WO2019170788A1 - Procédé de fermentation anaérobie amélioré - Google Patents

Procédé de fermentation anaérobie amélioré Download PDF

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Publication number
WO2019170788A1
WO2019170788A1 PCT/EP2019/055629 EP2019055629W WO2019170788A1 WO 2019170788 A1 WO2019170788 A1 WO 2019170788A1 EP 2019055629 W EP2019055629 W EP 2019055629W WO 2019170788 A1 WO2019170788 A1 WO 2019170788A1
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lactic acid
antimicrobial compound
fermentation
process according
reactor
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PCT/EP2019/055629
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English (en)
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Ilco Adrianus Lambertus Antonius Boogers
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Dsm Ip Assets B.V.
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Publication of WO2019170788A1 publication Critical patent/WO2019170788A1/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
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/04—Bioreactors or fermenters specially adapted for specific uses for producing gas, e.g. biogas
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/12—Bioreactors or fermenters specially adapted for specific uses for producing fuels or solvents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • 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
    • 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 process for producing a valuable compound through anaerobic fermentation, to a set-up for running an anaerobic fermentation process suitable for producing a valuable compound, and to the use of a lactic acid sensitive biosensor in a set up for running an anaerobic fermentation process suitable for producing a valuable compound to reduce the dosage of antimicrobial compound.
  • yeast Saccharomyces cerevisiae is the established microbial cell factory for conversion of starch and sucrose derived hexose units to ethanol, as it combines a high ethanol yield and productivity with robustness under process conditions. Efforts in yeast strain improvement and process optimization of corn-starch and cane sugar based bioethanol production have further improved product yields and productivity.
  • yeast strains capable of efficiently fermenting the pentose sugars xylose and arabinose, thus paving the way for yeast based ‘second-generation’ bioethanol production from lignocellulosic hydrolysates.
  • biogas via the anaerobic digestion of organic material is a rapidly growing source of renewable energy.
  • the process is complex; a combined action of several biotechnological processes determines the stability, efficiency and yield of the biogas produced.
  • An optimal process design is still under active research done at laboratory and pilot plants. Substrates like grass, manure or sludge can be used as feed for the biogas production due to their high yield potential
  • Bacterial contamination represents a persistent problem in the production of both biogas and ethanol. It reduces the efficiency of the fermentation, increased fermenation time and increased cost biofuel.
  • Gram-positive bacteria particularly lactic acid bacteria (l_AB) are the most common bacterial contaminants found in ethanol production. See“The Alcohol Textbook” - a reference for the beverage, fuel and industrial alcohol industries”, 4 th edition, edited by KA Jaques, TP Lyons and DR Kelsall, 2003, Nottingham University Press, more particularly“Bacterial contamination and control in ethanol production” by N.V. Narendranath, pages 287-298.
  • a common solution to combat bacterial contamination is the use of virginiamycin, which is a streptogramin antibiotic similar to pristinamycin and quinupristin/dalfopristin.
  • virginiamycin which is a streptogramin antibiotic similar to pristinamycin and quinupristin/dalfopristin.
  • the ethanol industry is moving away from use of antibiotics as it increases cost. Furthermore, it may end up in the DDGS.
  • the extent of microbial contamination depends on the hygiene of the ethanol plant and the organic material. When plants are run in a hygienic fashion, antibiotics may not be necessary at all, or to a lesser extent, but because antibiotics are typically added at the start of the fermentation, it is likely that - on average - too much antibiotics are added.
  • antibiotics are typically added prophylactically, i.e. at the start of the fermentation. Therefore, there is a need to reduce the use of antibiotic in biofuel production.
  • the invention provides a process for producing a valuable compound through anaerobic fermentation of a composition comprising an organic material, said process comprising:
  • an antimicrobial compound is fed to the fermentation reactor when the concentration of lactic acid in the composition medium exceeds a pre-determined value.
  • a lactic acid sensitive sensor to control the feeding of antimicrobial compound enables real-time measurement of lactic acid, and on demand feeding of antimicrobial compound. This results in an overall reduction of antibiotics because lower amounts can be dosed. Furthermore, they are added only when necessary (i.e. in case of bacterial contamination) and prevents feeding antibiotics in cases where it would not be necessary.
  • the feeding of the antimicrobial compound can be done manually, or it can be controlled by a means for controlling the feed of the antimicrobial compound, which means can be a programmable means which is operatively connected to the lactic acid sensitive sensor.
  • An example of a suitable sensor is a biosensor, for example based on an enzyme such as lactate oxidase, which enzyme is preferably immobilized.
  • a particularly suitable sensor includes a voltammetric sensor.
  • the invention provides a process for producing a valuable compound through anaerobic fermentation of a composition comprising an organic material, said process comprising:
  • an antimicrobial compound is fed to the fermentation reactor when the concentration of lactic acid in the composition medium exceeds a pre-determined value.
  • the inventor has realized that measuring the lactic acid concentration in real time, by using a lactic acid sensitive sensor which is in direct contact with the fermentation medium, the dosing of antibiotics can be reduced to only those instances where there is bacterial growth.
  • This “on demand” addition of antimicrobial compound prevents feeding antibiotics in cases where it would not be necessary.
  • an antimicrobial compound is added prophylactically, i.e. at the start of the fermentation, lactic acid is always formed, and typically increases over time. A possible cause for this is that the antimicrobial compound is not stable at the fermentation temperature and degrades, thereby allowing bacteria to grow.
  • the antimicrobial compound can be dosed at a lower concentration because it is dosed when contamination levels are still low. In the early stages of contamination, when the bacteria have not yet grown out fully and when the lactic acid concentration is therefore low, a low dosage antimicrobial compound is sufficient to control the outgrow of the bacteria.
  • “a” and“an” are used herein to refer to one or to more than one (i.e. to one or at least one) of the grammatical object of the article.
  • “a fermentation reactor” may mean one fermentation reactor or more than one fermentation reactors.
  • the fermentation reactor may be an industrial type and scale fermentor. It may also be a shakeflask, bottle, or any other container, as long as it has an inlet and an outlet and at least one feed inlet for feeding an antimicrobial compound into the reactor, and can be used for anaerobic fermentation.
  • the inlet and outlet may be the same. For example, if the fermentation is carried out in a bottle, the opening of the bottle serves both as inlet and outlet.
  • the lactic acid sensitive sensor (henceforward referred to as“sensor”) is able to detect at least L-lactic acid.
  • the sensor When the sensor is in contact with the composition comprising an organic material (also referred to as“fermentation medium”) it sensor may give a reading e.g. in the form of a value which is displayed on a screen.
  • the sensitivity of the sensor is such that it is preferably able to detect concentrations of at least 0.01 g/L lactic acid.
  • the senor is able to distinguish between the presence or absence of lactic acid, whereby the sensor preferably has a threshold sensitivity which is below the pre-determined value.
  • Such sensor may give a“yes/no” reading (“no” meaning no lactic acid;“yes” meaning the presence of lactic acid).
  • the senor is able to provide the concentration of lactic acid. This concentration may be displayed on a screen or monitor.
  • anaerobic fermentation is defined as a fermentation which is run in the absence of oxygen or in which substantially no oxygen is consumed, preferably less than about 5, about 2.5 or about 1 mmol/L/h, more preferably 0 mmol/L/h is consumed (i.e. oxygen consumption is not detectable), and includes micro-anaerobic (or micro-aerobic) fermentation.
  • the feeding of the antimicrobial compound is done manually.
  • a person may monitor the reading of the sensor and, when the reading exceeds a predetermined value, add an antimicrobial compound via the feed inlet for feeding an antimicrobial compound into the reactor, for instance by using a scoop or by emptying a container.
  • the feeding of the antimicrobial compound can be controlled by a means for controlling the feed of the antimicrobial compound.
  • a means for controlling the feed of the antimicrobial compound can include a pump or screw which is able to feed antimicrobial compound via the feed inlet for feeding an antimicrobial compound into the reactor.
  • said means for controlling the feed of the antimicrobial compound is programmable and operatively connected to the sensor.
  • feeding of the antimicrobial compound is based on a reading of the sensor to which it is operatively connected.
  • the means is operated such that when the concentration of lactic acid exceeds a pre-determined value, it triggers a response which results in feeding of antimicrobial compound. This enables the use of in-line, on demand feeding of antimicrobial compound.
  • “operatively connected” means the sensor and the means for controlling the feed of the antimicrobial compound are connected in a way to perform the designated function, in casu feeding of antimicrobial compound when the concentration of lactic acid in the fermentation medium exceeds a pre-determined value.
  • the senor is a biosensor which is preferably based on an enzyme such as lactate oxidase, which enzyme is preferably immobilized.
  • the senor is a voltammetric sensor.
  • Voltammetric (or amperomeric) sensors are known in the art, see e.g. Kavita Rathee et at, Biosensors based on electrochemical lactate detection: A comprehensive review (2016), Biochemistry and Biophysics Reports, 5, 35-54.
  • a preferred electrochemical biosensor is based on the enzyme lactic acid oxidase (LOX or LOD) or lactic acid dehydrogenase (LDH or LD), most preferably lactic acid oxidase.
  • LOD lactic acid oxidase
  • LDH or LD lactic acid dehydrogenase
  • Rhathee et al extensively describe the principles of amperometric detection of lactic acid and how to immobilise the enzyme.
  • Voltammetric sensors have the advantage that they are able to provide the concentration of lactic acid.
  • Voltammetric sensors also have the advantage that the speed of formation of lactic acid can be monitored, i.e. the kinetics of lactic acid formation. Therefore, the invention is understood to include an embodiment wherein lactic acid is fed to the fermentation reactor when lactic acid is formed at a rate (g/L/h) which exceeds a pre-determined value.
  • the pre-determined value can be between 0.01 and 20 g/L lactic acid (relative to the volume of fermentation media). Ideally this value is as low as possible as this will give the most sensitive and immediate response to bacterial contamination.
  • the pre-determined value may be between 0.02 and 5 g/L, or between 0.03 and 1 g/L, or between 0.05 and 0.5 g/L, or at or around 0.1 g/L. The skilled person may select a higher value in the event that the composition comprising an organic material already comprises some lactic acid before the fermentation has started.
  • Suitable antimicrobial compounds include erythromycin, tylosin, virginiamycin, formaldehyde, nisin, penicillin, chlorine dioxide, peracetic acid, hops acids, and mixtures thereof. Lactrol is typically used at concentrations around 1 ppm, and a typical range is 0.5-2.0 ppm. However, sometimes much higher concentrations are used, up to 20 ppm when ethanol plants are struggling with infections.
  • the skilled person knows how much antimicrobial compound is to be fed when the concentration of lactic acid in the fermentation medium exceeds a pre-determined value.
  • Factors which determine the amount of feeding include the scale of the fermentor, the stage of the fermentation, the type of antimicrobial compound, and the recommended dose of the antimicrobial compound which is used.
  • the organic material comprises lignocellulosic biomass or hydrolysate thereof, such as a corn stover hydrolysate or a corn fiber hydrolysate.
  • hydrolysate a polysaccharide-comprising material (such as corn stover, corn starch, corn fiber, or lignocellulosic material, which polysaccharides have been depolymerized through the addition of water to form mono and oligosaccharide sugars. Hydrolysates may be produced by enzymatic or acid hydrolysis of the polysaccharide-containing material.
  • Lig nocellulose herein includes hemicellulose and hemicellulose parts of biomass.
  • Lig nocellulose includes lignocellulosic fractions of biomass. Suitable lignocellulosic materials may be found in the following list: orchard primings, chaparral, mill waste, urban wood waste, municipal waste, logging waste, forest thinnings, short-rotation woody crops, industrial waste, wheat straw, oat straw, rice straw, barley straw, rye straw, flax straw, soy hulls, rice hulls, rice straw, corn gluten feed, oat hulls, sugar cane, corn stover, corn stalks, corn cobs, corn husks, switch grass, miscanthus, sweet sorghum, canola stems, soybean stems, prairie grass, gamagrass, foxtail; sugar beet pulp, citrus fruit pulp, seed hulls, cellulosic animal wastes, lawn clippings, cotton, seaweed, trees, softwood, hardwood, poplar, pine,
  • Lignocellulose which may be considered as a potential renewable feedstock, generally comprises the polysaccharides cellulose (glucans) and hemicelluloses (xylans, heteroxylans and xyloglucans). In addition, some hemicellulose may be present as glucomannans, for example in wood-derived feedstocks.
  • glucans polysaccharides cellulose
  • hemicelluloses xylans, heteroxylans and xyloglucans
  • some hemicellulose may be present as glucomannans, for example in wood-derived feedstocks.
  • the enzymatic hydrolysis of these polysaccharides to soluble sugars, including both monomers and multimers, for example glucose, cellobiose, xylose, arabinose, galactose, fructose, mannose, rhamnose, ribose, galacturonic acid, glucuronic acid and other hexoses and pentoses occurs under the action of different enzymes
  • pectins and other pectic substances such as arabinans may make up considerably proportion of the dry mass of typically cell walls from non-woody plant tissues (about a quarter to half of dry mass may be pectins).
  • Lignocellulosic material may be pre-treated.
  • Pretreatment may comprise exposing the lignocellulosic material to an acid, a base, a solvent, heat, a peroxide, ozone, mechanical shredding, grinding, milling or rapid depressurization, or a combination of any two or more thereof.
  • This chemical pretreatment is often combined with heat- pretreatment, e.g. between 150-220°C for 1 to 30 minutes.
  • the organic material comprises starch or hydrolysate thereof, such as a corn starch hydrolysate.
  • Such pre-treated material is commonly subjected to enzymatic hydrolysis to release sugars that may be fermented according to the invention.
  • This may be executed with conventional methods, e.g. contacting with cellulases, for instance cellobiohydrolase(s), endoglucanase(s), beta-glucosidase(s) and optionally other enzymes.
  • the conversion with the cellulases may be executed at ambient temperatures or at higher temperatures, at a reaction time to release sufficient amounts of sugar(s).
  • the result of the enzymatic hydrolysis is hydrolysis product comprising C5/C6 sugars, herein designated as the sugar composition.
  • composition comprising organic material may also comprise sludge or biomass from purification, fermentation or digestion processes, or manure.
  • the fermentation in step (c) is preferably run at a temperature that is optimal for the yeast.
  • the fermentation process is performed at a temperature which is less than about 50°C, less than about 42°C, or less than about 38°C, preferably at a temperature which is lower than about 35°C, about 33°C, about 30 or about 28°C and at a temperature which is higher than about 20°C, about 22°C, or about 25°C.
  • the valuable compound may be ethanol or biogas, preferably ethanol.
  • the process optionally comprises recovering the valuable product.
  • existing technologies can be are used.
  • Existing methods of recovering ethanol from aqueous mixtures commonly use fractionation and adsorption techniques.
  • a beer still can be used to process a fermented product, which contains ethanol in an aqueous mixture, to produce an enriched ethanol- containing mixture that is then subjected to fractionation (e.g., fractional distillation or other like techniques).
  • fractionation e.g., fractional distillation or other like techniques
  • the fractions containing the highest concentrations of ethanol can be passed through an adsorber to remove most, if not all, of the remaining water from the ethanol.
  • the yeast may be recycled.
  • the yeast is capable to convert a C6 sugar and/or a C5 sugar.
  • C5 sugars also referred to as“pentose” or“pentose sugar”
  • pentose sugar are xylose, ribose, and arabinose, or derivatives thereof.
  • C6 sugars also referred to as“hexose” or“hexose sugar” are glucose, galactose, and mannose, preferably glucose and galactose, more preferably glucose.
  • the yeast is selected from the list consisting of Saccharomyces, Kluyveromyces, Candida, Scheffersomyces, Pichia, Schizosaccharomyces, Hansenula, Ogataea, Kloeckera, Schwanniomyces, Issatchenkia (such as I. orientalis) and Yarrowia, preferably the yeast is Saccharomyces cerevisiae.
  • a preferred yeast is a Saccharomyces, preferably S. cerevisiae.
  • the invention further provides a set up for running an anaerobic fermentation process suitable for producing a valuable compound comprising:
  • a fermentation reactor having an inlet and outlet and at least one feed inlet for feeding an antimicrobial compound into the reactor;
  • a lactic acid sensitive sensor which is capable of making contact with a fermentation medium when in progress
  • lactic acid sensitive sensor is operatively connected to the programmable means for feeding the antimicrobial compound into the reactor.
  • A“set up” may be an ethanol or biogas fermentation plant, or part thereof. It may consist of a single fermentation reactor, or two or more reactors, which may be operated in series or parallel.
  • the invention further provides the use of a lactic acid sensitive sensor to control the dosage of an antimicrobial compound in a set up for running an anaerobic fermentation process suitable for producing a valuable compound.
  • a lactic acid sensitive sensor to control the dosage of an antimicrobial compound in a set up for running an anaerobic fermentation process suitable for producing a valuable compound. All embodiments relating to the elements of the process of the invention such as the sensor, the fermentation reactor, the antimicrobial compound, the means for controlling the feed of said antimicrobial compound into the reactor, how the means and the sensor may be linked, and the valuable compound as described above in the process of the invention equally apply to the use of the invention.
  • a series of shake flasks is prepared with corn starch hydrolysate as fermentation medium.
  • To shake flasks B to G lactrol is added according to Table 1 .
  • Shake flasks 1 and 2 are connected to a three-electrode amperometric lactate biosensor as used in, and described by Imani et at,“A wearable chemical-electrophysiological hybrid biosensing system for real-time health and fitness monitoring”, Nature Communications (2016) DOI: 10.1038/ncomms1 1650.
  • the sensor is placed such that it is in contact with the fermentation medium and can be read by visual inspection.
  • the sensor is operatively connected to a programmable pump system which is able to dose lactrol to the shake flask.
  • the pump connected to shake flask 1 is programmed such that lactrol is dosed to the shake flask when the lactic acid concentration exceeds 0.1 g/L lactic acid.
  • the pump connected to shake flask 2 is programmed such that lactrol will be dosed to the shake flask when the lactic acid concentration exceeds 0.01 g/L lactic acid. If the lactic acid concentration exceeds 0.1 g/L (for shake flask 1 ) or 0.01 g/L (for shake flask 2) the pump starts feeding lactrol in an amount corresponding to 0.1 ppm. When this amount of lactrol has been fed, the pump shuts off until the lactic acid concentration again exceeds 0.1 g/L or 0.01 g/L.
  • the shake flasks are inoculated with Ethanol Red, a commercially available ethanol yeast and incubated at 32°C for 48 hours. After 48 hours of fermentation, the amount of lactic acid and ethanol, and the bacterial plate count are determined by methods known in the art. Results are in Table 1.
  • alpha-amylase An effective amount of alpha-amylase is added to 800 gram water and mixed well. Next, 200 gram corn flour is added to the aqueous mixture. The pH is adjusted the pH 5.5 with 2N H2SO4 or 4M KOH. The obtained mixture is transferred into a 2-litre bottle and capped. Thereafter, the bottle is incubated at 80°C in an oven or water bath for 4 hours under constant mixing. Then, the bottle is cooled to room temperature.
  • glucoamylase a commercially available ethanol yeast
  • Ethanol Red a commercially available ethanol yeast

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Abstract

L'invention concerne un procédé de production d'un composé de valeur par fermentation anaérobie d'une composition comprenant un matériau organique, ledit procédé comprenant (a) la fourniture d'un réacteur de fermentation ayant une entrée et une sortie, et au moins une entrée d'alimentation pour introduire un composé antimicrobien dans le réacteur, (b) la fourniture d'un capteur sensible à l'acide lactique qui est capable d'entrer en contact avec la composition ; (c) la fermentation, dans ledit réacteur de fermentation, de ladite composition en présence d'une levure qui est capable de convertir un sucre en C6 et/ou un sucre en C5 ; et éventuellement (d) la récupération du composé de valeur, un composé antimicrobien étant introduit dans le réacteur de fermentation lorsque la concentration en acide lactique dans le milieu de composition dépasse une valeur prédéterminée. L'utilisation d'un capteur sensible à l'acide lactique permet une mesure en temps réel de l'acide lactique, et une alimentation sur demande en composé antimicrobien. Il en résulte une réduction globale de l'utilisation d'antibiotiques car des quantités inférieures peuvent être dosées.
PCT/EP2019/055629 2018-03-09 2019-03-07 Procédé de fermentation anaérobie amélioré WO2019170788A1 (fr)

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EP18160855 2018-03-09

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WO2017087915A1 (fr) * 2015-11-20 2017-05-26 Duke University Biocapteurs de lactate et leurs utilisations
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