrifugation and taken in the smallest possible volume of fresh medium before they were used in the fermentation process. For interim storage, the cells were frozen.
  • Microorganisms of the species Clostridium sartagoformum and Clostridium sporosphaeroides e.g. for the use of mixed cultures with approximately equal proportions of the 3 mentioned microorganisms could be cultured under the same conditions.
  • the cell material of the grown sporadic colonies was used for amplification of the microbial DNA by the Colony PCR method according to a standard program.
  • the gene for the 16S rRNA was amplified from the cell DNA by PCR.
  • the primers with the sequences GRGTTTGATCCTGGCTCAG and ACGGHTACCTTGTTACGACTT were used (indicated in 5 " ⁇ 3 " direction, H is C, T or A).
  • the pieces of DNA obtained as PCR products were then cloned into a cloning vector (ligation with the QIAGEN PCR cloning kit from QIAGEN / Hilden using the vector p-drive), transformed into E.
  • coli (according to QIAGEN PCR cloning - Handbook) and examined by colony PCR.
  • the obtained colony PCR products were subjected to restriction fragment length polymorphism analysis (RFLP) to select suitable clones. From the respective clones, the corresponding plasmid DNA was isolated and sequenced by the chain termination method (Sanger et al., 1977).
  • RFLP restriction fragment length polymorphism analysis
  • the 16S rDNA or its transformation into the corresponding 16S rRNA could be phylogenetically analyzed with the program package ARB (Ludwig et al., Nucleic Acids Research., 2004, 32, 1363-1371) and classified as an organism of the species Paenibacillus macerans become.
  • An analysis of the cloned 16S rDNA or the corresponding 16S rRNA sequence by means of BLAST program (basic local alignment search tool) of the database www.ncbi.nlm.nih.gov the clone Paenibacillus sp. H 10-05 determined as the next relative.
  • Maintaining a liquid-pulpy consistency in a wet fermentation process is essential to ensure a smooth and cost-effective process of the technical process.
  • continuous fermentation also leads to an increase in the fermenter content, which also has a negative effect on the biogas production process.
  • the effect that the addition of microorganisms has on the consistency of the fermentation substrate was determined by a test in which the flow behavior of substrate samples was measured on an inclined plane (substrate flow test).
  • Starting material for the flow test was material from a biogas plant with a dry matter content of about 8-12%.
  • Each 500 ml of material from a fermenter was filled into 1 I Schott bottles and incubated for 1 day at 40 0 C.
  • each of the bacterial cell mass from a 500 ml preculture (equivalent to approximately 4 x 10 11 cells) of Paenibacillus macerans SBG2, Clostridium sartagoformum SBGIa, Clostridium sporosphaeroides SBG3 or a mixture of the three bacteria was resuspended in equal proportions in 1 ml of medium, and added for further Incubated for 5 days at 40 ° C. with shaking.
  • FIG. 1 shows the following traces in FIG. 1: Lane 1: control without addition of microorganisms (comparative example); Lane 2: Clostridium sartagoformum SBGIa (comparative example); Lane 3: Clostridium sporosphaeroides SBG3 (comparative example); Lane 4: Paenibacillus macerans SBG2; Lane 5: Mixture of Clostridium sartagoformum SBGI a, Clostridium sporosphaeroides SBG3 and Paenibacillus macerans SBG2 in equal parts.
  • FIG. 1A shows the traces directly after the application of the samples, FIG. 1B after 2 min. and Figure 1 C after 18 min.
  • Clostridium sporosphaeroides SBG3 or Paenibacillus macerans SBG2 or Clostridium sarta ⁇ oformum SBGI a The proportion of bacteria of the species Clostridium sporosphaeroides SBG3 or Paenibacillus macerans SBG2 or Clostridium sartagoformum SBGI a was determined by whole cell hybridization according to the method described in Amann et al., (1995, Microbiol Rev 59, 143-169) Probe for fishing was used for Clostridium sporosphaeroides SBG3 a labeled oligonucleotide with the sequence Cy3-CCACAGCTCTCACGCCCG (indicated in 5 " ⁇ 3 " direction), for Paenibacillus macerans SBG2 a labeled oligonucleotide having the sequence Cy3- GCAACCCGAACTGAGACC (indicated in
  • Fermenter type 1 Lying plug fermenter rectangular, volume 150 1, subdivision of the fermenter compartment by retracted wall with small-area passage for the substrate flow, division of the fermenter space in VA directly after substrate addition nozzle and 3 A to pinhole, 2-stage system.
  • Fermenter Type 2 Lying plug fermenter rectangular, volume 150 I as 1st stage plus totally mixed round fermenter, volume 200 I as 2nd stage; Total volume 350 I. 2-stage plant with recirculation between round fermentor and plug-flow fermenter.
  • Fermenter type 3 horizontal cylindrical plug-flow fermenter, volume 30 m 3 , single-stage plant. Distribution of microorganisms within the fermenter - "Tracer test" Since the mixing of the viscous substrate and thus also other additives in a fermenter takes some time and also depends on the stirring technique used, a test was established with which the period to a uniform In this "tracer test", instead of microorganisms, powdered LiCI was added at a concentration of 10 mg lithium / kg starting substrate (double the amount for fermenter type 2) at the site of substrate addition.
  • Stable operation was achieved at a volume loading of 6 to 7 kgoTS / m 3 d by adding about 1 x 10 13 cells per m 3 and week. In doing so, nothing was changed in the usual process parameters, such as temperature (40 ° C.) and pH value (6-9), or buffer capacity.
  • gas yield and volume load In addition to gas yield and volume load, a number of other characteristic parameters of the fermentation process such as dry matter content of the fermenter content, pH, acid concentration (eg acetic acid, propionic acid, butyric acid, valeric acid, acetic acid equivalent), temperature, specific gas yield, composition of the biogas, viscosity, conductivity, redox potential and concentration measured on nutrients and trace elements.
  • acid concentration eg acetic acid, propionic acid, butyric acid, valeric acid, acetic acid equivalent
  • This pilot plant is a large test facility on a pilot plant scale with a volume of 30 m 3 , which is constructed as a horizontal cylindrically shaped plug-flow fermenter. It is operated as a 1-stage system.
  • FIG. 2 shows measurement results of various characteristic parameters during a fermentation process in a type 3 experimental fermenter with and without the addition of microorganisms of the strain Paenibacillus macerans SBG2.
  • the curve provided with the reference numeral 10 shows the time course the volume load of the fermenter in kilograms of organic dry matter per cubic meter per day (kgoTS / m 3 d), the curve indicated by the reference numeral 20 the time course of the measured space-time yield (Nl / I), the curve indicated by the reference numeral 30, the time course of measured space-time yield, averaged over 6 days.
  • the reference numeral 40 the time course of the theoretical gas production (Nl / I) is marked.
  • the reference numeral 50 marks the symbols each representing the one-time addition of microorganisms of the strain Paenibacillus macerans SBG2.
  • the biogas plant was operated from day 70 to day 102 without external addition of microorganisms at an average volume load of about 4 kgoTS / m 3 d and provided in a stable fermentation process space-time yields averaging 2.8 Nl / I, slightly above the expected theoretical Gas yield lay.
  • 20 l of water were added each day instead of a bacterial culture.
  • 20 l of a pure culture of Paenibacillus macerans SBG2 (bacterial concentration about 2 ⁇ 10 9 cells / ml) were added daily.
  • FIG. 3 shows measurement results of various characteristic parameters during a fermentation process in a type 3 experimental fermenter with and without the addition of microorganisms of the species Paenibacillus macerans SBG2.
  • the curve provided with the reference numeral 10 shows the time course of measured specific gas yield (Nm 3 / t), the curve indicated by the reference numeral 20, the time course of the measured specific gas yield (Nm 3 A), averaged over 6 days.
  • the curve provided with the reference numeral 30 shows the time course of the measured space-time yield (Nl / I), the curve indicated by the reference numeral 40, the time course of the measured space-time yield (Nl / I), averaged over 6 days.
  • the arrow marks the beginning of the addition of microorganisms of the strain Paenibacillus macerans SBG2 from day 102.
  • the space-time yield (NI / I) in the Type 3 fermenter rose very sharply after the addition of microorganisms of the strain Paenibacillus macerans SBG2 because the volume load could be greatly increased. With increased substrate supply thus increased in a still stable fermentation process, the gas yield achieved per fermenter volume. It can be seen from the experimental results shown in FIG. 3 that the specific gas yield (Nm 3 A) also increased significantly after the addition of microorganisms of the strain Paenibacillus macerans SBG 2 (see curves with the reference numbers 10 and 20).
  • the average specific gas yield was about 720 Nm 3 / t substrate, after addition of microorganisms of the strain Paenibacillus macerans SBG2 about 780 Nm 3 / t substrate.
  • the measured variable "specific gas yield" is not normalized to the fermentation volume, but to the amount of substrate used, ie an increase in the specific gas yield means a higher energy efficiency in the substrate conversion, but an economic advantage for the user is an increase in the space-time yield as well an increase in the specific gas yield.
  • the volume load in fermentations could be increased to a maximum value of about 8 kgoTS / m 3 d.
  • the space load of the plant could be further increased as a result of the addition of Paenibacillus macerans SBG2.
  • the percentage content of dry substance or organic dry matter remained almost constant. This observation suggests that no accumulation of non-fermented organic dry substance occurs during the fermentation of the fermentation substrate.
  • the addition of pure cultures of Paenibacillus macerans SBG2 thus contributes to a continuous conversion of the dry matter contained in the fermentation substrate, which in turn leads to a continuous fermentation by the accumulation of dry matter is reduced.
  • pure cultures of Paenibacillus macerans but also mixed cultures with a proportion of Paenibacillus macerans were used.
  • mixed cultures of two or three kinds of microorganisms selected from the group consisting of Paenibacillus macerans, Clostridium sartagoformum and Clostridium sporosphaeroides can be added.
  • the addition of the hydrolytically active, fermentative microorganism Paenibacillus macerans SBG2 has a positive effect on the hydrolysis of organic dry matter.
  • the volume load of a fermenter can be increased from about 4 to 5 kgoTS / m 3 d to about 6 to 8 kgoTS / m 3 d under otherwise identical conditions, without even instability of the fermentation process would suggest.
  • Parallel to the increased space load the amount of biogas produced is significantly increased.
  • the specific yield of biogas increases, since significantly more of the organic dry matter is degraded than in the absence of addition of microorganisms of the species Paenibacillus macerans.
  • the use of microorganisms of the species Paenibacillus macerans leads to a dramatic improvement in the efficiency and efficiency of biogas plants.

Abstract

L'invention concerne un procédé pour traiter de la biomasse. Selon ce procédé, un micro-organisme du type Paenibacillus macerans est ajouté à la biomasse.
PCT/DE2009/075036 2008-12-23 2009-07-07 Paenibacillus macerans pour traiter de la biomasse WO2010072220A1 (fr)

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CN102557766A (zh) * 2012-02-21 2012-07-11 浙江大学 浸麻类芽孢杆菌菌株在溶解土壤或富营养化水体中磷源的应用
CN102557766B (zh) * 2012-02-21 2013-12-11 浙江大学 浸麻类芽孢杆菌菌株在溶解土壤或富营养化水体中磷源的应用
WO2014013509A1 (fr) * 2012-07-19 2014-01-23 Council Of Scientific And Industrial Research Biotransformation combinée de biomasse lignocellulosique pour la production d'acide l-lactique
EP4043544A1 (fr) * 2021-01-20 2022-08-17 WAS Wirtschaftsagentur Martin Schroeder GmbH Procédé et dispositif de valorisation de matières organiques

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