WO2013163703A1 - Procédé intégré de production de formulations enzymatiques à partir de déchets agro-industriels et production de biocombustibles - Google Patents

Procédé intégré de production de formulations enzymatiques à partir de déchets agro-industriels et production de biocombustibles Download PDF

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WO2013163703A1
WO2013163703A1 PCT/BR2012/000123 BR2012000123W WO2013163703A1 WO 2013163703 A1 WO2013163703 A1 WO 2013163703A1 BR 2012000123 W BR2012000123 W BR 2012000123W WO 2013163703 A1 WO2013163703 A1 WO 2013163703A1
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Prior art keywords
enzymatic
production
enzyme
formulations
fermentation
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PCT/BR2012/000123
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English (en)
Portuguese (pt)
Inventor
Aline Machado De Castro
Mariana MARTINS PEREIRA TEIXEIRA
Daniele FERNANDES CARVALHO
Carolina DA COSTA LÁZARO
Denise Maria GUIMARÃES FREIRE
Leda Dos Reis Castilho
Lidia Maria Melo Santa Anna
Danuza Nogueira Moyses
Absai DA CONCEIÇÃO GOMES
Claudia Julia Groposo Silveira
Vinicius Azevedo DE ARAUJO
Bernardo ALVES CINELLI
Jimmy Andrés LÓPEZ JIMÉNEZ
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Petróleo Brasileiro S.A. - Petrobras
Universidade Federal Do Rio De Janeiro - Ufrj
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Priority to BR112013005916-8A priority Critical patent/BR112013005916B1/pt
Priority to PCT/BR2012/000123 priority patent/WO2013163703A1/fr
Publication of WO2013163703A1 publication Critical patent/WO2013163703A1/fr

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Classifications

    • 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
    • C10L1/00—Liquid carbonaceous fuels
    • C10L1/02—Liquid carbonaceous fuels essentially based on components consisting of carbon, hydrogen, and oxygen only
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/10—Process efficiency

Definitions

  • Such formulations find application for converting starch-rich biomasses, as well as those containing lignocellulose, into sugar-rich streams that can then be fermented to produce biofuels, especially ethanol, and "green" chemicals such as organic acids, biopolymers, antibiotics and polyols.
  • the process of obtaining such formulations is conducted in an integrated manner, aiming to reduce the energy expenditure involved in the processes of this nature currently in use, since the formulations obtained have all the enzymes necessary for the granular starch hydrolysis process and conversion to ethanol and others. Chemicals.
  • ground corn kernels are subjected to a cooking step in which a suspension containing about 30 to 35% solid material is heated and kept at a temperature of at least 100 ° C for about 20 minutes in the presence of calcium hydroxide and ammonia - gelatinization step.
  • the solubilization of the starch contained in the material makes it more exposed to the action of the enzymes that are used in the later stages, when starch hydrolysis occurs.
  • an endoamilolytic concentrate consisting of alpha-amylase enzymes produced by bacteria is employed. The reaction is conducted at a temperature in the range of 80 ° C to 90 ° C for 30 to 120 minutes.
  • Starch polysaccharides (molecules with only linear bonds such as amylose and with linear and branched bonds such as amylopectin) are attacked and release dextrins (medium polymerized glucose oligosaccharides).
  • Sulfuric acid solution is then added to the medium by changing the pH of the medium from 6.0 to 4.5 and introducing another group of enzymes, glucoamylases, generally produced by filamentous fungi, which have the function of catalyze the hydrolysis of dextrins, finally releasing glucose.
  • the reaction is conducted at a temperature in the range of 60 ° C to 70 ° C for at least 5 hours. A high glucose syrup is obtained.
  • the glucose-rich syrup is then fed to a fermenter, where it is converted to ethanol by microbiological action.
  • the main co-product containing protein, fiber and unconverted starch (Distillers Dried Grains with Solubles - DDGS) is used as animal feed.
  • the energy balance of the process is considered unfavorable (McAloon and collaborators - NREL Techincal Report 580-28893, 2000), since the The energy balance for ethanol production from sugarcane in Brazil is 8.9 (Moreira - Energy Sust. Develop, 4:43, 2000). More recent study finds very promising use of a technology for converting starch materials whose hydrolysis process is conducted at a low temperature - below the starch solubilization temperature - called "cold hydrolysis", which provides a reduction of up to 17%. of energy demand (Mueller - An analysis of the projected energy use of future dry mill corn ethanol plants 2010/2030).
  • the starch in its granular form, is subjected to a previous saccharification step in order to become more susceptible to hydrolysis.
  • the reaction is conducted at 57 ° C for 2 hours at acid pH and in the presence of enzyme preparations containing endoamylases and accessory enzymes such as proteases and cellulases, which aid in the degradation of starch-associated proteins and polysaccharides.
  • the suspension further containing granular starch and some free glucose content, is conveyed to a fermenter and receives the addition of an enzyme preparation containing endoamylases and exoamylases, capable of hydrolyzing the starch in its native form, and a yeast such that saccharification of dextrins and fermentation of released sugars occur simultaneously.
  • US2009 / 0017511 describes a process for producing ethanol from granular starch comprising a first step in which a slurry of water and solubilized starch is obtained which is pre-treated at elevated temperature in the presence of a acid alpha-amylase and a glycoamylase at a temperature between 0 ° C and 20 ° C below the initial gelatinization temperature for a period of 5 minutes to 12 hours.
  • the sludge is fermented in the presence of an alpha-amylase, a glycoamylase and a yeast at a temperature in the range of 10 ° C to 35 ° C for 20 to 250 hours to produce alcohol, and is preferable.
  • a phytase for the reaction to occur in the presence of a phytase.
  • various raw materials can be used, such as corn, wheat, oats, rye, barley, cassava, sorghum, among others.
  • the enzymes used in the process are commercial enzymes obtained from strains of bacteria and fungi that have undergone some kind of genetic modification.
  • US6667066 teaches the production of a multi-enzyme product with glycoamylase, protease and xylanase obtained by fermentation of wheat bran with an Aspergillus niger strain. units per gram of dry material. The product has application to produce ethanol or monogastric animal feed.
  • US2008 / 0003344 describes a process for producing ethanol from the hydrolysis of starch contained in ground vegetable waste using a combination of enzymes.
  • enzymes such as phytase, glucoamylase and alpha-amylases of microbiological origin, aiming to achieve hydrolysis at temperatures below starch gelatinization.
  • beta-glucosity in combination with other enzymes.
  • the final toxin free residue can be used as animal feed.
  • US201 1/0097446 describes a method for producing high ethanol contents from starch-containing plant materials (corn, wheat, oats, sorghum, among others).
  • the method consists of milling the plant material to achieve a particle size of between 0.1 and 0.5 mm, subjecting the material to saccharification without starch cooking in the presence of specific enzymes, fermenting the incubated starch in the presence of a yeast. and recover the ethanol from the fermented medium. During fermentation the temperature is varied.
  • the method allows to recover at least 18% v / v ethanol.
  • the process makes use of commercial enzymes with specific activities for starch hydrolysis.
  • WO20 / 0 7093 teaches the use of alpha-amylase mixtures in the liquefaction and saccharification of starch for ethanol production.
  • the mixture employs enzymes used in the ratio of 1 unit of low pH thermostable alpha-amylase for every 5 Modified Wohlgemuth Units (MWU) of Bacillus licheniforms.
  • MNU Modified Wohlgemuth Units
  • the present invention is concerned with obtaining enzyme formulations containing enzymes produced by solid state fermentation, a low cost process, so that said formulations have characteristics that allow application to different especially aimed at improving ethanol production from different sources of properly balanced biomass.
  • the presence of free amino acids easily assimilated by cells is an important differential of the invention.
  • the invention relates to the production of enzyme formulations produced from agro-industrial waste, in particular waste from the vegetable oil and brewery industries. More particularly, it is also intended to prepare well-balanced microorganism culture medium formulations for simultaneous production via solid state fermentation of all enzyme groups required for granular starch hydrolysis.
  • the obtained enzymatic formulations are intended for the conversion of starch-rich biomasses or those containing lignocellulose, with a view to generating streams rich in fermentable sugars for later production of biofuels, particularly ethanol, and "green chemicals" (organic acids, biopolymers, antibiotics and polyols, among others).
  • the invention aims at the application of such enzyme formulations containing amylases (alpha-amylases, glucoamylases, isoamylases), cellulases (endoglucanases, beta-glucosidases, cellobiohydrolases), hemicellulases (xylanases, beta-xylosidases) and proteases (endopeptidases), exopeptidases, exopeptidases, exopeptidases, exopeptidases, exopeptidases.
  • FIGURES Figure 1 shows a block diagram of the process. of the present invention.
  • Figure 2 shows profiles of glucose consumption (hollow squares) and ethanol production (filled squares) obtained from the babassu flour fermentation process, as well as glycerol production (filled triangles). The concentrations of the molecules are expressed in g / l.
  • the integrated process of the present invention aims to establish balanced formulations of solid culture medium for microorganisms in order to simultaneously produce all enzyme groups necessary for granular starch hydrolysis.
  • agroindustrial residues are used for the production of these enzyme formulations, particularly residues originating from the vegetable oil and brewery industry, so that their production cost becomes greatly reduced.
  • the culture media preparation process should initially consider the characteristics of the different agroindustrial residues to be used and then acclimate the microorganisms to produce the desired enzymes.
  • the integrated process comprises the following steps:
  • agroindustrial residue (10) with particle size in the range of 0.05 - 2.00mm, a first stream of water (15), to adjust and maintain the moisture content in the range of 40 - 80%, and previously propagated filamentous fungi (20) with an initial concentration of 10 3 - 10 7 / ml, selected from fungal strains, for example Aspergillus, to form a solid matrix (30); maintain fermentation (F1) in solid state (FES) for 12 - 168h at a temperature in the range 20 ° C - 55 ° C;
  • F1 solid state
  • this step may not be required and the enzyme extract (40) may be sent directly to the enzymatic hydrolysis and submerged fermentation reactor (F3);
  • M raw material formed from residue and / or starch-rich agroindustrial material (10) such as babassu flour, corn kernels, wheat kernels, sorghum kernels, castor cake and formulation enzyme (45) to a second reactor (200), to undergo enzymatic hydrolysis and submerged fermentation (F3), in the presence of the enzyme formulation (45) obtained, using microbial cells (60) suitable for the production of the desired product (70), said microbial cells selected from bacteria, actinomycetes, yeast and filamentous fungi.
  • microbial cells selected from bacteria, actinomycetes, yeast and filamentous fungi.
  • enzyme production is achieved via solid state fermentation.
  • Fungal strains are maintained for 3 to 7 days of static cultivation in solid culture medium containing polysaccharides as carbon source, aiming at the acclimatization of cells to produce enzymes capable of degrading such molecules.
  • the spores at a rate of 10 3 to 10 7 / ml, are inoculated in liquid medium for cell propagation at a temperature in the range of 20 ° C to 40 ° C under orbital agitation in the range of 100 to 300 ° C. rpm for a period of 10 to 48 hours.
  • babassu pie obtained from pressing babassu almonds
  • babassu flour obtained from pressing babassu mesocarp, castor pie
  • canola pie obtained from pressing babassu mesocarp
  • sunflower pie canola bagasse
  • starch corn sugarcane molasses
  • soybean molasses corn (corn maceration residual water)
  • soybean biodiesel crude glycerin sugarcane juice
  • Agroindustrial residues are mixed in different proportions and combinations in order to induce, in a customized way, the synthesis of enzymatic formulations, with balanced activity between the enzymes of interest, for subsequent hydrolysis of different biomasses and release of fermentable sugars to be used. in the process of obtaining the final product.
  • Balanced culture medium formulations are studied, combining the simultaneous supplementation of solid and liquid materials with babassu cake to customize its composition.
  • the microorganisms used are selected from Aspergillus fungi strains, particularly A. awamori, A. wentii and A. oryzae.
  • the enzyme extraction step is promoted. This extraction is done with distilled water at a ratio of 5: 1 to 20: 1 in relation to the initial cake mass, followed by maceration of the fermentation residue.
  • the material is then placed under agitation in the range of 100 to 300 rpm for a period of 10 to 120 min at a temperature in the range of 20 ° C to 55 ° C, preferably at 37 ° C. Then the material is centrifuged at 11,000 g and lyophilized. An enzymatic extract is obtained as a brown soluble powder which can be vacuum packed and stored at -20 ° C.
  • amylases alpha-amylases, glucoamylases, isoamylases
  • cellulases endoglucanases, beta-glucosidases
  • hemicellulases xylanases
  • proteases endopeptidases, exopeptidases
  • invertases contributes to the rapid assimilation of sugarcane juice sucrose
  • Beta-galactosidase allows dairy industry residues such as whey to be added to starch and lignocellulosic materials in the same reaction environment.
  • endoglucanase and beta-glucosidase indicates that the enzymatic preparation of the invention can act for the complete solubilization of cellulosic fibers until the release of glucose, its fermentable unit.
  • Hydrolysis testing experiments are performed at different temperatures, ranging from 30 ° C to 60 ° C.
  • Babassu flour is added in concentrations ranging from 3.33% to 25% (w / w), and hydrolysis is performed in small stirred reactors.
  • Sodium azide (0.2 g / L) is added in all hydrolysis experiments to avoid contamination.
  • VHG Very High Gravity
  • JP1 yeast cells (yeast strain from the Japungu -Santa Rita -Paraiba plant) are kept in Petri dishes on YPD agar medium (10g / L yeast extract, 20g / L peptone, 20g / L glucose, 20 g / L agar). After 48 or 72 hours in this medium at 30 ° C, the cells are aseptically transferred to a plastic tube with sterile distilled water, and thus the cell mass concentration calculated. With the cellular concentration, the YPD medium (10 g / L yeast extract, 20 g / L peptone, 20 g / L glucose) is inoculated and the pH adjusted to 5 with sulfuric acid.
  • the grown cells are inoculated into solid media, suitably formulated from agro-industrial materials.
  • babassu cake containing 62% of initial moisture with supplements the enzymes are generated, and these are extracted by adding water and stirring the system.
  • babassu flour By adding babassu flour to the enzyme extract at 50 ° C for 4 hours, fermentable sugars and free amino acids are released into the reaction medium.
  • This fermentation medium is previously propagated cells are added.
  • the inoculum concentration is 10% relative to the volume of the fermentation medium (corresponding to 0.5 g / l cells).
  • the submerged fermentation (FS) step is conducted at 32 ° C and under constant agitation for up to 72 hours.
  • the various enzymes present in the enzyme extract obtained act together in the hydrolysis of starch in granular form, releasing glucose. Simultaneously, Saccharomyces cerevisiae yeast cells consume glucose producing ethanol. Furthermore, in the enzymatic extract obtained according to the present invention, there are accessory enzymes such as xylanases, proteases and cellulases, which represent considerable technical and economic advantages.
  • the extraction step begins, obtaining the enzymes and free amino acids and peptides.
  • a culture medium suitable for various fermentative processes is formulated, containing all the basic nutritional sources necessary for the growth of different microorganisms.
  • the enzymes are used directly in liquid form, or the liquid extract is lyophilized for later redissolution, or the fermented solid mass undergoes proper drying and conditioning treatment.
  • the lyophilized enzyme extract is solubilized in water providing an exoamylolytic activity of 20 U / mL.
  • a suspension with 19% solids from babassu flour is prepared.
  • the pH is adjusted to 4.8 with sulfuric acid and the medium is thermostated at 50 ° C. for 4 hours, and then kept at 32 ° C for up to 72 hours. During incubation, the suspension is stirred continuously.
  • the concentration of assimilable nitrogen present in the hydrolyzed medium can meet all nutritional needs of yeast cells, confirming that it can be used as a fermentation medium for subsequent bioprocesses aimed at producing ethanol and other chemicals. It is further found that a FAN concentration between 300 and 400 mg / L is consumed during such subsequent fermentations.
  • the enzymes used in the process of the invention at low temperature act on the starch in granular form, thus eliminating the need for high energy demand for starch processing, providing higher glucose production for conversion to ethanol and other bioproducts. of higher added value.
  • Example 3 Use of low temperature enzyme extract
  • a second experiment is performed under the same conditions as described in the previous Example, but changing the temperature.
  • Babassu flour is also used, varying only the temperature, which is kept from the beginning at 32 ° C, lasting up to 24 hours.
  • Example 2 the process is conducted under the same conditions as described in Example 2, but using as a source of starch corn flakes smaller than 0.6 mm (28 mesh Tyler).
  • Babassu flour content in the range of 5% to 20% (by mass, relative to the total mass of the system), are added after 2, 4 and 24 hours of saccharification, totaling an addition between 10 and 50% of the solids contained in the system. initial step.
  • Example 2 using the enzyme formulation produced by solid state fermentation with Aspergillus awamori IOC-3914 in the presence of babassu flour, with 10% of the initial solids content added after 2, 4 and 24 h saccharification, totaling addition 30% of the solids contained at the beginning
  • babassu flour aiming at ethanol production
  • a suspension with 19% babassu flour is prepared.
  • the pH is adjusted to 4.8 with 7M sulfuric acid solution when required.
  • the medium is placed at 50 ° C for 4 hours, and then kept at 32 ° C for up to 72 hours.
  • Fermentation is conducted without any nitrogen source addition as the material from the biomasses themselves used to produce the enzyme formulation is hydrolyzed by the enzymes themselves and contains all the enzymes necessary for the final fermentation, including amylases (alpha-amylases, glucoamylases). , isoamylases), cellulases (endoglucanases, beta-glycosidases), hemicellulases (xylanases) and proteases (endopeptidases, exopeptidases).
  • the graph shown in Figure 2 shows the result of the kinetic profile of ethanol production during fermentation.
  • VHG Very High Gravity
  • Table 7 below shows the result of fermentation experiments with Saccharomyces cerevisae strain JP1.
  • the enzyme formulations of the invention besides being produced at low cost, have the advantage of having application in the simultaneous conversion of oligosaccharides and polysaccharides of different materials aiming to obtain biofuels (ethanol, butanol) and other green chemicals (acids biopolymers, antibiotics and polyols, among others), enabling an integrated processing of industrial plants and, consequently, reducing operating costs.
  • a suspension with 19% babassu flour is prepared.
  • the pH is adjusted to 4.8 with sulfuric acid when required.
  • the middle is. It is placed at 50 ° C for 24 hours and then the residual solid is separated by filtration and / or centrifugation. The liquid from this separation is then added to the yeast cells and the fermentation process is conducted for up to 48 hours at 32 ° C.
  • Table 8 below shows the result of fermentation experiments with Saccharomyces cerevisae strain JP1.
  • Table 9 below shows the result of fermentation experiments with Saccharomyces cerevisae strain JP1.

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Abstract

L'invention concerne l'obtention et l'utilisation de formulations enzymatiques en vue de l'hydrolyse d'amidon granulaire à partir de déchets agro-industriels. Ces formulations sont obtenues au moyen d'un procédé à faible coût et, après leur production, trouvent une application dans le co-traitement de matières premières avec différentes compositions, en vue de la conversion de biomasses riches en amidon ainsi que de celles contenant de la lignocellulose, dans des flux riches en sucres pouvant être ultérieurement fermentés pour la production de biocombustibles, notamment d'éthanol, et de produits chimiques "verts" - acides organiques, biopolymères, antibiotiques et polyols. Le procédé d'obtention de ces formulations est réalisé de manière intégrée, en vue de réduire la dépense énergétique qu'impliquent les procédés de cette nature.
PCT/BR2012/000123 2012-05-03 2012-05-03 Procédé intégré de production de formulations enzymatiques à partir de déchets agro-industriels et production de biocombustibles WO2013163703A1 (fr)

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BR112013005916-8A BR112013005916B1 (pt) 2012-05-03 2012-05-03 processo integrado para produção de biocombustíveis
PCT/BR2012/000123 WO2013163703A1 (fr) 2012-05-03 2012-05-03 Procédé intégré de production de formulations enzymatiques à partir de déchets agro-industriels et production de biocombustibles

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PCT/BR2012/000123 WO2013163703A1 (fr) 2012-05-03 2012-05-03 Procédé intégré de production de formulations enzymatiques à partir de déchets agro-industriels et production de biocombustibles

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Cited By (1)

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WO2016097638A1 (fr) 2014-12-18 2016-06-23 Veolia Proprete Biostimulation in-situ de l'hydrolyse de la matière organique pour optimiser sa valorisation énergétique

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016097638A1 (fr) 2014-12-18 2016-06-23 Veolia Proprete Biostimulation in-situ de l'hydrolyse de la matière organique pour optimiser sa valorisation énergétique
FR3030573A1 (fr) * 2014-12-18 2016-06-24 Veolia Proprete Biostimulation in-situ de l'hydrolyse de la matiere organique pour optimiser sa valorisation energetique
US10457967B2 (en) 2014-12-18 2019-10-29 Veolia Environment—Ve In-situ biostimulation of the hydrolysis of organic matter for optimizing the energy recovery therefrom

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