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WO2004108609A1 - Milieu de fermentation comprenant des eaux usees et son utilisation - Google Patents

Milieu de fermentation comprenant des eaux usees et son utilisation Download PDF

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Publication number
WO2004108609A1
WO2004108609A1 PCT/DK2004/000385 DK2004000385W WO2004108609A1 WO 2004108609 A1 WO2004108609 A1 WO 2004108609A1 DK 2004000385 W DK2004000385 W DK 2004000385W WO 2004108609 A1 WO2004108609 A1 WO 2004108609A1
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WIPO (PCT)
Prior art keywords
fermentation
wastewater
process according
biomass
waste
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
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PCT/DK2004/000385
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English (en)
Inventor
Anne Belinda Thomsen
Helene Bendstrup Klinke
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Riso National Laboratory
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Riso National Laboratory
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Filing date
Publication date
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Priority to EP20040736046 priority Critical patent/EP1641713A1/fr
Publication of WO2004108609A1 publication Critical patent/WO2004108609A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/28—Anaerobic digestion processes
    • 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
    • Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel

Definitions

  • the present invention relates in general to the field of producing a fermentation product.
  • a novel process for the production of a fermentation product such as lactic acid, combustible fuel product e.g. ethanol or cell biomass by using wastewater derived from e.g. municipal sewage, slaughterhouse waste, household waste or manure as a source of water and nutrient supply for a hydrolysis process and/or a fer- mentation process.
  • the carbon dioxide content in some industrial districts is as high as ten times normal. Acid forming pollutants have been found in some instances to increase the acidity of rainwater from its normal pH of about 6.9 to values of 4.0. Rainwater having a pH of 5.5 or lower will destroy aquatic life and can do substantial harm to buildings, monuments, and other structures.
  • One proposal is to subject the sewage to dry distillation or pyrolysis at temperatures between about 900°C and about 1000°C without the injection of steam or oxygen, thereby attempting to produce crude oil. Scrubbing the gas products of this process is required and the scrubbing treatment causes pollution of water and the atmosphere.
  • Another proposal is to include the step of applying heat externally to a retort in which sewage is heated without internal combustion.
  • the sewage is distilled or pyrolysed at a tern- perature of about 400°C.
  • the resulting gas is cracked, enriched, and scrubbed to make it suitable for heating and ligating purposes.
  • scrubbing of the gas creates pollution problems and about 30% of the carbon remain unused and will be discarded with the ash.
  • Another proposal calls for sewage to be destructively distilled while on a travelling grate in an open system.
  • Organic material is thermally decomposed at temperatures between about 450°C and about 1100°C in the absence of oxygen. Some 34-36% of the starting material remains at the completion of the process. Most of the gas produced during the process is consumed in the process and the gas which is not so consumed is heavily di- luted with carbon dioxide and nitrogen and is not suitable for use in the chemical industry or release to the atmosphere.
  • stack heights have been increased to affect better dispersion of pollutants.
  • Increasing stack heights add to the cost of constructing and maintaining stacks, yet provides no solution to the underlying problem, i.e., emission of harmful substances such as sulphur oxides, chlorine gases, phosphor oxides, etc.
  • a specific aspect of sewage disposal relates to separation and handling of the aqueous phase of sewage in the following referred to as wastewater. Not much attention has been given to the disposal problems associated with wastewater.
  • Wastewater derived from sewage comprises a range of compounds such as dissolved organic matter (proteins, sugars, fats etc), nutrient salts, minerals, and metals, thus, with- out an efficient cleaning technology, it becomes a hazard for the environmental stability.
  • COD Chemical Oxygen Demand
  • Other types of compounds present in wastewater are compounds such as sodium, potassium, magnesium, calcium, sulphur, phosphorus, nitrogen, iron, copper, iodine, fluorine, chlorine cobalt etc.
  • Nutrients like nitrogen and phosphorous serve as nutrients for a range of microorganisms. These compounds have been linked to dangerous toxic microorganisms such as Pfisteria piscicida. Pfisteria is believed to be responsible for major fish kills and disease events in several Mid-Atlantic States and may pose a risk to human health. Nitrogen and/or phosphorus stimulate aquatic algae growth, thus depleting water bodies of oxygen and killing fish and other aquatic organisms. Nutrient pollution comes from runoff of excess fertilisers, industrial wastewater, municipal wastewater, animal sewage, and other diffuse sources, as well as from wastewater treatment plants and some industries.
  • US 6,267,309 disclose a method for producing ethanol or other chemicals from the organic portion of a waste stream of municipal solid sewage.
  • the content of heavy metals present in the organic portion is reduced by treating the solid sewage with sulphuric acid or via ionic exchange.
  • the treated organic portion is subsequently shredded followed by acidification and thermal treatment.
  • the resulting composition can be utilised in a fermentation process.
  • DE 19946299 discloses a method for fermenting readily decomposable solid sewage products and difficult-to-decompose solid sewage products to produce methane, which method includes the following steps: 1) the readily decomposable sewage products and the diffi- cult-to-decompose sewage products are mixed with water in two separate vessels and heated for 1 hour at 70 °C; 2) the difficult-to-decompose sewage products are mixed with an enzyme suspension and subjected to aerobic hydrolysis for 3 days and subsequently mixed with the readily decomposable sewage products; 3) the mixture then undergoes an anaerobic hydrolysis and acidogenesis for 3 days followed by separation of the formed sediment from the remaining liquid of the anaerobic hydrolysis and acidogenesis; 4a) the formed sediment are mixed with enzyme, 10% paper sewage and 5% whey and subjected to aerobic hydrolysis, and 4b) the remaining liquid is mixed with sewage and used as a medium for the production of methane.
  • methanogens are capable of producing fermentable products more or less independent on the medium if just any kind of energy source and nutrients are present. This characteristics makes the methanogens useful in cleaning procedures of sewage, furthermore they are naturally occurring in sewage and grow well under the anaerobic conditions present in sewage.
  • specific fermentation products such as ethanol, is provided by substrate specific organisms which require a more specific and/or well defined constituents of energy source and nutrients, therefore wastewater has not until now been suggested as a source for the fermentation of specific fermentation products.
  • the disadvantage of the methods in the prior art is that the wastewater is discarded after a suitable cleaning procedure and as such, makes no use of the nutrients, minerals and the water present in the collected wastewater in a fermentation, but mainly the solid phase of the sewage is utilised. Furthermore, additional handling steps are required in or- der to provide a suitable solid phase, which is usable as carbon source and in which the amount of heavy metals is removed or substantially reduced.
  • the present invention discloses a suitable process and medium utilising wastewater derived from sewage for the fermentation of useful products, such as ethanol. Additionally, it has been found possible to replace nutrients, minerals and/or water conventionally added to fermentation media as pure compounds by using wastewater derived from sewage. Thereby, it has become possible to provide a cheap and efficient process and medium for the production of fermentation products.
  • a process for production of fermentation products comprises the steps of:
  • step (ii) adding a biomass material to the wastewater of step (i) to obtain a biomass slurry
  • step (iii) subjecting the slurry of step (ii) to a fermentation process to obtain a fermentation product
  • step (iv) separating the fermentation product resulting from step (iii).
  • a fermentation medium comprising wastewater is provided.
  • a still further aspect relates to the use of a fermentation medium according to the invention for the production of a fermentation product.
  • the process of the present invention involves the use of waste- water derived from sewage, which, prior to use may optionally be subjected to any kind of degradation or digestion to release nutrients, minerals, organic material into the waste- water.
  • a digestion comprises the steps of thermal treatment and/or a optionally fermentation by aerobic or anaerobic digestion.
  • the wastewater is separated from the solid phase.
  • the wastewater is then mixed with a carbohydrate source to provide a biomass slurry which is mixed with a microorganism capable of producing the product of interest.
  • the product produced by the fermentation is then separated and optionally isolated and purified.
  • the present invention also discloses a fermentation medium comprising wastewater, which thereby limit the additional supplementation of nutrients, minerals and/or water.
  • This fermentation medium is useful in the production of a fermentation product in the petrochemical industry, pharmaceutical industry, biotech industry, chemical industry, and food and feed industry.
  • the sewage may be separated into two phases (i) an aqueous phase (wastewater) and (ii) a solid phase.
  • the separation of the two phases is as described later.
  • wastewater refers to any type of discarded aqueous phase derived from sewage.
  • the wastewater may comprise a community's used water and water carried solids, including used water from industrial processes such as the pharmaceutical, food and feed industry, that flow to a sewage treatment plant. Whey, storm wa- ter, surface water and groundwater infiltration that enters a wastewater treatment plant may also be included in the term "wastewater”.
  • sewage relates to the waste, i.e. liquid and solid phase, 5 discarded such as municipal waste, household waste, slaughterhouse waste, e.g. meat/bone flour and blood, garbage and/or industrial waste such as waste from the food, feed and pharmaceutical industry, and waste derived from animal farming, such as manure and solid manure, and from gardening.
  • waste i.e. liquid and solid phase
  • 5 discarded such as municipal waste, household waste, slaughterhouse waste, e.g. meat/bone flour and blood, garbage and/or industrial waste such as waste from the food, feed and pharmaceutical industry, and waste derived from animal farming, such as manure and solid manure, and from gardening.
  • waste i.e. liquid and solid phase
  • 5 discarded such as municipal waste, household waste, slaughterhouse waste, e.g. meat/bone flour and blood, garbage and/or industrial waste such as waste from the food, feed and pharmaceutical industry, and waste derived from animal farming, such as manure and solid manure, and from gardening.
  • the sewage may be collected from
  • the wastewater is derived from sewage by separating the aqueous phase (the wastewater) from the solid phase.
  • the supply of nutrients, minerals and/or water 15 constitutes a significant part of the costs involved in the fermentation process.
  • the present inventors surprisingly found that naturally occurring water, nutrients and minerals originally present in the wastewater are preserved and used in a fermentation process, in the present context also referred to as "the mandatory fermentation process". In this way the process and the fermentation medium according to the present 20 invention becomes much cheaper than conventionally used processes and fermentation media and relieve the pressure on the environment.
  • the sewage and/or the wastewater may be subjected to a treatment, as discussed below, for the release of nutrients and/or minerals as well as for
  • the wastewater comprises an increased amount of nutrients and/or minerals compared to untreated wastewater, such as at least 5% more nutrients and/or minerals, e.g such as at least 10% such as at least
  • the wastewater may be used with or without any further treatment for removing nutrients and minerals.
  • the wastewater is obtained directly from the sewage discharge source or it can be obtained after the sewage has been subjected to at least one optionally treatment to remove solids, microorganisms, chemicals and/or enzymes. Treatments for removing nutrients and in ⁇ hibitory substances are discussed later.
  • the content of ammonium in wastewater is in the range of 500-10,000 mg/l, such as 1,000-8,000 mg/l, e.g. 1,500-6,000 mg/l, such as 2,000-2,500 mg/l, such as 2,000-4,000 mg/l.
  • the content of phosphor in wastewater is in the range of 25-5,000 mg/l, such as 50-3,000 mg/l, e.g. 80-2,000 mg/l, such as 90- 1,000 mg/l, such as 150-200 mg/l, e.g. 100-500 mg/l.
  • the content of chemical oxygen demand (COD) in wastewater is in the range of 100-250,000 mg/l, such as 150-150,000 mg/l, e.g. 200-100,000 mg/l, such as 300-75,000 mg/l, e.g. 350-50,000 mg/l, such as 400-25,000 mg/l, e.g. 500-10,000 mg/l, such as 1,000-10,000 mg/l, e.g. 10,000-100,000 mg/l, such as 15,000-90,000 mg/l, e.g. 25,000-80,000 mg/l, such as 50,000-75,000 mg/l.
  • COD chemical oxygen demand
  • the content of ammonium, phosphor and chemical oxygen demand (COD), in wastewater resulting from thermal treatment, fermentation by anaerobe digestion and filtration is in the range of 2,000-4,000 mg/l, 100-500 mg/l and 1,000-10,000 mg/l, respectively.
  • the amount of any of ammonium, phosphor and chemical oxygen demand (COD), respectively resulting from thermal treatment is in the range of 2,000-4,000 mg/l, 100-500 mg/l and 50,000-75,000 mg/l, respectively.
  • the wastewater When separated from the solid phase, the wastewater is used, either alone or in combination with other solutions or substrates in microbial fermentation production of any kind of desired product.
  • the wastewater comprises at least one nutrient selected from the group consisting of nitrogen, phosphor, magnesium, zinc, manganese, cobalt, copper, calcium, iron, molybdenum, boron, urea, proteins, amino acids, a compound containing any of such elements and any combination hereof.
  • solutions or substrates relates to different types of me- dia or aqueous solutions with or without nutrients, minerals, carbohydrate-containing materials, vitamins, detergents, amino acids, lipids and salts.
  • the wastewater may be sterilised prior to being subjected to the mandatory fermentation process.
  • the wastewater is supplemented with at least one additional nutrient.
  • the nutrient used to supplement the wastewater is selected from the group consisting of urea, nitrogen, phosphor, magnesium, zinc, manganese, cobalt, copper, calcium, iron, molybdenum, boron, any compound containing any of such elements and any combination hereof.
  • the amount of the supplementary nutrient, if added, calculated on the final medium is typically at the most 10%, e.g. at the most 25%, such as at the most 50%, e.g. at the most 75%, such as at the most 90%, e.g. at the most 95%.
  • the remaining solid phase of sewage may be at least partially removed, e.g. by a process selected from the group consisting of filtration, centrifugation, sedimentation and decanting.
  • the sewage can be treated thermally, by ultra wave, enzymatically, by wet oxidation, steam explosion or combinations hereof. Such treatments are discussed below.
  • solids, microorganisms, chemicals or enzymes may be removed or deacti- vated from the wastewater by any conventional process for such removal or deactivation, such as sterilisation by e.g. heating, boiling or cooking, simple filtration, chromatography, microfiltration, diafiltration, centrifugation or neutralisation.
  • the heavy metals present in the waste- water need not be removed or reduced before the wastewater are used in the fermentation of a product of interest. Thus, no additional treatment is needed in order to remove heavy metals from the wastewater.
  • the biomass material is a carbohydrate-containing material (hexoses and pentoses) including a glucan and pentosan containing material such as a lignocellulosic material, starch containing material, cellulose, starch, an organic waste material, household wastes, paper materials, paper pulp, return paper, straw, maize stems, forestry waste (log slash, bark, small branches, twigs and the like), sawdust, wood-chips, simple monomeric sugars and molasse from sugar beet or sugar cane.
  • a carbohydrate-containing material hexoses and pentoses
  • a glucan and pentosan containing material such as a lignocellulosic material, starch containing material, cellulose, starch, an organic waste material, household wastes, paper materials, paper pulp, return paper, straw, maize stems, forestry waste (log slash, bark, small branches, twigs and the like), sawdust, wood-chips, simple
  • the carbohydrate-containing material used in the mandatory fermentation process of the invention is not present in the waste- water, but an additional carbohydrate-containing material is added to the fermentation mixture.
  • Carbohydrate containing materials that can be used in the mandatory process of the present invention include materials of directly fermentable sugars (e.g. molasses) starch containing materials as well as lignocellulosic materials of plant origin, the lignocellulose, which is the principal component of such materials, in general being built up predominantly of cellulose, hemicellulose and lignin.
  • directly fermentable sugars e.g. molasses
  • lignocellulose which is the principal component of such materials, in general being built up predominantly of cellulose, hemicellulose and lignin.
  • Cellulose which is a ⁇ -glucan comprising of anhydro D-glucose units, is the main structural component of plant cell walls and normally constitutes about 35-60% by weight (% w/w) of lignocellulosic materials.
  • Hemicellulose is the term used to denote non-cellulosic polysaccharides associated with cellulose in plant tissues. Hemicellulose typically constitutes about 20-35% w/w of lignocellulosic materials, and the majority of hemicelluloses consists predominantly of polymers based on pentose (five-carbon) sugar units, such as D-xylose and D-arabinose units, although minor proportions of hexose (six-carbon) sugar units, such as D-glucose and D- mannose units, are generally also present.
  • pentose (five-carbon) sugar units such as D-xylose and D-arabinose units
  • hexose (six-carbon) sugar units such as D-glucose and D- mannose units
  • Lignin a complex, cross-linked polymer based on variously substituted p-hydroxyphenyl- propane units, generally constitutes about 10-30% w/w of lignocellulosic materials. It is believed that lignin functions as a physical barrier to the direct bioconversion (e.g. by fer- menting microorganisms) of cellulose and hemicellulose in lignocellulosic materials which have not been subjected to some kind of pre-treatment process (which may suitably be a wet-oxidative process as described in the following) to disrupt the structure of lignocellulose.
  • pre-treatment process which may suitably be a wet-oxidative process as described in the following
  • biomass in the form of low-cost by-products from gardening such as garden refuse, waste materials from agriculture, forestry, the timber industry and the like.
  • processes of the invention are applicable to any kind of hemicellulose-con- taining lignocellulosic materials.
  • Relevant materials thus include wooden or non-wooden plant material in the form of stem, stalk, shrub, foliage, bark, root, sugar beet pulp, shell, pod, nut, husk, fibre, vine, straw, hay, grass, bamboo, sugar cane bagasse, or reed, singularly or in a mixture.
  • the ratio between the solid carbohydrate-containing material and the wastewater is in the range of 1:99-1: 1, preferably in the range of 1:49-1: 2, e.g. in the range of 1:9-1:4.
  • the initial proportion of carbohydrate- containing material in wastewater will be in the range of 0.02-1 kg/litre of wastewater, often 0.05-0.35 kg/litre, such as 0.05-0.25 kg/litre, depending on the form, bulk and/or dimensions of the lignocellulosic material.
  • the process of the invention at the highest practicable ratio between carbohydrate-containing material and wastewater i.e. at the highest ratio which permits adequate mixing of the carbohydrate-containing material in the wastewater comprising the oxidising agent and which leads to a satisfactorily high rate of degradation of carbohydrate-containing material.
  • additional water is added to the biomass slurry obtained in step (ii) of the present process.
  • the source of additional water can be tap water, distilled water and wastewater.
  • a grinding step e.g. milling, abrading, grinding, crushing, chopping, chipping or the like
  • enhancing e.g., the physical mobility, mixability, ratio of surface area to mass and the like of the material.
  • the slurry obtained in step (ii) contains, calculated on the total carbohydrate content, at the most 90% microbially femnent- able sugars, e.g. at the most 80% microbially fermentable sugars, such as at the most 70% microbially fermentable sugars, e.g. at the most 60% microbially fermentable sugars, such as at the most 50% microbially fermentable sugars, e.g. at the most 40% microbially fermentable sugars, such as at the most 25% microbially fermentable sugars and e.g. at the most 5% microbially fermentable sugars.
  • a suitable microorganism is selected depending on the product of interest.
  • the microorganism is selected from the group consisting of bacteria, yeast and fungi.
  • the mandatory fermentation process may be either an anaerobic fermentation process or an aerobic fermentation process or a combination hereof.
  • the mandatory fermentation process is substantially not a methane producing fermentation process.
  • the term "substantially” relates to that the mandatory fermentation at the most produces 1% methane, such as at the most 0.5%, including at the most 0.1%.
  • a suitable microorganism for this purpose includes a mesophilic microorganism (i.e. one which grows optimally at a temperature in the range of 20-40°C), e.g. a yeast including Saccharomyces cerevisiae, also referred to as "baker's yeast”.
  • any microorganism capable of converting xylose to ethanol can be used in the process according to the invention.
  • Useful microorganisms include e.g. certain types of thermophiles (i.e. organisms which grow optimally at an elevated temperature, typically a temperature in excess of about 50°C) and genetically engineered microorganisms derived therefrom.
  • a suitable organism for the ethanol fermentation is selected from the group consisting of Ther- moanaerobacter species including T. mathranii, Zymomonas species including Z. mobilis and yeast species such as Pichia species.
  • An example of a useful strain of T. mathranii is described in Sonne-Hansen et al. (1993) or Ahring et al. (1996) where said strain is designated strain A3M4.
  • a useful ethanol-fermenting organism can be selected from a genetically modified organism of one of the above useful organisms having, relative to the organism from which it is derived, an increased or improved ethanol-fermenting activity.
  • genetically modified bacterium is used in the conventional meaning of that term i.e. it refers to strains obtained by subjecting an organism to any conventionally used mutagenization treatment including treatment with a chemical muta- gen such as ethanemethane sulphonate (EMS) or N-methyl-N'-nitro-N-nitroguanidine (NTG), UV light or to spontaneously occurring mutants, including classical mutagenesis.
  • mutants of the above mentioned organism can be provided by such technology including site-directed mutagenesis and PCR techniques and other in vitro or in vivo modifications of specific DNA sequences once such sequences have been identified and isolated.
  • lactic acid or other desired products such as vitamins, antibiotics, amino acids and colours
  • the industrially most useful lactic acid bacteria are found among Lactococcus species, Streptococcus species, Lactobacillus species, Leuconostoc species, Pediococcus species and Brevibacte um species.
  • the strict anaerobes belonging to the genus Bifidobacterium is generally included in the group of lactic acid bacteria.
  • a group of lactic acid bacterial species which are used as so-called probiotics or used in the fermentation of probiotics include e.g.
  • Lactobacillus johnsonii Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus casei, Lactocoocus lactis subsp. cremoris, Lactobacillus paracasei subsp.
  • Lactobacillus rhamnosus Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus acidophilus (Lactocidin), Bifidobacterium infantis, Bifidobacterium adolescent ' s, Bifidobacterium longum, Bifidobacterium animalis, Bifidobacterium breve, Enterococcus faecium, Streptococcus saliva us and Streptococcus lactis (Nisin).
  • acetic acid species from Aetobactor spp. are useful or Closteridium thermoaceticum.
  • Closteridium thermoaceticum For the fermentation of propionate and butyrate Closte d- ium propionicum and Closteridium tyrobutyricum, respectively, can be used.
  • Caldicellulosiruptor saccharolyticus may be used in the production of hydrogen and different kind of species of Penicillium are useful in the production of various kinds of antibiotics.
  • vitamins organisms such as Propi- onibacteria shermanii (B12) and Corynebacterum sp. (vitamin C) are useful.
  • An example of an amino acid producing species is Corynebacterium glutamicum which produces the amino acid L-lysine and colours may be produced by Curvularia lunata (Anthraquinone).
  • filamentous fungi may be used such as Aspergillus niger, Botrytis cinerea, Penicillium brasilianum, Schizophyllum commune or Trichoderna reesei (Thygesen et al. 2003).
  • the production of the fermentation product is a continuous process.
  • the wastewater obtained from the production of a fermentation product according to the present invention may be recycled or at least part of the wastewater may be recycled.
  • This recycling is provided in order to reuse the wastewater as an aqueous liquid phase in the process of the invention thereby reducing the consumption of water and minimising the quantity of waste material emerging from the process.
  • This recycling may be processed mostly without any substantial inhibition of the pre-treatment of the lignocellulosic biomass or of the subsequently hydrolysis or the fermentation of sugars.
  • the mandatory fermentation process is performed using a mixed culture of organisms.
  • This mixed culture may comprise at least one type of microorganism capable of producing the product of interest and at least one type of microorganism capable of reducing the level of inhibitory substances. Examples of the latter type of organisms are disclosed below.
  • the mandatory process and the medium according to the present invention can be used for the fermentation production of any kind of product.
  • the fermentation product produced is selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins and enzymes.
  • the fermentation process and the fermentation medium according to the invention also can be useful in the production of cell biomass of a selected microorganism such as a bacteria, a fungi such as Mucor indicus or yeast, such as S. cere- visiae.
  • a selected microorganism such as a bacteria, a fungi such as Mucor indicus or yeast, such as S. cere- visiae.
  • the sewage, biomass or waste- water itself may by subjected to a treatment for the release of nutrients, minerals and/or water as well as for releasing any carbohydrate-containing material already present in the sewage, biomass or wastewater and making the carbohydrate-containing material more accessible for the microorganisms used in the mandatory fermentation.
  • a treatment of the sewage, biomass and wastewater can be a thermal treatment, an optionally fermentation/digestion, enzymatic treatment, acid hydrolysis, wet oxidation, steam explosion, filtration or any combination thereof.
  • the sewage or the wastewater obtained from sewage is subjected to any one of the following steps in any given order: (a) thermal treatment, (b) fermentation (anaerobic diges- tion), or (c) filtration.
  • a wet oxidation or elevated temperature treatment e.g. thermal treatment or steam explosion of carbohydrate-containing material may be used.
  • Such treatments are water consuming so it is an embodiment of the present invention to use the provided wastewater during such treatments.
  • wastewater during thermal treatment the wastewater can function directly as process water, due to this high temperature treatment the wastewater will also be sterilised.
  • steam explosion as described below water is mainly needed before a high temperature steam pre-treatment for presoaking and after a treatment as a fermentation medium; in this case, as well as other similar cases, the provided wastewater need to be sterilised before added to the treated biomass (as described later).
  • the biomass material is pretreated by thermal treatment, wet oxidation, steam explosion, dilute sulphuric acidic or other relevant acids, alkaline solutions, organic solvents or any combination hereof before being subjected to fermentation.
  • the pretreated, untreated biomass and/or the biomass slurry is hydrolysed by enzymatic treatment and/or chemical hydroly- sis, this chemical hydrolysis may either be by acidic treatment or by alkaline treatment.
  • wet oxidation and “wet-oxidative” as used herein refers to a process which takes place in an aqueous medium, i.e. sewage, wastewater, liquid water or a liquid medium containing at least a substantial proportion of water, in the presence of an oxidising agent which reacts oxidatively with one or more components or species present (as a solid or solids, and/or in dissolved form) in the medium.
  • an oxidising agent which reacts oxidatively with one or more components or species present (as a solid or solids, and/or in dissolved form) in the medium.
  • the process normally takes place at an elevated temperature, i.e.
  • Oxidising agent as mentioned above relates mainly, but is not limited, to oxygen or hydrogen peroxide that under suitable concentrations and under suitable conditions of temperature and reaction time are appropriate for use in a wet-oxidative process in the manner of the invention.
  • Hydrogen peroxide is highly soluble in water, is readily available commercially as aqueous solutions of concentration ranging from relatively dilute (e.g. hydrogen peroxide concentrations of around 3% or 5% w/w) to relatively concentrated (e.g. hydrogen peroxide concentrations of about 30-35% or 30-49 w/w) and is, like oxygen, a very acceptable oxidising agent from an environmental point of view.
  • the initial concentration of hydrogen per- oxide in the liquid, aqueous medium is typically in the range of 0.5-10% w/w.
  • oxygen When oxygen is used as oxidising agent, it is preferred that the process is performed in the presence of oxygen introduced at an initial partial pressure of oxygen equal to or exceeding the ambient partial pressure of oxygen (i.e. the partial pressure of oxygen in the surrounding air, which at sea level is normally around 0.2 bar, typically about 0.21 bar), and initial oxygen partial pressures which lie in the range from about 0.2 to about 35 bar will normally be of interest. It is, however, generally preferable to employ initial oxygen partial pressures of at least 0.5 bar, normally in the range of 0.5-35 bar. Typical initial partial pressures of oxygen will be in the range of 1-15 bar, such as 3-12 bar, e.g. 5-12 bar.
  • the solubility of oxygen in water at temperatures of relevance for the process of the invention increases with oxygen partial pressure, and the use of such elevated partial pressures of oxygen can thus be advantageous in ensuring the availability of sufficient oxygen in dissolved form.
  • the oxygen used may be added in the form of substantially pure oxygen or in the form of an oxygen-containing gas mixture (such as atmospheric air) which in addition to oxygen is constituted by one or more other gases (e.g. nitrogen and/or an inert gas, such as argon) that are not detrimental to the performance of the process of the invention; it will, how- ever, often be advantageous to use substantially pure oxygen (such as oxygen of >99% purity, which is commercially available in conventional gas cylinders under pressure).
  • an oxygen-containing gas mixture such as atmospheric air
  • other gases e.g. nitrogen and/or an inert gas, such as argon
  • Preferred conditions for wet oxidation and other thermal processes including dilute acid treatment include the use of temperatures in the vicinity of, or in excess of, •100°C.
  • temperatures in the range of 120-300°C, e.g. 120-240, such as 180-210°C, e.g. 180-240°C, more typically in the range of 180-220°C will be appropriate for the vast majority of such embodiments of the process according to the invention, and when using lignocellulosic materials of preferred types it will be usual to employ temperatures in the range of 160-210°C, such as 180-210°C. Good results appear to be obtainable with temperatures around 185-195°C or 170-190°C.
  • the temperature applied should be a temperature at which boiling of the liquid, aqueous medium does not occur under the pressure conditions in question.
  • the temperature is less than 220°C, such as less than 200°C, e.g. less than 195°C including less than 190°C, e.g. less than 185°C, such as less than 180°C including less than 175°C, such as less than 125°C, e.g. less than 100°C including less than 80°C, e.g. less than 70°C.
  • the temperature is 70°C, 80°C, 90°C, 100°, 110°C or 120°C.
  • the wet oxidation and the steam explosion convert a large portion of the biomass material to C0 2 , H 2 0 and simpler, more oxidised organic compounds, mainly low-molecular weight carboxylic acids.
  • a treatment or digestion in the present context also referred to as the “optionally fermentation/digestion” is performed using methane-producing microorganisms (also known as methanogens) which constitute a unique group of prokaryotes which are capable of forming methane from certain classes of organic substrates, methyl substrates (methanol, methylamine, dimethylamine, trimethylamine, methylmercaptan and dimethylsulfide) or acetate (sometimes termed acetoclastic substrate) under anaerobic conditions.
  • methane-producing microorganisms also known as methanogens
  • methyl substrates methanol, methylamine, dimethylamine, trimethylamine, methylmercaptan and dimethylsulfide
  • acetate sometimes termed acetoclastic substrate
  • inhibitory substances relates to substances produced during thermal treatment.
  • Such substances include carboxylic acids such as acetic acid, formic acid and lactic acid, and furans including 5-hydroxymethylfurfural, 2-furfural and 2- furoic acid and phenols including guaiacol, syringoi, 4-hydroxy benzalde-hyde, vanillin, sy- ringaldehyde, 3,4,5-trimethoxybenzaldehyde, 4-hydroxy aceto-phenone, acetovanillone, acetosyringone, 3,4,5-trimethoxyacetophenone, 4-hydroxy benzoic acid, vanillic acid, sy- ringic acid, p-coumaric acid and ferulic acid.
  • carboxylic acids such as acetic acid, formic acid and lactic acid
  • furans including 5-hydroxymethylfurfural, 2-furfural and 2- furoic acid and phenols including guaiacol, syringoi, 4-hydroxy benzal
  • Methanogens are found within various genera of bacteria, and methanogenic bacteria of relevance in the context of the present invention include species of Methanobacterium, Methanobrevibacter, Methanothermus, Methanococcus, Methanomicrobium, Methano- genium, Methanospirillum, Methanoplanus, Methanosphaera, Methanosarcina, Metha- nolobus, Methanoculleus, Methanothrix, Methanosaeta, Methanopyrus or Methanocor- pusculum; some of these, notably species of Methanopyrus, are highly thermophilic and can grow at temperatures in excess of 100°C. Only three genera of methanogenic bacteria, viz.
  • Methanosarcina, Methanosaeta and Methanothrix appear to contain species capable of carrying out the acetoclastic reaction, i.e. conversion of acetate to methane (and carbon dioxide).
  • useful methanogenic bacteria can be selected from a genetically modified bacterium of one of the above useful organism having, relative to the organism from which it is derived, an increased or improved methane producing activity. Such a genetically modified organism can be obtained by the methods discussed below.
  • Such other types of microorganisms include certain fermentative anaerobic bacteria capable of converting, for example, glucose to products such as acetate, propionate, butyrate, hydrogen and C0 2 , and so-called acetogenic bacteria, which convert organic substances such as propionate, butyrate and ethanol to acetate, formate, hydrogen and C0 2 .
  • the level of the above inhibitory substances, including nitrogen, phosphate or toxins, in the fermentation medium and/or wastewater is reduced by employing various kinds of plants in a plant clarifying unit capable of degrading or uptaking such substances.
  • a plant clarifying unit capable of degrading or uptaking such substances Any kinds of known plants capable of de- grading such inhibitory substances can be used, including water hyacinth (Eichornia crassipes) and St. Augustine Gras (Stenotaphrum secundatum).
  • the derived wastewater from the above treatments may be sterilised alone or together with the biomass to be pretreated and fermented.
  • paper pulp used as biomass material can be added without further modification to the sterilised wastewater together with enzymes and yeast.
  • the sterilisation can be performed by a wet oxidation process, steam explosion or dilute acid treatment resulting in subsequently at least partial hydrolysis of the cellulose and hemicellulose to obtain a slurry of wastewater and biomass containing an amount of microbially fermentable sugars as well as nutrients that permits the slurry or wastewater to be used as an ethanol fermentation medium.
  • hydrolysis treatment is to hydrolyse oligosaccharide and polysac- charide species as well as nutrient salts and substances during the pretreatment (e.g. steam explosion, wet oxidation and acidic hydrolysis) in step (ii) of cellulose and/or hemicellulose origin to form fermentable sugars (e.g. glucose, xylose and possibly other mono- saccharides).
  • pretreatment e.g. steam explosion, wet oxidation and acidic hydrolysis
  • fermentable sugars e.g. glucose, xylose and possibly other mono- saccharides.
  • Such treatments may be physical, chemical or enzymatic.
  • Chemical hydrolysis of e.g. lignocellulosic material can normally be achieved in a manner known per se by treatment with an acid, such as treatment with dilute (e.g. 2-10% w/w, typically 4-7% w/w) aqueous sulphuric acid, at a temperature in the range of about 100- 150°C, e.g. around 120°C, for a period of 5-15 minutes, such as 5-10 minutes. Treatment with ca. 4% w/w sulphuric acid for 5-10 minutes at ca. 120°C is often very suitable.
  • an acid such as treatment with dilute (e.g. 2-10% w/w, typically 4-7% w/w) aqueous sulphuric acid, at a temperature in the range of about 100- 150°C, e.g. around 120°C, for a period of 5-15 minutes, such as 5-10 minutes.
  • Treatment with ca. 4% w/w sulphuric acid for 5-10 minutes at ca. 120°C
  • the release of the carbohydrate-containing material as well as the provision of accessible sugars from the carbohydrate-containing material can be obtained by enzymatic treatment.
  • cellulosic materials cellulases and hemicellulases may be used. Suitable microorganisms may be used to produce such enzymes.
  • the sterilised wastewater can serve as water phase during the process.
  • the grain Prior to enzymatic hydrolysis and fermentation the grain must be opened by physical methods to release the starch.
  • the two best-known and most widely used processes in starchy materials are wet milling and dry milling.
  • wet milling the corn is first steeped in a solution of water and sulphur dioxide for 24-48 hours, at a temperature of around 52°C, and then passed through mills to loosen the germ and the hull fibres.
  • the dry milling process the grain is first broken up as small particle size as possible in order to facilitate subsequent penetration of water in the following cooking process described below in which waste water can be used.
  • the milled starchy material Before fermentation, the milled starchy material must be processed (“saccharified”), to convert the starch into fermentable sugars by distillery yeast (Baker's Yeast). This can be done by use of starch hydrolysing enzymes - amylases. In its natural state, starch exists as compact crystalline granules, which are resistant to enzymatic attack. To enable the enzymes to operate more efficiently, the starch molecules are brought into solution e.g. by using wastewater by a heat treatment.
  • the raw material is first prepared as a slurry of found meal in wastewater.
  • a small quantity of ⁇ - amylase is added to reduce the viscosity of the slurry during the following cooking procedure.
  • the slurry is then cooked at 130-160°C.
  • the starch Once the starch has gelatinised the mash is cooled to 80-90°C at which temperature the -amylase can be added to produce rapid liquefaction.
  • the amyloglycosidase can be added together with yeast to carry out the hydrolysis and fermentation.
  • SHF separate hydrolysis and fermentation.
  • SSF separate hydrolysis and fermentation
  • the present invention relates to a fermentation medium comprising wastewater and to the use of such a fermentation medium for the production of a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • the fermentation medium and/or the use of such a fermentation medium is not for the production of methane.
  • the fermentation medium may be useful in the production of biomass of a selected microorganism such as bacteria, fungi or yeast, including S. cerevisiae. It will be understood that the above-described embodiments of the fermentation process of the invention also relate to the fermentation medium of the present invention and its use in fermentation processes.
  • the fermentation medium comprises wastewater which is obtained from sewage selected from the group consisting of municipal sewage, household waste, slaughterhouse waste, human waste, animal waste and/or industrial waste such as waste from the food, feed and pharmaceutical industry such as cell biomass including genmodified cell biomass.
  • the fermentation medium comprises a biomass material which may be a carbohydrate-containing material as described above.
  • the biomass material may be, as discussed above, pretreated by thermal treatment, wet oxidation, steam explosion, dilute sulphuric acidic or other relevant acids, alkaline solutions, organic solvents or any combination hereof before being subjected to fermentation.
  • the pretreated or untreated biomass is hydrolysed by enzymatic treatment or chemical hydrolysis, as discussed above.
  • the fermentation medium is one wherein the ratio between the above discussed carbohydrate-containing material and the wastewater is in the range of 1:99-1 : 1, preferably in the range of 1:49-1:2, including the range of 1:9-1:4.
  • the wastewater may be treated prior to adding the biomass material, such a treatment may be a thermal treatment, fermentation, enzymatic treatment, acid hydrolysis, wet oxidation, filtration or any combination thereof.
  • the wastewater may be supplemented with at least one nutrient compound selected from the group consisting of nitrogen, phosphor, magnesium, zinc, manganese, cobalt, copper, calcium, iron, molybde- num, boron, a compound containing any of such elements and any mixture thereof.
  • the fermentation medium according to the present invention may be used in the production of a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • Fig. 1 shows a schematic view of the process according to the present invention
  • Fig. 2 shows the ethanol production at two enzyme loadings during fermentation of paper pulp in wastewater derived from thermally treated and anaerobic digested sewage.
  • a concentration of cellulose of 7 FPU/g dry matter (triangles) and 10 FPU/g of dry matter (crosses);
  • Fig. 3 shows the cellulose conversion at two enzyme loadings during fermentation of paper pulp to ethanol in wastewater derived from thermally treated and anaerobic digested sewage.
  • a concentration of cellulose 7 FPU/g dry matter (triangles) and 10 FPU/g of dry matter (crosses);
  • Fig. 4 shows the production of ethanol during fermentation of wet oxidised wheat straw and paper pulp, respectively, in wastewater (derived from thermally treatment and anaerobic digestion of sewage) with and without addition of urea.
  • wastewater derived from thermally treatment and anaerobic digestion of sewage
  • a paper pulp (circles), paper pulp and urea (star), wheat straw (crosses and dashed line) and wheat straw and urea (squares);
  • Fig. 5 shows the conversion of cellulose during fermentation of paper pulp and wet oxidised wheat straw in wastewater (derived from thermally treatment and anaerobic diges- tion of sewage) with and without urea addition.
  • wastewater derived from thermally treatment and anaerobic diges- tion of sewage
  • a paper pulp (circles), paper pulp and urea (star), wheat straw (crosses and dashed line) and wheat straw and urea (squares);
  • Fig. 6 shows the conversion of cellulose during ethanol fermentation using wastewater derived from slaughterhouse waste (MBF).
  • MBF treated by wet oxidation WO-MBF, triangles
  • MBF treated by wet oxidation followed by anaerobic digestion WO-AD-MBF, diamonds
  • MBF treated wet oxidation followed by anaerobic digestion and addition of urea WO-AD-MBF+urea, squares.
  • Wet oxidised wheat straw was used as biomass;
  • Fig.7 shows the production of ethanol during fermentations using wastewater from slaughterhouse waste, i.e. meat and bone flour (MBF).
  • MBF treated by wet oxidation WO-MBF, triangles
  • MBF treated by wet oxidation followed by anaerobic digestion WO-AD-MBF, diamonds
  • MBF treated wet oxidation followed by anaerobic digestion and addition of urea WO-AD-MBF+urea, squares.
  • Wet oxidised wheat straw was used as biomass;
  • Fig 8 shows the conversion of cellulose during ethanol fermentation using wastewater from manure.
  • manure treated by wet oxidation WO- manure, triangles
  • manure treated by wet oxidation followed by anaerobic digestion WO- AD-manure, diamonds
  • manure treated wet oxidation followed by anaerobic digestion and addition of urea WO-AD-manure+urea, squares.
  • Wet oxidised wheat straw was used as biomass;
  • Fig. 9 shows the production of ethanol during fermentation using wastewater from manure.
  • manure treated by wet oxidation WO-manure, triangles
  • manure treated by wet oxidation followed by anaerobic digestion WO-AD-ma- nure, diamonds
  • manure treated wet oxidation followed by anaerobic digestion and addition of urea WO-AD-manure+urea, squares.
  • Wet oxidised wheat straw was used as biomass; and
  • Fig. 10 shows the production of ethanol during fermentation using household waste (HW).
  • household waste treated by wet oxidation WO-HW, diamonds
  • household waste treated by wet oxidation and addition of urea WO- HW+urea, squares.
  • This example illustrates the effect of wastewater in the fermentation of Saccharomyces cerevisiae (Baker's yeast) in order to produce ethanol.
  • Ethanol has in recent years received attention as a potential replacement for or supplement to petroleum-derived liquid hydro- carbon products.
  • ethanol for use in the food industry is produced from molasses and starch, but now where ethanol may be used as a fuel (not to consume), sewage and/or wastewater have now surprisingly shown to be potential candidates for the supply of nutrients and water in the ethanol fermentation.
  • the wastewater derived from sewage was subjected to thermal hydrolysis at 180°C for one hour followed by anaerobic microbial digestion and filtration as indicated in Figure 1 (flow- diagram).
  • the chemical data for the wastewater is shown in Table 1.1.
  • the cellulose content is estimate to 93% (w/w) and 7% inorganic material (mainly kaolin).
  • the suspension was cooled on ice (to room temperature) and 0.8 g freeze dried Baker's yeast and 1 ml 24% urea (corresponding to 1000 mg/l) was added together with another portion of enzymes making the total enzyme loadings in the two experiments at 7 and 10 10 FPU/g DM, respectively.
  • Paper pulp (as used in Example 1) derived from processing wastepaper for reuse.
  • the cellulose content is estimate to 93% (w/w) and 7% inorganic material (mainly kaolin).
  • Pre-treated wheat straw Wheat straw was treated by wet oxidation using 60 g in one litre of wastewater heated to 195°C for 10 minutes at initial 12 bars of oxygen pressure (giving a total pressure of 20 bars); after the treatment the slurry was cooled and filtered and washed with 200 ml of water.
  • the composition of the wet oxidised wheat straw is seen in Table 2.1: Table 2.1: Chemical composition of pre-treated wheat straw and untreated wheat straw.
  • This example illustrates that two different cellulosic materials, wheat straw and paper pulp respectively, each being mixed with wastewater derived from thermally treated and an- aerobically digested sewage can serve as the only substrates without addition of urea.
  • the experiments were carried out as outlined in Example 1, and compared with parallel experiments without addition of urea.
  • the enzyme loading was in all experiments 7 FPU/g DM.
  • the ethanol production ( Figure 4) was highest using paper pulp as a substrate compared to wet oxidised wheat straw due to the higher cellulose content however the conver- sion of the cellulose in percentage was similar for the two substrates ( Figure 5).
  • the fermentation and ethanol productivity showed to be independent of urea addition.
  • the ethanol yield measured after fermentation was in accordance with the calculated ethanol yields based on the C0 2 production with a small difference (5%) due to a minor yeast production in the initial fermentation phase facilitated by the amount of oxygen present in the flask head space.
  • Table 2.2 shows the ethanol produced during fermentation of wet oxidised wheat straw and paper pulp respectively in wastewater (derived from thermally treatment and anaerobic digestion) with and without addition of urea also as indicated in Figure 4.
  • Table 2.2 Ethanol yield measured by HPLC.
  • This example illustrates that wastewater derived from wet oxidised slaughterhouse waste, i.e. meat and bone flour, can be used in ethanol fermentation.
  • Wastewater The wastewater used in this example was obtained from wet oxidised slaughterhouse waste, i.e. meat and bone flour (MBF).
  • MBF meat and bone flour
  • the content of MBF is shown in Table 3.1.
  • Wet oxidation (WO) of meat and bone flour was carried out in a 2 L loop autoclave treating 1 litre of water containing 100 g MBF at 200°C for 15 minutes with 12 bars of 0 2 in the headspace of the reactor of one litre. After the treatment the suspension was filtered into a liquid and a solid fraction.
  • the ash content of MBF before wet oxidation was 41.6% and after wet oxidation 83.4% (corresponding to 46.2 g mainly inorganic particles) showing that a significant amount of the organic components (protein) were dissolved during the wet oxidation process.
  • the liquid fraction was analysed for Na + , K + and NH 4 + (Table 3.2).
  • the anaerobic digestion was performed in bluecap-flasks with yeast locks by adding methanogens to the liquid and leaving it on a lab shaker at anaerobic conditions for 6 days after which the organic carbon fraction was reduced by 2.6 g/L measure by weight loss.
  • the anaerobic digested sample was then sterilised to stop biological activity.
  • WO wheat straw was used as biomass material in this example.
  • WO wheat straw was prepared by wet oxidation of 60 g of wheat straw in one litre of water supplied with 2 g Na 2 C0 3 at 195°C, for 12 minutes with 12 bars of oxygen added to the headspace of the reactor volume of one litre. After the treatment the cooled suspen- sion was filtered and the filter cake was analysed (Table 3.3) for its content of cellulose, hemicellulose, lignin and ash. The filter cake was stored in a climate chamber before used as substrate in the fermentations.
  • Simultanous saccharification and fermentation also referred to the mandatory fermentation, was carried out in blue cap flasks with yeast locks.
  • the dry matter content of wet oxidised wheat straw was 8%.
  • wet oxidised MBF and/or wet oxidised and anaerobically digested MBF can serve as the only substrate in an ethanol fermentation without addition of additional nutrients such as urea.
  • wastewater derived from MBF can be an essential component in a fermentation medium the production of a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • This example illustrates that wastewater derived from wet oxidised manure can be used in ethanol fermentation.
  • the wastewater used in this example was obtained from digested manure obtained from a biogas plant containing 3% solid matter followed by a wet oxidiation.
  • Wet oxidation of manure (see composition in Table 4.2) was carried out in a 2 L loop autoclave treating 1 litre of manure at 200°C, 15 minutes with 12 bars of 0 2 supplied to the headspace of the re- actor of one litre. After the treatment no more solid was present and also the odour was vanished.
  • the liquid fraction was analysed for Na + , K + and NH + (Table 4.1).
  • the anaerobic digestion was performed in bluecap-flasks with yeast locks by adding methanogens to the liquid and leaving it on a lab shaker at an- aerobic conditions for 6 days after which the organic carbon fraction was reduced by 0.6 g/L measure by weight loss.
  • the anaerobic digested sample was then sterilised to stop biological activity.
  • WO wheat straw was used as biomass material in this example.
  • WO wheat straw was prepared by wet oxidation of 60 g of wheat straw in one litre of water supplied with 2 g Na 2 C0 3 at 195°C, for 12 minutes with 12 bars of oxygen added to the headspace of the reactor volume of one litre. After the treatment the cooled suspension was filtered and the filter cake was analysed (Table 4.2) for its content of cellulose, hemicellulose, lignin and ash. The filter cake was stored in a climate chamber before used as substrate in the fermentations.
  • FIG. 8 The cellulose conversion during ethanol fermentation and the production of ethanol are shown in Figures 8 and 9, respectively.
  • This example illustrates that wastewater derived from manure can be used as a source for water and nutrients in an ethanol fermentation using wet oxidised wheat straw as a carbon source.
  • the highest ethanol yield was produced using wastewater derived from manure which only had been treated by wet oxidation, suggesting that an anaerobic digestion of the wastewater is not needed.
  • the fermentation was completed after 170 hours reaching a level of 23 g/l. This corresponded to about 98% conversion of the cellulose.
  • wet oxidised manure and/or wet oxidised and anaerobically digested manure can serve as the only substrate in an ethanol fermentation without addition of nutrients such as urea.
  • wastewater derived from manure can be an essential component in a fermentation medium the production of a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins and enzymes.
  • This example illustrates that wastewater as well as the glucan containing solids derived from wet oxidised household waste can be used in ethanol fermentation.
  • Household waste was collected from a municipal waste treatment plant in Frederikssund (Denmark). It consisted of source-sorted kitchen waste (Table 5.1) shred- ded to ⁇ 1 mm and enriched with wheat straw (8%) for stabilization of the waste. 60 g MSW in one liter of water was treated by wet oxidation for 10 minutes at 195°C, supplemented with 2 g of Na 2 C0 3 and 12 bars of oxygen added to the headspace of the reactor volume of one liter. After the treatment the reactor was cooled and the suspension filtered. Table 5.1 Chemical composition of raw and treated household waste
  • Both the filter cake and filtrate were used in the fermentation as biomass material and wastewater, respectively. Fermentations were carried out with and without addition of urea. Liquefaction was obtained using 5 FPU/g DM (celluclast and Novozym 188, 5: 1) at 50°C for 24 hours after which the suspensions were cooled and supplied with more enzymes, 20 FPU/g DM (celluclast and Novozym 188, 5: 1) together with 0.3% w/v dry Bak- ers yeast. The fermentations were carried out at 32°C and the ethanol production measured by weight loss due to C0 2 and ethanol production (g/L), as described in Example 1.
  • FIG. 10 The production of ethanol is shown in Figure 10.
  • This example illustrates that wastewater as well as the solid fraction derived from household waste (HW) can be used as a source for water and nutrients and substrate in an ethanol fermentation and that the solid phase of the waste contains sufficient carbon for a fermentation.
  • HW household waste
  • the fermentation was completed after 250 hours reaching a level of 23 g/l.
  • the final ethanol yield corresponded to a glucan conversion of 81-82%.
  • wastewater derived from household waste can be an essential component in a fermentation medium the production of a fermentation product selected from the group consisting of ethanol, lactic acid, acetate, propionate, butyrate, formate, hydrogen, H 2 , C0 2 , vitamins, antibiotics, amino acids, colours, proteins, enzymes and cell biomass.
  • Thygesen A., Thomsen, A.B., Schmidt, A.S., J ⁇ rgensen, H., Ahring, B.K. & Olsson, L. 2003. Production of cellulose and hemicellulose-degrading enzymes by filamentous fungi cultivated on wet oxidised wheat straw. Enzyme and Microbial Technology 32:606-615.

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Abstract

La présente invention se rapporte à un milieu de fermentation comprenant des eaux usées et à un procédé de production des produits de fermentation, notamment de l'acide lactique, à un produit de carburant, par exemple de l'éthanol ou une biomasse cellulaire au moyen d'eaux usées d'égouts, à savoir les égouts municipaux, les déchets d'abattoirs, les déchets domestiques ou le fumier. Le procédé consiste à : (i) fournir les eaux usées, (ii) ajouter une matière biomasse aux eaux usées de l'étape (i) afin d'obtenir une suspension de biomasse, et (iii) soumettre la suspension de l'étape (ii) à un procédé de fermentation afin d'obtenir un produit de fermentation, et (iv) séparer le produit de fermentation obtenu à l'étape (iii).
PCT/DK2004/000385 2003-06-06 2004-06-04 Milieu de fermentation comprenant des eaux usees et son utilisation Ceased WO2004108609A1 (fr)

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CN100343187C (zh) * 2005-05-24 2007-10-17 李庚承 生产vb12的工业废水的资源化处理工艺及其专用废水处理机
WO2009142784A1 (fr) 2008-05-21 2009-11-26 Applied Cleantech Inc. Procédés et systèmes de fabrication de matière première à partir d’eaux usées et fabrication de produit grâce à ces derniers
ITMI20091550A1 (it) * 2009-09-09 2011-03-10 Eni Spa Procedimento per la produzione di bio- olio da rifiuti solidi urbani
US7968760B2 (en) 2007-03-16 2011-06-28 Ch2M Hill, Inc. Treatment of particulate biodegradable organic waste by thermal hydrolysis using condensate recycle
CN102174602A (zh) * 2011-03-07 2011-09-07 南京林业大学 一种利用生物质发酵生产l-乳酸的方法
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ITMI20100646A1 (it) * 2010-04-15 2011-10-16 Eni Spa Procedimento per la produzione di bio-olio da rifiuti solidi urbani
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WO2014113615A1 (fr) * 2013-01-16 2014-07-24 Clean-Vantage Llc Oxydation humide de biomasse
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