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WO2011069065A1 - Appareil et méthode de traitement de microorganismes pendant la propagation, le conditionnement et la fermentation utilisant du dioxyde de chlore et des extraits d'acides du houblon - Google Patents

Appareil et méthode de traitement de microorganismes pendant la propagation, le conditionnement et la fermentation utilisant du dioxyde de chlore et des extraits d'acides du houblon Download PDF

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WO2011069065A1
WO2011069065A1 PCT/US2010/058886 US2010058886W WO2011069065A1 WO 2011069065 A1 WO2011069065 A1 WO 2011069065A1 US 2010058886 W US2010058886 W US 2010058886W WO 2011069065 A1 WO2011069065 A1 WO 2011069065A1
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stream
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acid extract
hops acid
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Allen Ziegler
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Resonant Biosciences LLC
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Resonant Biosciences LLC
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    • 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
    • C12N1/005—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 after treatment of microbial biomass not covered by C12N1/02 - C12N1/08
    • 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
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • 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

Definitions

  • the present technology relates generally to anaerobic and aerobic microbial propagation, conditioning and/or fermentation.
  • the present technology involves a method of reducing the concentration of undesirable microorganisms while simultaneously encouraging propagation and/or conditioning of desirable microorganisms and increasing the efficiency of desirable microorganisms during fermentation.
  • Microorganisms such as yeast, fungi and bacteria, are used to produce a number of fermentation products, such as industrial grade ethanol, distilled spirits, beer, wine, pharmaceuticals and nutraceuticals (foodstuff that provides health benefits, such as fortified foods and dietary supplements).
  • fermentation products such as industrial grade ethanol, distilled spirits, beer, wine, pharmaceuticals and nutraceuticals (foodstuff that provides health benefits, such as fortified foods and dietary supplements).
  • Yeast are also commonly utilized in the baking industry.
  • Yeast are the most commonly used microorganism in fermentation processes. Yeast are minute, often unicellular, fungi. They usually reproduce by budding or fission.
  • Saccharomyces cerevisia the species predominantly used in baking and fermentation.
  • Non-Sacharomyces yeasts also known as non-conventional yeasts, are also used to make a number of commercial products.
  • Some examples of non- conventional yeasts include Kuyberomyces lactis, Yarrowia lipolytica, Hansenula polymorpha and Pichia pastoris.
  • cellulosic ethanol production production of ethanol from cellulosic biomass, utilizes fungi and bacteria.
  • these cellulolytic fungi include Trichoderma reesei and Trichoderma viride.
  • Trichoderma reesei Trichoderma viride.
  • a bacteria used in cellulosic ethanol production is Clostridium Ijungdahlii.
  • yeast used in distilleries and fuel ethanol plants are purchased from manufacturers of specialty yeasts.
  • the yeast are manufactured through a propagation process. Propagation involves growing a large quantity of yeast from a small lab culture of yeast. During propagation, the yeast are provided with the oxygen, nitrogen, sugars, proteins, lipids and ions that are necessary or desirable for optimal growth through aerobic respiration.
  • the yeast can undergo conditioning.
  • the objective of both propagation and conditioning is to deliver a large volume of yeast to the fermentation tank with high viability, high budding and a low level of infection by other microorganisms.
  • conditioning is unlike propagation in that it does not involve growing a large quantity from a small lab culture.
  • conditions are provided to re-hydrate the yeast, bring them out of hibernation and allow for maximum anaerobic growth and reproduction.
  • yeast Following propagation or conditioning, the yeast enter the fermentation process.
  • the yeast are combined in an aqueous solution with fermentable sugars.
  • the yeast consume the sugars, converting them into aliphatic alcohols, such as ethanol.
  • yeast can become contaminated with bacteria or other undesirable microorganisms. This can occur in one of the many vessels used in propagation, conditioning or fermentation. This includes propagation tanks, conditioning tanks, starter tanks, fermentations tanks, piping and heat exchangers between these units.
  • Bacterial or microbial contamination reduces the fermentation product yield in three main ways.
  • the sugars that could be available for yeast to produce alcohol are consumed by the bacteria or other undesirable microorganisms and diverted from alcohol production.
  • the end products of bacterial metabolism such as lactic acid and acetic acid, inhibit yeast growth and yeast fermentation/respiration, which results in less efficient yeast production.
  • the bacteria or other undesirable microorganisms compete with the yeast for nutrients other than sugar.
  • bacteria or other undesirable microorganisms can grow much more rapidly than the desired yeast.
  • the bacteria or other microorganisms compete with the yeast for fermentable sugars and retard the desired bio-chemical reaction resulting in a lower product yield.
  • Bacteria also produce unwanted chemical by-products, which can cause spoilage of entire fermentation batches. Removing these bacteria or other undesirable microorganisms allows the yeast to thrive, which results in higher efficiency.
  • Some previous methods of reducing bacteria or other undesirable microorganisms during propagation, conditioning and fermentation take advantage of the higher temperature and pH tolerance of yeast over other microorganisms. This is done by applying heat to or lowering the pH of the yeast solution. However, these processes are not entirely effective in retarding bacterial growth. Furthermore, the desirable yeast microorganisms, while surviving, are stressed and not as vigorous or healthy. Thus, the yeasts do not perform as well.
  • Another approach involves washing the yeast with phosphoric acid. This method does not effectively kill bacteria and other microorganisms. It can also stress the yeast, thereby lowering their efficiency.
  • antibiotics have previously been added to yeast propagation, conditioning or fermentation batch to neutralize bacteria. Fermentation industries typically apply antibiotics to conditioning, propagation and fermentation processes. Antibiotic dosage rates range between 0.1 to 3.0 mg/L and generally do not exceed 6 mg/L.
  • Antibiotic residues and establishment of antibiotic-resistant strains is a global issue. These concerns may lead to future regulatory action against the use of antibiotics.
  • One area of concern is distillers grain that is used for animal feed. European countries do not allow the byproducts of an ethanol plant to be sold as animal feed if antibiotics are used in the facility. Distiller grain sales account for up to 20% of an ethanol plant earnings. Antibiotic concentration in the byproduct can range from 1-3% by weight, thus negating this important source of income.
  • Fermentation plants can experience infections. This occurs when undesirable microorganism levels increase to above a normal or allowable level. This can occur due to process design, poor quality feed stock or other contributing factors. When this occurs antibiotic usage is usually increased to compensate for the infection. These conditions instigate overuse of antibiotic which can stress yeast and impact efficiency or cause regulatory non-compliance.
  • Chlorine dioxide (C10 2 ) has many industrial and municipal uses. When produced and handled properly, C10 2 is an effective and powerful biocide, disinfectant and oxidizer.
  • C10 2 has been used as a disinfectant in the food and beverage industries, wastewater treatment, industrial water treatment, cleaning and disinfections of medical wastes, textile bleaching, odor control for the rendering industry, circuit board cleansing in the electronics industry, and uses in the oil and gas industry. It is an effective biocide at low concentrations and over a wide pH range. C10 2 is desirable because when it reacts with an organism in water, it reduces to chlorite ion and then to chloride, which studies to date have shown does not pose a significant adverse risk to human health.
  • brewers added an aqueous 2-6% by weight sodium chlorite solution, otherwise known as stabilized chlorine dioxide, to their fermentation batches in an attempt to kill bacteria and other microorganisms.
  • sodium chlorite reacts in an acidic environment it can form C10 2 .
  • the C10 2 added using this method was not substantially pure, which made it difficult to ascertain the amount added or control that amount with precision. If the amount is not precisely maintained, the C10 2 can kill the desired yeast or inhibit the glucoamylase enzyme that is present to prepare the fermentable sugars. If these undesirable consequences occur, the addition of C10 2 will not result in more efficient production. This method is also not effective at a neutral or basic pH level.
  • C10 2 gas for treating yeast during the propagation, conditioning and/or fermentation process is desirable because there is greater assurance of C10 2 purity when in the gas phase.
  • C10 2 is, however, unstable in the gas phase and will readily undergo decomposition into chlorine gas (Cl 2 ), oxygen gas (0 2 ), and heat.
  • the high reactivity of C10 2 generally requires that it be produced and used at the same location.
  • Chlorine dioxide is a bactericide with a much greater degree of efficacy than hops acid. Hops acids utilized in the recommended dosage rates act as a bacteriastatic agent, effective against gram positive bacteria only - not a bactericide as chlorine dioxide does.
  • An embodiment of the current method for reducing undesirable microorganism concentration, promoting yeast propagation, and increasing yeast efficiency in an aqueous fluid stream comprises (a) introducing a quantity of fermentable carbohydrate to an aqueous fluid stream, (b) introducing a quantity of yeast to the aqueous fluid stream, (c) generating C10 2 gas, (d) dissolving the C10 2 gas to form a C10 2 solution, (e) introducing an aqueous C10 2 solution into the aqueous fluid stream, and (f) introducing a hops acid extract stream into the aqueous fluid stream. These steps can be performed sequentially or in a different order.
  • the "undesirable" microorganisms intended to be reduced are those that compete for nutrients with the desirable microorganisms, such as yeast and Trichoderma that promote in the fermentation processes involved here.
  • the aqueous C10 2 and hops acid extract solution employed in the present method does not appear to detrimentally affect the growth and viability of desirable, fermentation-promoting microorganisms, but does appear to eliminate or at least suppress the growth of undesirable microorganisms that interfere with the fermentation process.
  • the elimination or suppression of undesirable microorganisms appears to have a favorable effect on the growth and viability of desirable microorganisms, for the reasons set forth in the Background section.
  • the C10 2 gas can be generated by reacting chlorine gas with water and then adding sodium chlorite.
  • the C10 2 gas could be generated by reacting sodium hypochlorite with an acid and then adding sodium chlorite.
  • the C10 2 gas can also be generated by reacting sodium chlorite and hydrochloric acid.
  • the C10 2 gas can also be generated using electrochemical cells and sodium chlorate or sodium chlorite. Equipment- based generation could also be used to create C10 2 gas using sodium chlorate and hydrogen peroxide.
  • the C10 2 solution has a concentration of less than about 15 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm. In another embodiment the C10 2 solution has a concentration of between about 5 and about 50 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm. In one embodiment the C10 2 solution has an efficiency as C10 2 in the stream of at least about 90%. As used in this application "to have an efficiency as C10 2 of at least about 90%" means that at least about 90% of the C10 2 solution or C10 2 gas is in the form of C10 2 .
  • Another embodiment of the current method that reduces undesirable microorganism concentration, promotes yeast propagation, and increases yeast efficiency in an aqueous fluid stream comprises (a) introducing a quantity of fermentable carbohydrate to an aqueous fluid stream, (b) introducing a quantity of yeast to the aqueous fluid stream, (c) introducing C10 2 having an efficiency as C10 2 of at least about 90% into the aqueous fluid stream and (d) introducing a hops acid extract into the aqueous fluid stream. These steps can be performed sequentially or in a different order.
  • the C10 2 having an efficiency as C10 2 in the stream of at least about 90% can be produced by equipment or non-equipment based methods.
  • non-equipment based methods of C10 2 generation include dry mix chlorine dioxide packets that include both a chlorite precursor packet and an acid activator packet.
  • Equipment-based methods include using electrochemical cells with sodium chlorate or sodium chlorite, and a sodium chlorate/hydrogen peroxide method.
  • the C10 2 solution is in the form of an aqueous solution having a concentration of less than about 15 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm.
  • the C10 2 solution is in the form of an aqueous solution having a concentration of between about 5 and about 50 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm.
  • the C10 2 is in a gaseous form.
  • An embodiment of the current apparatus for reducing undesirable microorganisms, promoting producing microorganism propagation, and increasing efficiency comprises a C10 2 generator, a batch tank, a hops acid extract tank and a process vessel for containing an aqueous microorganism solution.
  • the C10 2 generator comprises an inlet for introducing at least one chlorine-containing feed chemical and an outlet for exhausting a C10 2 gas stream from the generator.
  • the batch tank is fluidly connected to the C10 2 generator outlet and receives the C10 2 gas stream from the C10 2 generator outlet.
  • the batch tank comprises an inlet for introducing a second water stream and an outlet for exhausting an aqueous C10 2 solution from the batch tank.
  • the process vessel is fluidly connected to the batch tank.
  • the process vessel is also fluidly connected to the hops acid extract tank. In operation, introducing the C10 2 and hops acid extract to the process vessel promotes propagation of producing microorganisms present in the vessel.
  • the batch tank preferably has an inlet for introducing a second water stream and an outlet for exhausting an aqueous C10 2 solution.
  • the batch tank is capable of exhausting an aqueous C10 2 solution that has a concentration of less than about 5,000 mg/L.
  • the exhausted C10 2 solution is dosed to have a concentration between about 5 and about 50 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm.
  • the exhausted C10 2 solution is dosed to have a concentration of less than about 15 mg/L and the hops acid extract has a concentration of from about 0.1 to about 5 ppm.
  • the process vessel can be a conditioning tank, heatable, capable of performing liquefaction or a yeast propagation vessel.
  • the process vessel could also be a fermentation tank having an inlet for producing microorganisms, an inlet for water, an inlet for fermentation chemicals and an outlet for the fermentation product connecting to processing equipment.
  • Another embodiment of the current method for reducing undesirable microorganism concentration, promoting desirable microorganism propagation, and increasing desirable microorganism efficiency in an aqueous fluid stream comprises (a) introducing a quantity of cellulose to an aqueous fluid stream, (b) introducing a quantity of desirable microorganisms to the aqueous fluid stream, (c) generating C10 2 gas, (d) dissolving the C10 2 gas to form a C10 2 solution, (e) introducing an aqueous C10 2 solution into the aqueous fluid stream and (f) introducing an aqueous hops acid extract stream into the aqueous fluid stream.
  • These steps can be performed sequentially or in a different order.
  • the C10 2 solution has an efficiency as C10 2 in the stream of at least about 90%.
  • Another embodiment of the current method that reduces undesirable microorganism concentration, promotes desirable microorganism propagation, and increases desirable microorganism efficiency in an aqueous fluid stream comprises (a) introducing a quantity of cellulose to an aqueous fluid stream, (b) introducing a quantity of desirable microorganisms to the aqueous fluid stream, (c) introducing C10 2 having an efficiency as C10 2 of at least about 90% into the aqueous fluid stream and (d) introducing an aqueous hops acid extract stream into the aqueous fluid stream.
  • the concentrations of bacteria and other undesirable microorganisms are reduced while simultaneously propagation and/or conditioning of desirable microorganisms is encouraged, and the efficiency of those desirable microorganisms in fermentation and an apparatus for carrying out this method increased.
  • chlorine dioxide was determined to be effective at reducing the concentration of bacteria and other undesirable microorganisms while simultaneously encouraging propagation and/or conditioning of desirable microorganisms and increasing the efficiency of those desirable microorganisms in fermentation.
  • Isomerized alpha extract is used as an example throughout this application.
  • hops acid extracts could be used.
  • beta acid compounds, alpha acids, isomerized alpha acids, rho isomerized alpha acids, tetra isomerized alpha acids, hexa isomerized alpha acids and hop leaf could be used.
  • Plant scale evaluations have determined that adding a small amount of hops acid extract, for example about .1 to about 5 ppm in addition to and simultaneously with chlorine dioxide results in a synergistic effect.
  • the addition of chlorine dioxide and hops acid extract simultaneously results in improved microbiology efficacy, enhanced ethanol production, reduced glycerol formation and increased yeast viability and propagation.
  • yeast fermentation The production of fuel ethanol by yeast fermentation is used as an example. However, this is merely one illustration and should not be understood as a limitation.
  • Other fermentation products could include distilled spirits, beer, wine, pharmaceuticals, pharmaceutical intermediates, baking products, nutraceuticals (foodstuff that provides health benefits, such as fortified foods and dietary supplements), nutraceutical intermediates and enzymes.
  • the current method could also be utilized to treat yeast used in the baking industry.
  • Other fermenting microorganisms could also be substituted such as the fungi and bacteria typically used in cellulosic ethanol production, Trichoderma reesei, Trichoderma viride, and Clostridium Ijungdahlii.
  • the fermentation process begins with the preparation of a fermentable carbohydrate.
  • corn is one possible fermentable carbohydrate.
  • Other carbohydrates including cereal grains and cellulose-starch bearing materials, such as wheat or milo, could also be substituted.
  • Cellulosic biomass such as straw and cornstalks could also be used.
  • Cellulosic ethanol production has recently received attention because it uses readily available nonfood biomass to form a valuable fuel.
  • corn-based ethanol production the corn is ground into a fine powder called meal.
  • the meal is then mixed with water and enzymes, such as alpha-amylase, and passed through a cooker in order to liquefy the starch.
  • a secondary enzyme such as glucoamylase
  • glucoamylase will also be added to the mash to convert the liquefied starch into a fermentable sugar.
  • the glucoamylase cleaves single molecules of glucose from the short chain starches, or dextrins. The glucose molecules can then be converted into ethanol during fermentation.
  • Yeast small microorganisms capable of fermentation, will also be added to the corn mash.
  • Yeast are fungi that reproduce by budding or fission.
  • Saccharomyces cerevisia the species predominantly used in baking and fermentation.
  • Non-Sacharomyces yeasts also known as non-conventional yeasts, are naturally occurring yeasts that exhibit properties that differ from conventional yeasts.
  • Non- conventional yeasts are utilized to make a number of commercial products such as amino acids, chemicals, enzymes, food ingredients, proteins, organic acids, nutraceuticals, pharmaceuticals, cosmetics, polyols, sweeteners and vitamins.
  • non- conventional yeasts include Kuyberomyces lactis, Yarrowia lipolytica, Hansenula polymorpha and Pichia pastoris.
  • the current methods and apparatus are applicable to intermediates and products of both Sacharomyces and non-conventional yeast.
  • yeast slurry Most of the yeast used in fuel ethanol plants and other fermentation processes are purchased from manufacturers of specialty yeast.
  • the yeast are manufactured through a propagation process and usually come in one of three forms: yeast slurry, compressed yeast or active dry yeast.
  • Propagation involves growing a large quantity of yeast from a small lab culture of yeast. During propagation the yeast are provided with the oxygen, nitrogen, sugars, proteins, lipids and ions that are necessary or desirable for optimal growth through aerobic respiration.
  • the yeast may undergo conditioning.
  • the objectives of both propagation and conditioning are to deliver a large volume of yeast to the fermentation tank with high viability, high budding and a low level of infection by other microorganisms.
  • conditioning is unlike propagation in that it does not involve growing a large quantity from a small lab culture.
  • conditions are provided to re-hydrate the yeast, bring them out of hibernation and allow for maximum anaerobic growth and reproduction.
  • yeast Following propagation or conditioning, the yeast enter the fermentation process.
  • the glucoamylase enzyme and yeast are often added into the fermentation tank through separate lines as the mash is filling the fermentation tank. This process is known as simultaneous saccharification and fermentation or SSF.
  • SSF simultaneous saccharification and fermentation
  • the yeast produce energy by converting the sugars, such as glucose molecules, in the corn mash into carbon dioxide and ethanol.
  • the fermentation mash, now called “beer” is distilled.
  • This process removes the 190 proof ethanol, a type of alcohol, from the solids, which are known as whole stillage. These solids are then centrifuged to get wet distillers grains and thin stillage.
  • the distillers grains can be dried and are highly valued livestock feed ingredients known as dried distillers grains (DDGS).
  • the thin stillage can be evaporated to leave a syrup.
  • the alcohol is passed through a dehydration system to remove remaining water. At this point the product is 200 proof ethanol.
  • This ethanol is then denatured by adding a small amount of denaturant, such as gasoline, to make it unfit for human consumption.
  • the propagation, conditioning and fermentation processes can be carried out using batch and continuous methods.
  • the batch process is used for small-scale production. Each batch is completed before a new one begins.
  • the continuous fermentation method is used for large-scale production because it produces a continuous supply without restarting every time.
  • the current method and apparatus are effective for both methods.
  • the mash or the fermentation mixture can become contaminated with other microorganisms, such as spoilage bacteria, wild yeast or killer yeast.
  • microorganisms compete with the yeast for fermentable sugars and retard the desired bio-chemical reaction resulting in a lower product yield. They can also produce unwanted chemical by-products, which can cause spoilage of entire fermentation batches. Wild yeast are a primary concern in the beverage industry because they can cause taste and odor problems with the final product. Killer yeast produce a toxin that is lethal to the desired alcohol producing yeast.
  • Producers of ethanol attempt to increase the amount of ethanol produced from one bushel of cereal grains, which weigh approximately 56 pounds (25.4 kilograms). Contamination by microorganisms lowers the efficiency of yeast making it difficult to attain or exceed the desired levels of 2.8-2.9 gallons per bushel (.42-.44 liters per kilogram). Reducing the concentration of microorganisms will encourage yeast propagation and/or conditioning and increase yeast efficiency making it possible to attain and exceed these desired levels.
  • C10 2 solution has many uses in disinfection, bleaching and chemical oxidation. Yeast can withstand and indeed thrive in a C10 2 environment. However, bacteria, wild yeasts, killer yeasts and molds will succumb to the properties of CIO 2 allowing the producing, desirable yeast to thrive and achieve higher production.
  • CIO 2 can be added at various points in the propagation, conditioning and/or fermentation processes to kill unwanted microorganisms and promote growth and survival of the desirable microorganisms.
  • This CIO 2 can be added as an aqueous solution or a gas.
  • the C10 2 can be added during propagation, conditioning and/or fermentation.
  • the CIO 2 solution can be added to cook vessels, fermentation tanks, propagation tanks, conditioning tanks, starter tanks or during liquefaction.
  • the CIO 2 solution can also be added to the interstage heat exchange system or heat exchangers.
  • the C10 2 has an efficiency as C10 2 in the stream of at least about 90%. Adding C10 2 having a known purity allows for addition of a controlled amount of C10 2 .
  • hops acid extract are useful for killing bacteria, wild yeasts, killer yeasts and molds while allowing yeast or other producing microorganisms to survive and thrive. Fermentation industries typically apply hops acid extracts to propagation and fermentation. Typically, hops acid extract dosage rates range between 15 and 50 ppm as active product when utilized independently.
  • Plant scale evaluations have determined that adding a small amount of hops acid extract, for example about .1 to about 5 ppm, in addition to and simultaneously with chlorine dioxide results in a synergistic effect.
  • the addition of chlorine dioxide and hops acid extract simultaneously results in improved microbiology efficacy, enhanced ethanol production, reduced glycerol formation and increased yeast viability and propagation.
  • Evaluations were conducted at a fuel ethanol facilities utilizing chlorine dioxide and the hops acid extract.
  • the facility dosed chlorine dioxide to the propagator, fermentor and interstage heat exchanger at a dosage rate of between about 1 and about 50 mg/L.
  • Hops acid extract (Iso-Alpha extract 30%) was simultaneously applied to the propagator and fermentor at a rate of between about 0.1 and about 5 ppm.
  • Ethanol efficiency in the plant was increased by addition of chlorine dioxide and hops acid extract simultaneously. An unexpected result was that no detrimental or inhibitory effect was noted between chlorine dioxide and hops acid extract.
  • a fermentation ethanol plant was treated for approximately six months with chlorine dioxide and hops acid. Chlorine dioxide was added at numerous application points. Chlorine dioxide was applied at the interstage heat exchangers at 15 ppm/2.51gpm/5.03 lb/hr. Chlorine dioxide was applied to the fermentation process during phase one at 14 ppm/2.53 gpm/5.06 lb/hr at 16 hours. Chlorine dioxide was added to the fermentation process during phase two at 3 ppm/.36 gpm/.72 lb/hr. Chlorine dioxide was added to the yeast propagator at 15 ppm/.13 gpm/.25 lb/hr at 6.5 hours.
  • Chlorine dioxide was added to the cook water at 3 ppm/.27 gpm/.4 lb/hr. Chlorine dioxide was added to the CIP at 10 ppm/.27 gpm/.4 lb/hr. This makes the total chlorine dioxide addition 13.45 lb/hr.
  • Chlorine dioxide was found to be synergistic with isoalpha hops acid.
  • a 1.5 ppm dosage of isoalpha hops acid in conjunction with 13 lb/hr of chlorine dioxide produced similar efficacy with regard to ethanol production as would typically be achieved with over 30-50 ppm of isalpha hops acid utilized alone or 20 lb/hour chlorine dioxide used alone.
  • the hops acid extract can be added simultaneously with the chlorine dioxide at the various points in the propagation, conditioning and/or fermentation processes where chlorine dioxide was previously added.
  • the hops acid extract can be added to cook vessels, fermentation tanks, propagation tanks, conditioning tanks, starter tanks or during liquefaction.
  • the hops acid extract solution can also be added to the interstage heat exchange system or heat exchangers.
  • C10 2 and hops acid extract can be added directly into the fermentation mixture. This can be done by adding the C10 2 and hops acid extract in conjunction with the yeast and glucoamylase, for example during the SSF stage. Chlorine dioxide dosages of less than about 15 mg/L, preferably less than about 10 mg/L and most preferably less than about 7.5 mg/L used with hops acid extract dosages of between .5 and 5 ppm can be added directly into the fermentation mixture.
  • the C10 2 and hops acid extract can also be added to the mash prior to the fermentation process, for example before the SSF stage.
  • Chlorine dioxide dosages of between about 10 and about 75 mg/L, preferably between about 10 and about 50 mg/L and most preferable between about 20 and about 50 mg/L used with hops acid extract dosages of between .5 and5 ppm can be added to the mash prior to fermentation.
  • Chlorine dioxide and hops acid extract can also be added during propagation and/or conditioning.
  • C10 2 can be added to the yeast slurry before SSF replacing the acid washing step.
  • Chlorine dioxide dosages of less than about 50 mg/L used with hops acid extract dosages of between .5 and 6 ppm can be added during propagation and/or conditionaing.
  • C10 2 gas can decompose explosively, it is typically produced on-site.
  • C10 2 gas can be produced using electrochemical cells and a sodium chlorite or sodium chlorate solution.
  • An equipment based sodium chlorate/hydrogen peroxide method also exists.
  • non-equipment based binary, multiple precursor dry or liquid precursor technologies can be used. Examples of non-equipment based methods of C10 2 generation include dry mix chlorine dioxide packets that include both a chlorite precursor packet and an acid activator packet.
  • hypochlorous acid reacts with water to form hypochlorous acid and hydrochloric acid. These acids then react with sodium chlorite to form chlorine dioxide, water and sodium chloride.
  • sodium hypochlorite is combined with hydrochloric or other acid to form hypochlorous acid. Sodium chlorite is then added to this reaction mixture to produce chlorine dioxide.
  • the third method combines sodium chlorite and sufficient hydrochloric acid.
  • the C10 2 gas produced is between 0.0005 and 5.0 % by weight in air.
  • the C10 2 gas is dissolved in a solvent in order to create a C10 2 solution.
  • C10 2 gas is readily soluble in water.
  • the water and C10 2 gas are combined in quantities that create a solution for application directly to the fermentation mixture, with a concentration of less than about 15 mg/L, preferably less than about 10 mg/L, and most preferably less than about 7.5 mg/L.
  • the water and C10 2 gas are combined in quantities that create a solution for application to the corn mash prior to fermentation, with a concentration of between about 10 and about 75 mg/L, preferably between about 10 and about 50 mg/L, and most preferable between about 20 and about 50 mg/L.
  • the water and C10 2 gas are combined in quantities that create a solution for application to the yeast during propagation with a concentration of less than about 50 mg/L.
  • the C10 2 solution has an efficiency as C10 2 in the stream of at least about 90%.
  • Pure or substantially pure C10 2 is desirable because it allows the user to precisely maintain the amount of C10 2 added to the yeast. (The single term “pure” will be used hereinafter to mean either pure or substantially pure.) If too little C10 2 is added the dosage will not be effective in killing undesirable microorganisms. If too much C10 2 is added it can kill the desired yeast. If either of these situations occurs, the addition of C10 2 will not result in more efficient ethanol production. Addition of pure C10 2 allows the user to carefully monitor and adjust the amount of C10 2 added to the yeast. This enables the user to add adequate C10 2 to assure microbial efficacy without killing the yeast.
  • C10 2 is also desirable for another reason.
  • Glucoamylase enzyme is important in ethanol production to convert short chain starches (or dextrins) into fermentable glucose molecules.
  • C10 2 does not exhibit a significant reaction with glucoamylase.
  • C10 2 can reduce to form chlorite ion.
  • the chlorite ion can inhibit the glucoamylase enzyme at approximately 14 mg/L and above. Inhibition of glucoamylase enzyme can lower ethanol production.
  • a chlorite ion concentration of 14 mg/L can be produced by a C10 2 dosage rate of about 50 to 60 mg/L. Addition of pure C10 2 allows the user to add dosage rates below the level where glucoamylase inhibition can occur.
  • the C10 2 and hops acid extract solution is introduced at some point during the production of ethanol.
  • the C10 2 and hops acid extract solution can be added during propagation, conditioning and/or fermentation.
  • the C10 2 and hops acid extract solution can also be added directly to the corn mash.
  • the C10 2 and hops acid extract solution can be added to cook vessels, fermentation tanks, propagation tanks, conditioning tanks, starter tanks or during liquefaction.
  • the C10 2 and hops acid extract solution can also be added to the piping between these units or heat exchangers.
  • C10 2 and hops acid extract can also be used simultaneously to achieve improved results in the production of cellulosic ethanol.
  • Cellulosic ethanol is a type of ethanol that is produced from cellulose, as opposed to the sugars and starches used in producing carbohydrate based ethanol.
  • Cellulose is present in non-traditional biomass sources such as switch grass, corn stover and forestry. This type of ethanol production is particularly attractive because of the large availability of cellulose sources.
  • Cellulosic ethanol by the very nature of the raw material, introduces higher levels of contaminants and competing microorganism into the fermentation process. C10 2 and hops acid extract used simultaneously could be particularly helpful in cellulosic ethanol production as an antimicrobial agent.
  • the cellulose in the chemical hydrolysis method can be treated with dilute acid at high temperature and pressure or concentrated acid at lower temperature and atmospheric pressure. In the chemical hydrolysis process the cellulose reacts with the acid and water to form individual sugar molecules. These sugar molecules are then neutralized and yeast fermentation is used to produce ethanol. C10 2 and hops acid extract could be used during the yeast fermentation portion of this method as outlined above.
  • Enzymatic hydrolysis can be carried out using two methods.
  • the first is known as direct microbial conversion (DMC).
  • DMC direct microbial conversion
  • This method uses a single microorganism to convert the cellulosic biomass to ethanol.
  • the ethanol and required enzymes are produced by the same microorganism.
  • CIO 2 and hops acid extract could be used during the propagation/conditioning or fermentation steps with this specialized organism.
  • the second method is known as the enzymatic hydrolysis method.
  • cellulose chains are broken down using cellulase enzymes. These enzymes are typically present in the stomachs of ruminants, such as cows and sheep, to break down the cellulose that they eat.
  • the cellulose is made via fermentation by cellulolytic fungi such as Trichoderma reesei and Trichoderma viride.
  • the enzymatic method is typically carried out in four or five stages.
  • the cellulose is pretreated to make the raw material, such as wood or straw, more amenable to hydrolysis.
  • the cellulase enzymes are used to break the cellulose molecules into fermentable sugars.
  • the sugars are separated from residual materials and added to the yeast.
  • the hydrolyzate sugars are fermented to ethanol using yeast.
  • the ethanol is recovered by distillation.
  • the hydrolysis and fermentation can be carried out together by using special bacteria or fungi that accomplish both processes. When both steps are carried out together the process is called sequential hydrolysis and fermentation (SHF).
  • SHF sequential hydrolysis and fermentation
  • C10 2 and hops acid extract can be introduced for microbiological efficacy at various points in the enzymatic method of hydrolysis.
  • C10 2 and hops acid extract could be used in the production, manufacture and fermentation of cellulase enzymes made by Trichoderma and other fungi strains.
  • the C10 2 and hops acid extract can be added in the cellulosic simultaneous saccharification and fermentation phase (SSF).
  • SSF cellulosic simultaneous saccharification and fermentation phase
  • the C10 2 and hops acid extract can be introduced in the sequential hydrolysis and fermentation (SHF) phase. They could also be introduced at a point before, during or after the fermentation by cellulolytic fungi that create the cellulase enzymes.
  • the CIO 2 and hops acid extract could be added during the yeast fermentation phase, as discussed above.
  • the gasification process does not break the cellulose chain into sugar molecules.
  • the carbon in the cellulose is converted to carbon monoxide, carbon dioxide and hydrogen in a partial combustion reaction.
  • the carbon monoxide, carbon dioxide and hydrogen are fed into a special fermenter that uses a microorganism such as Clostridium Ijungdahlii that is capable of consuming the carbon monoxide, carbon dioxide and hydrogen to produce ethanol and water.
  • the ethanol is separated from the water in a distillation step.
  • CIO 2 and hops acid extract could be used as an antimicrobial agent in the fermentation step involving microorganisms such as Clostridium Ijungdahlii that are capable of consuming carbon monoxide, carbon dioxide and hydrogen to produce ethanol and water.
  • the apparatus has a C10 2 generator.
  • the C10 2 generator has an input for electricity. There is also an inlet for at least one chlorine containing chemical.
  • the C10 2 generator should also have an outlet for exhausting a C10 2 gas stream from the generator. In one embodiment the C10 2 gas stream exiting the generator is between 0.0005 and 5.0 % by weight in air.
  • a batch tank that receives the C10 2 gas stream is fluidly connected to the C10 2 generator outlet.
  • the C10 2 gas is dissolved in water to form a C10 2 solution.
  • the batch tank has an inlet for introducing a water stream.
  • the water stream and the C10 2 gas stream are combined to form a C10 2 solution.
  • the concentration of the C10 2 solution in the batch tank can vary across a wide range. Concentrations of up to about 5,000 mg/L can be achieved and concentrations of up to about 8,000 mg/L can be achieved with additional equipment.
  • the C10 2 solution is then exhausted from the batch tank through an outlet at a specified dosage rate to create a solution of the desired concentration.
  • the dosed C10 2 solution for application directly to the fermentation mixture, has a concentration of less than about 15 mg/L, preferably less than about 10 mg/L, and most preferable less than about 7.5 mg/L.
  • the dosed C10 2 solution, for application to the corn mash prior to fermentation has a concentration of between about 10 and about 75 mg/L, preferably between about 10 and about 50 mg/L, and most preferable between about 20 and about 50 mg/L.
  • the dosed C10 2 solution, for use in propagation has a concentration of less than about 50 mg/L.
  • the exiting C10 2 solution has an efficiency as C10 2 in the stream of at least about 90%.
  • the apparatus has a hops acid extract tank.
  • hops acid extract such as isomerized alpha extract
  • the concentration of the hops acid extract solution in the batch tank can vary across a wide range.
  • the hops acid extract solution is then exhausted from the batch tank through an outlet at a specified dosage rate to create a solution of the desired concentration.
  • the dosed hops acid extract solution has a concentration between about .1 and 5 ppm.
  • the hops acid extract tank is typically a pre -mix tank.
  • a process vessel containing an aqueous microorganism solution is fluidly connected to the batch tank and the hops acid extract tank via outlets on the batch tank and hops acid extract tank.
  • the process vessel could be a cook vessel, fermentation tank, conditioning tank, starter tank, propagation tank, liquefaction vessel and/or piping or heat exchanger between these units. Introducing the C10 2 and hops acid extract solution into the process vessel is capable of promoting propagation of producing microorganism present while simultaneously decreasing the concentration of undesirable microorganisms.
  • skid-mounted equipment For smaller scale production of fermentation products, skid-mounted equipment is ideal. Skid mounting allows the equipment to be manufactured off site, shipped to the desired location and easily installed. This ensures ease in transportation, faster erection and commissioning.
  • the C10 2 generator, batch tank, hops acid extract tank, process vessel and connecting equipment could be made in a skid-mounted fashion.

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Abstract

L'invention porte sur un méthode permettant de réduire la concentration en microorganismes indésirables, de favoriser la propagation /et le conditionnement de microorganismes désirables, et d'accroître l'efficacité de microorganismes désirables dans des flux de liquide aqueux, cette méthode consistant: a) à introduire une quantité d'hydrate de carbone de sucre ou de cellulose fermentescible dans un flux de liquide aqueux; (b) à introduire une quantité de microorganismes désirables dans le flux de liquide aqueux; (c) à introduire une solution de ClO2 dans le flux de liquide aqueux; et (f) à introduire un extrait d'acides du houblon dans ledit flux de liquide aqueux. L'invention porte également sur un appareil à cet effet comportant: un générateur de ClO2, une cuve de fermentation, un réservoir d'extrait d'acides du houblon, et une cuve de traitement. L'introduction dans la cuve de traitement du ClO2 et de l'extrait d'acides du houblon, provenant respectivement de la cuve de fermentation et du réservoir d'extrait d'acides, favorise la propagation des microorganismes obtenus présents dans la cuve.
PCT/US2010/058886 2009-12-04 2010-12-03 Appareil et méthode de traitement de microorganismes pendant la propagation, le conditionnement et la fermentation utilisant du dioxyde de chlore et des extraits d'acides du houblon Ceased WO2011069065A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
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WO2006015071A1 (fr) * 2004-07-29 2006-02-09 Pureline Treatment Systems, Llc Generateur de solution de dioxyde de chlore
WO2007097874A1 (fr) * 2006-02-22 2007-08-30 Resonant Biosciences, Llc Appareil et procédé pour traiter des micro-organismes pendant leur propagation, leur conditionnement, et leur fermentation

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WO2006015071A1 (fr) * 2004-07-29 2006-02-09 Pureline Treatment Systems, Llc Generateur de solution de dioxyde de chlore
WO2007097874A1 (fr) * 2006-02-22 2007-08-30 Resonant Biosciences, Llc Appareil et procédé pour traiter des micro-organismes pendant leur propagation, leur conditionnement, et leur fermentation

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RAMIREZ-OROZCO MARTIN ET AL: "DEBARYOMYCES HANSENII GROWTH IN NONSTERILE SEAWATER CLO2-PEPTONE-CONTAINING MEDIUM DEBARYOMYCES HANSENII GROWTH IN NONSTERILE SEAWATER CLO2-PEPTONE-CONTAINING MEDIUM", CANADIAN JOURNAL OF MICROBIOLOGY, NRC RESEARCH PRESS, CA, vol. 47, no. 7, 1 July 2001 (2001-07-01), pages 676 - 679, XP008078385, ISSN: 0008-4166, DOI: DOI:10.1139/CJM-47-7-676 *
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12330968B2 (en) 2020-12-29 2025-06-17 Ecolab Usa Inc. Method for controlling odor and taste producing metabolites in water systems through use of primary and secondary oxidation processes

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