Method of controlling fermentation of a co-containing substrate

Abstract

This invention relates to a method for controlling the fermentation of CO-containing substrates. A method for stabilizing the fermentation of CO-containing substrates and increasing ethanol production includes providing culture medium components in amounts required by microorganisms during fermentation. The method includes determining the potassium concentration in the fermentation medium and providing a first culture medium and a second culture medium to the fermentation, the first culture medium being provided at a rate that effectively maintains potassium in the fermentation medium at a concentration of about 20 to about 200 mg / L until a target cell density is reached.

Description

[0001] This application is a divisional application of Chinese Patent Application 201580043232.8, "Method for controlling fermentation of CO-containing substrates", filed on July 24, 2015.

[0002] This application claims the benefit of U.S. Provisional Application 62 / 036,239, filed August 12, 2014, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention provides a method for fermenting a CO-containing substrate. More specifically, the method includes determining the concentration of nutrients in the fermentation medium and maintaining those concentrations within a specified range. The method also includes maintaining a nutrient concentration effective for providing 10 g total alcohols / (L·day) or greater. Background Technology

[0004] Fermentation takes place in defined liquid media. These media typically include a variety of macronutrient and micronutrient sources important for improving fermentation performance. Media used with uncommon substrates, such as gaseous substrates, require well-defined media to optimize performance. Anaerobic fermentation also requires well-defined media.

[0005] Anaerobic microorganisms can produce ethanol from carbon monoxide (CO) through the fermentation of gaseous substrates. Anaerobic microorganisms can be used to ferment ethanol from Clostridium species (…). Clostridium Fermentation produces ethanol and other useful products. For example, U.S. Patent 5,173,429 describes *Clostridium yangensis* (…). Clostridium ljungdahlii ATCC No. 49587, an anaerobic microorganism that produces ethanol and acetate from syngas. US Patent 5,807,722 describes the use of *Clostridium yangrensis* (…). Clostridium ljungdahlii ATCC No. 55380 describes a method and apparatus for converting waste gas into organic acids and alcohols. US Patent 6,136,577 describes the use of Clostridium jansii (…). Clostridium ljungdahlii ATCC Nos. 55988 and 55989: Methods and apparatus for converting waste gas into ethanol.

[0006] U.S. Patent 7,285,402 describes known culture media for anaerobic fermentation of gaseous substrates to produce ethanol. The various components and concentrations in the culture medium effectively provide high levels of ethanol production. Providing certain culture medium components required by the microorganisms at different times during fermentation can provide improved ethanol production and a more stable fermentation process. Summary of the Invention

[0007] A method for stabilizing the fermentation of a CO2-containing substrate and increasing ethanol production includes providing culture medium components in the amount required by the microorganisms for fermentation. More specifically, the method for fermenting a CO2-containing substrate includes providing a CO2-containing substrate to the fermentation medium and allowing the CO2-containing substrate to ferment. The method also includes monitoring nutrient concentrations in the fermentation medium using techniques selected from: ion chromatography, inductively coupled plasma atomic absorption spectrometry, ion-selective electrodes, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof.

[0008] In another aspect, a method for fermenting a CO-containing substrate includes providing a CO-containing substrate to the fermenter and fermenting the CO-containing substrate; determining the concentration of potassium in the fermentation medium; and providing a first medium and a second medium to the fermenter, the first medium being provided at a rate that effectively maintains about 20 to about 200 mg / L of potassium in the fermentation medium until a target cell density is reached. Invention Details The following description should not be construed as limiting, but is intended only to illustrate the general principles of exemplary embodiments. The scope of the invention should be defined by the claims.

[0010] definition Unless otherwise defined, the following terms, as used throughout this disclosure, are defined as follows and may include the singular or plural forms of the following definitions: The term “about”, which modifies any quantity, refers to variations in quantity encountered under real-world conditions, such as in a laboratory, pilot plant, or production facility. For example, when modified by “about,” the quantity of an ingredient or measurement used in a mixture or quantity includes variations and extents typically considered when tested under experimental conditions in a production facility or laboratory. For example, when modified by “about,” the quantity of a product component includes variations between batches tested multiple times in a facility or laboratory and variations inherent to the analytical method. Whether or not modified by “about,” the quantity includes equivalent quantities of those quantities. Any quantity stated herein and modified by “about” can also be used in this disclosure as a quantity not modified by “about.”

[0011] As used herein, "carbon-containing materials" refers to carbon-rich materials, such as coal and petrochemical products. However, in this specification, carbon-containing materials include any carbon material, whether in a solid, liquid, gaseous, or plasma state. Among the many items that can be considered carbon-containing materials, this disclosure anticipates: carbon-containing materials, carbon-containing liquid products, carbon-containing industrial liquid recyclables, carbon-containing municipal solid waste (MSW or MSW), carbon-containing municipal waste, carbon-containing agricultural materials, carbon-containing forestry materials, carbon-containing wood waste, carbon-containing building materials, carbon-containing plant materials, carbon-containing industrial waste, carbon-containing fermentation waste, carbon-containing petrochemical byproducts, carbon-containing alcohol production byproducts, carbon-containing coal, tires, plastics, waste plastics, coke oven tar, soft fibers, lignin, black liquor, polymers, waste polymers, polyethylene terephthalate (PETA), polystyrene (PS), sewer sludge, animal waste, crop residues, energy crops, forestry processing residues, wood processing residues, livestock waste, poultry waste, food processing residues, fermentation process waste, ethanol byproducts, waste particles, waste microorganisms, and combinations thereof.

[0012] The term "fibersoft" (or "fibrosoft" or "fibrousoft") refers to a carbon-containing material produced by softening and concentrating various substances; in one instance, the carbon-containing material is produced via vapor pressure heat treatment of various substances. In another instance, soft fibers may include fibrous paste-like materials produced by vapor pressure heat treatment of municipal, industrial, commercial, and medical waste.

[0013] The terms “municipal solid waste” or “MSW” or “msw” refer to waste that may include household, commercial, industrial and / or residual waste.

[0014] The term "syngas" or "synthetic gas" refers to a gaseous mixture containing varying amounts of carbon monoxide and hydrogen. Examples of production methods include natural gas or hydrocarbon steam reforming to produce hydrogen, coal gasification, and some types of waste-to-energy gasification plants. The name derives from their use as intermediates in the production of synthetic natural gas (SNG) and its use in the production of ammonia or methanol. Syngas is combustible and is commonly used as a fuel source or as an intermediate in the production of other chemicals.

[0015] "Ton" or "ton" refers to the US short ton, which is approximately 907.2 kg (2000 lbs).

[0016] Productivity used in this paper is denoted as STY. In this respect, ethanol productivity can be expressed as STY (space-time productivity, expressed as g ethanol / (L·day)). Detailed Implementation

[0017] The present invention also relates to the following embodiments: 1. A method for controlling the fermentation of a CO-containing substrate, the method comprising: Providing a CO-containing substrate to fermentation and causing the CO-containing substrate to ferment; and Nutrient concentrations in fermentation media can be monitored using techniques selected from the following: ion chromatography, inductively coupled plasma atomic absorption spectrometry, ion-selective electrode, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof.

[0018] 2. The method of implementation plan 1, wherein the monitored nutrients are selected from K, Mg, and PO4. -3 Fe, Zn, Ni, Co and their mixtures.

[0019] 3. The method of implementation plan 2, wherein the nutrients being monitored are K and PO4. -3 Mg and its mixtures.

[0020] 4. The method of implementation scheme 1, wherein the technique used to monitor nutrient concentration is ion chromatography.

[0021] 5. The method of implementation 1, wherein the method provides a target cell density of about 2 to about 30 g / L.

[0022] 6. The method of embodiment 5, wherein the fermentation medium comprises first and second nutrient-containing mediums.

[0023] 7. The method of embodiment 6, wherein the first culture medium is provided to fermentation at a rate that effectively maintains a potassium concentration of about 20 to about 200 mg / L in the culture medium until the target cell density is reached.

[0024] 8. The method of embodiment 7, wherein after the target cell density is reached, the first culture medium is provided to fermentation at a rate that effectively maintains a potassium concentration of about 20 to about 160 mg / L in the culture medium.

[0025] 9. The method of embodiment 6, wherein the first culture medium is used to effectively maintain a concentration of about 10 to about 120 mg / L PO4. -3 The concentration is supplied at a rate that continues to ferment until the target cell density is reached.

[0026] 10. The method of embodiment 9, wherein after the target cell density is reached, the first culture medium is used to effectively maintain a concentration of approximately 10 to approximately 100 mg / L PO4. -3 The concentration rate is provided to the fermentation process.

[0027] 11. The method of embodiment 6, wherein the first culture medium is provided to fermentation at a rate that effectively maintains a concentration of about 1 to about 6 mg / L mg in the culture medium until the target cell density is reached.

[0028] 12. The method of embodiment 11, wherein after the target cell density is reached, the first culture medium is provided to fermentation at a rate that effectively maintains a Mg concentration of about 1 to about 4 mg / L in the culture medium.

[0029] 13. The method of embodiment 6, wherein the second culture medium is provided to fermentation at a rate that effectively maintains a volumetric flow rate ratio of about 15:1 to about 2:1 between the first and second culture media.

[0030] 14. The method of embodiment 6, wherein the first culture medium comprises a mixture selected from K, Mg, Fe, PO4. -3 Elements including Zn, Co, Ni and their mixtures.

[0031] 15. The method of embodiment 6, wherein the second culture medium comprises an element selected from W, Se, and mixtures thereof.

[0032] 16. The method of embodiment 1, wherein the CO-containing substrate has a CO / CO2 molar ratio of at least about 0.75.

[0033] 17. The method of embodiment 1, wherein the CO-containing substrate has about 20 to about 100% molar CO.

[0034] 18. The method of embodiment 1, wherein the pH of the fermentation medium is maintained at a level of about 3.5 to about 5.0.

[0035] 19. The method of implementation scheme 18, wherein the pH is maintained by adding ammonium hydroxide.

[0036] 20. The method of embodiment 1, the method further comprising maintaining an H2S concentration of about 20 to about 500 ppm H2S in the fermentation exhaust gas.

[0037] 21. The method of embodiment 20, wherein a H2S concentration of about 20 to about 500 ppm is maintained in the fermentation exhaust gas by adding NaHS to the fermentation medium.

[0038] 22. The method of implementation scheme 1, wherein the fermentation includes acetic acid-producing bacteria.

[0039] 23. The method of embodiment 22, wherein the acetic acid-producing bacteria is selected from Kavulcanii (Acetobacter acetogenes). Acetogenium I waved ), moist anaerobic acetic acid bacteria ( Acetoanaerobic record Acetobacter wuerii (), Acetobacterium wood ), Bacillus bacitratus ( Alkalibaculum bacchi CP11 (ATCC BAA-1772), Propionibacterium brevicornu ( Blautia products ), methylbutyric acid bacteria ( Butyribacterium methylotrophicum), underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneus ), Pacific underground thermophilic anaerobic bacteria ( Caldanaerobacter underground peaceful ), hydrogen-producing carboxyl thermophilic bacteria ( Carboxydothermus hydrogenoformans Acetic acid bacteria () Clostridium aceticum Clostridium acetonebutanol ( Clostridium acetobutylicum Clostridium acetonebutanol ( Clostridium acetobutylicum P262 (DSM 19630 from DSMZ, Germany), self-produced Clostridium ethanoliferum ( Clostridium autoethanogenum (DSM 19630 of the German DSMZ) 、 Self-produced Clostridium ethanol ( Clostridium autoethanogen (DSM 10061 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogen (DSM 23693 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogen (DSM 24138 from DSMZ, Germany), Clostridium carboxylate ( Clostridium carboxydivorans ) P7 (ATCC PTA-7827), Clostridium krill ( Clostridium coskatii (ATCC PTA-10522), Clostridium decoctioni ( Clostridium drakei Clostridium yangii ( Clostridium ljungdahlii ) PETC (ATCC 49587), Clostridium yangii ( Clostridium ljungdahlii ) ERI2 (ATCC 55380), Clostridium yangii ( Clostridium ljungdahlii ) C-01 (ATCC 55988), Clostridium yangii ( Clostridium ljungdahlii ) O-52 (ATCC 55889), Clostridium macrocarpa ( Clostridium macrophyllum ), Clostridium pasteurellum ( Clostridium pasteurianum (DSM 525 from DSMZ in Germany), Clostridium laurylidum ( Clostridium ragsdali ) P11 (ATCC BAA-622), Clostridium paederi ( Clostridium scatologist ), Clostridium thermophilum ( Clostridium thermoaceticum ), Clostridium Tunneloni ( Clostridium ultunense ), Kuhl's desulfurized enterobacteria ( Desulfotomaculum kuznetsovii ), Eubacterium mucilaginosa ( Eubacterium slime ), sulfur-reducing bacteria ( Geobacter sulfurreducens ), Methanophora acetophenones ( Methanosarcina acetivorans ), Pasteurella multocida ( Methanosarcina barkeri ), thermoacetic acid murine bacteria ( Morrella thermoacetica ), thermoautotrophic Murraya ( Morella thermoautotrophic ), Fanny acetobacterium ( Oxobacter pfennigii ), producing Peptostreptococcus ( Peptostreptococcus productus ), producing rumen cocci ( Ruminococcus productus ), Kaiwu thermophilic anaerobic bacteria ( Thermoanaerobacter kivui ) and their mixtures.

[0040] 24. A method for fermenting a CO-containing substrate, the method comprising: Provide CO-containing substrates to fermentation and enable the CO-containing substrates to ferment; Determine the potassium concentration in the fermentation medium; and A first culture medium and a second culture medium are provided to the fermentation, the first culture medium being provided at a rate of approximately 20 to approximately 200 mg / L potassium in the fermentation medium until the target cell density is reached.

[0041] 25. The method of embodiment 24, wherein after the target cell density is reached, the first culture medium is provided to fermentation at a rate that effectively maintains a potassium concentration of about 20 to about 160 mg / L in the culture medium.

[0042] 26. The method of embodiment 24, wherein the CO-containing substrate has a CO / CO2 molar ratio of at least about 0.75.

[0043] 27. The method of embodiment 24, wherein the CO-containing substrate has about 20 to about 100% molar CO.

[0044] 28. The method of implementation scheme 24, wherein the concentration of potassium in the culture medium is determined by a method selected from the following: ion chromatography, inductively coupled plasma atomic absorption spectrometry, ion-selective electrode, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof.

[0045] 29. The method of embodiment 24, wherein the target cell density is about 2 to about 30 g / L.

[0046] 30. The method of embodiment 24, wherein the second culture medium is provided to fermentation at a rate that effectively maintains a volumetric flow rate ratio of about 15:1 to about 2:1 between the first and second culture media.

[0047] 31. The method of embodiment 24, wherein the first culture medium comprises a mixture selected from K, Mg, Fe, PO4. -3 Elements including Zn, Co, Ni and their mixtures.

[0048] 32. The method of embodiment 24, wherein the second culture medium comprises an element selected from W, Se and mixtures thereof.

[0049] 33. The method of embodiment 24, wherein the pH of the fermentation medium is maintained at a level of about 3.5 to about 5.0.

[0050] 34. The method of implementation scheme 24, wherein the pH is maintained by adding ammonium hydroxide.

[0051] 35. The method of embodiment 24, the method further comprising maintaining an H2S concentration of about 20 to about 500 ppm H2S in the fermentation exhaust gas.

[0052] 36. The method of embodiment 35, wherein a H2S concentration of about 20 to about 500 ppm is maintained in the fermentation exhaust gas by adding NaHS to the fermentation medium.

[0053] 37. The method of embodiment 24, wherein the fermentation includes acetic acid-producing bacteria.

[0054] 38. The method of embodiment 37, wherein the acetic acid-producing bacteria is selected from Kavulcanii (Acetobacter acetogenes). Acetogenium I waved ), moist anaerobic acetic acid bacteria ( Acetoanaerobic record Acetobacter wuerii (), Acetobacterium wood ), Bacillus bacitratus ( Alkalibaculum bacchi CP11 (ATCC BAA-1772), Propionibacterium brevicornu ( Blautia products ), methylbutyric acid bacteria ( Butyribacterium methylotrophicum ), underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneus ), Pacific underground thermophilic anaerobic bacteria ( Caldanaerobacter underground peaceful ), hydrogen-producing carboxyl thermophilic bacteria ( Carboxydothermus hydrogenoformans Acetic acid bacteria () Clostridium aceticum Clostridium acetonebutanol ( Clostridium acetobutylicum Clostridium acetonebutanol ( Clostridium acetobutylicum P262 (DSM 19630 from DSMZ, Germany), self-produced Clostridium ethanoliferum ( Clostridium autoethanogenum (DSM 19630 of the German DSMZ) 、 Self-produced Clostridium ethanol ( Clostridium autoethanogen (DSM 10061 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogen (DSM 23693 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogen (DSM 24138 from DSMZ, Germany), Clostridium carboxylate ( Clostridium carboxydivorans ) P7 (ATCC PTA-7827), Clostridium krill ( Clostridium coskatii(ATCC PTA-10522), Clostridium decoctioni ( Clostridium drakei Clostridium yangii ( Clostridium ljungdahlii ) PETC (ATCC 49587), Clostridium yangii ( Clostridium ljungdahlii ) ERI2 (ATCC 55380), Clostridium yangii ( Clostridium ljungdahlii ) C-01 (ATCC 55988), Clostridium yangii ( Clostridium ljungdahlii ) O-52 (ATCC 55889), Clostridium macrocarpa ( Clostridium macrophyllum ), Clostridium pasteurellum ( Clostridium pasteurianum (DSM 525 from DSMZ in Germany), Clostridium laurylidum ( Clostridium ragsdali ) P11 (ATCC BAA-622), Clostridium paederi ( Clostridium scatologist ), Clostridium thermophilum ( Clostridium thermoaceticum ), Clostridium Tunneloni ( Clostridium ultunense ), Kuhl's desulfurized enterobacteria ( Desulfotomaculum kuznetsovii ), Eubacterium mucilaginosa ( Eubacterium slime ), sulfur-reducing bacteria ( Geobacter sulfurreducens ), Methanophora acetophenones ( Methanosarcina acetivorans ), Pasteurella multocida ( Methanosarcina barkeri ), thermoacetic acid murine bacteria ( Morrella thermoacetica ), thermoautotrophic Murraya ( Morella thermoautotrophic ), Fanny acetobacterium ( Oxobacter pfennigii ), producing Peptostreptococcus ( Peptostreptococcus productus ), producing rumen cocci ( Ruminococcus productus ), Kaiwu thermophilic anaerobic bacteria ( Thermoanaerobacter kivui ) and their mixtures.

[0055] 39. The method of embodiment 24, wherein the method is effective for providing 10 g total alcohol / (L·day) or greater STY.

[0056] 40. The method of embodiment 24, the method further comprising effectively maintaining about 10 to about 120 mg / L PO4 in the culture medium. -3 The concentration rate provides the first culture medium to the fermentation process until the target cell density is reached.

[0057] 41. The method of embodiment 24, the method further comprising providing a first culture medium to fermentation at a rate that effectively maintains a concentration of about 1 to about 6 mg / L mg in the culture medium until a target cell density is reached.

[0058] CO-containing substrate CO-containing substrates can include any gas containing CO. In this respect, CO-containing gases can include syngas, industrial gases, and mixtures thereof.

[0059] Syngas can be provided from any known source. On one hand, syngas can be derived from the gasification of carbonaceous materials. Gasification involves the partial combustion of biomass in a limited supply of oxygen. The resulting gas primarily comprises CO and H2. On one hand, the syngas will contain at least about 10 mol% CO, on another hand, at least about 20 mol% CO, on another hand, about 10–about 100 mol% CO, on another hand, about 20–about 100 mol% CO, on another hand, about 30–about 90 mol% CO, on another hand, about 40–about 80 mol% CO, and on another hand, about 50–about 70 mol% CO. Examples of suitable gasification methods and apparatus are provided in U.S. Serial Nos. 61 / 516,667, 61 / 516,704 and 61 / 516,646, filed April 6, 2011, and U.S. Serial Nos. 13 / 427,144, 13 / 427,193 and 13 / 427,247, filed March 22, 2012, all of which are incorporated herein by reference.

[0060] On the other hand, the method is applicable to supporting the production of alcohols from gaseous substrates (e.g., industrial flue gas containing high volumes of CO). In some aspects, the CO-containing gas originates from carbonaceous waste (e.g., industrial exhaust gas) or from the gasification of other wastes. Therefore, the method represents an effective approach for capturing carbon that would otherwise be emitted into the environment. Examples of industrial flue gas include gases generated during the manufacture of ferrous and non-ferrous metal products, petroleum refining processes, coal gasification, biomass gasification, power generation, carbon black production, ammonia production, methanol production, and coke manufacturing.

[0061] Depending on the composition of the CO-containing substrate, the CO-containing substrate may be provided directly to the fermentation process or may be further modified to include a suitable H2:CO molar ratio. In one aspect, the H2:CO molar ratio of the CO-containing substrate provided to the fermenter is about 0.2 or more; in another aspect, about 0.25 or more; and in yet another aspect, about 0.5 or more. In another aspect, the CO-containing substrate provided to the fermenter may include about 40 mol% or more CO plus H2 and about 30 mol% or less CO; in another aspect, about 50 mol% or more CO plus H2 and about 35 mol% or less CO; and in yet another aspect, about 80 mol% or more CO plus H2 and about 20 mol% or less CO.

[0062] In one aspect, the CO-containing substrate primarily comprises CO and H2. In this aspect, the CO-containing substrate will contain at least about 10 mol% CO, in one aspect at least about 20 mol% CO, in another aspect about 10–about 100 mol% CO, in another aspect about 20–about 100 mol% CO, in another aspect about 30–about 90 mol% CO, in another aspect about 40–about 80 mol% CO, and in another aspect about 50–about 70 mol% CO. The CO / CO2 ratio of the CO-containing substrate is at least about 0.75, in another aspect at least about 1.0, and in another aspect at least about 1.5.

[0063] In one aspect, the gas separator is configured to substantially separate at least a portion of the gas stream, wherein this portion comprises one or more components. For example, the gas separator may separate CO2 from a gas stream comprising CO, CO2, and H2, wherein the CO2 may be passed to a CO2 remover, and the remaining gas stream (containing CO and H2) may be passed to a bioreactor. Any gas separator known in the art may be used. In one aspect, the syngas supplied to the fermenter will have about 10 mol% or less CO2, in another aspect about 1 mol% or less CO2, and in yet another aspect about 0.1 mol% or less CO2.

[0064] Some gas streams may include high concentrations of CO and low concentrations of H2. In one aspect, optimizing the composition of the substrate stream can be desirable to achieve more efficient alcohol production and / or overall carbon capture. For example, the concentration of H2 in the substrate stream can be increased before it is directed to the bioreactor.

[0065] According to a specific aspect of the invention, streams from two or more sources may be combined and / or blended to produce a desired and / or optimized substrate stream. For example, a stream containing a high concentration of CO (e.g., exhaust gas from a rolling mill converter) may be combined with a stream containing a high concentration of H2 (e.g., tail gas from a rolling mill coke oven).

[0066] Depending on the composition of the CO-containing gaseous substrate, it may be desirable to treat it before introducing it into fermentation to remove any undesirable impurities, such as dust particles. For example, the gaseous substrate can be filtered or washed using known methods.

[0067] Bioreactor design and operation The description of the fermenter design is based on U.S. Serial Nos. 13 / 471,827 and 13 / 471,858, filed May 15, 2012, and U.S. Serial No. 13 / 473,167, filed May 16, 2012, all of which are incorporated herein by reference.

[0068] According to one approach, the fermentation process is initiated by adding a culture medium to the reactor vessel. Examples of culture medium compositions are described in U.S. Serial Nos. 61 / 650,098 and 61 / 650,093, filed May 22, 2012, and U.S. Patent No. 7,285,402, filed July 23, 2001, all of which are incorporated herein by reference. The culture medium may be sterilized to remove unwanted microorganisms, and the reactor may be inoculated with the desired microorganisms. Sterilization may not always be necessary.

[0069] In one respect, the microorganisms used include acetic acid-producing bacteria. Examples of usable acetic acid-producing bacteria include Clostridium (…). Clostridium Acetic acid-producing bacteria, such as Clostridium youngii ( Clostridium ljungdahlii ) strains, including those described in WO 2000 / 68407, EP 117309, U.S. Patents 5,173,429, 5,593,886 and 6,368,819, WO 1998 / 00558 and WO2002 / 08438; Clostridium difficile (autoethanol-producing) Clostridium autoethanogenum (DSM 10061 and DSM 19630 of DSMZ, Germany) strains, including those described in WO 2007 / 117157 and WO 2009 / 151342; and Clostridium laurylidum ( Clostridium ragsdali (P11, ATCC BAA-622) and Bageophilus alkalophilus ( Alkalibaculum bacchi (CP11, ATCC BAA-1772), including those strains described in U.S. Patent 7,704,723 and "Biofuels and Bioproducts from Biomass-Generated Synthesis Gas," by Hasan Atiyeh, proposed at the Oklahoma EPSCoR Annual State Conference, April 29, 2010; and Clostridium carboxylate ( Clostridium carboxidivorans (ATCC PTA-7827), described in U.S. Patent Application 2007 / 0276447. Other applicable microorganisms include Mucor (…). Moorella ) genus, including species of Muller ( Moorella sp.) HUC22-1; and carboxylated thermophilic bacteria ( Carboxydothermus ) 。 These references are cited in this article. Mixed cultures of two or more microorganisms may be used.

[0070] Some examples of useful bacteria include Kavulinia acetogenes (Kevlaria acetogenes). Acetogenium kivui ), moist anaerobic acetic acid bacteria (Acetoanaerobium noterae Acetobacter wuerii (), Acetobacterium woodii ), Bacillus bacitratus ( Alkalibaculum bacchi CP11 (ATCC BAA-1772), Propionibacterium brevicornu ( Blautia producta ), methylbutyric acid bacteria ( Butyribacterium methylotrophicum ), underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous ), Pacific underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous pacificus ), hydrogen-producing carboxyl thermophilic bacteria ( Carboxydothermus hydrogenoformans Acetic acid bacteria () Clostridium aceticum Clostridium acetonebutanol ( Clostridium acetobutylicum Clostridium acetonebutanol ( Clostridium acetobutylicum P262 (DSM 19630 from DSMZ, Germany), self-produced Clostridium ethanoliferum ( Clostridium autoethanogenum (DSM 19630 of the German DSMZ) 、 Self-produced Clostridium ethanol ( Clostridium autoethanogenum (DSM 10061 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 23693 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 24138 from DSMZ, Germany), Clostridium carboxylate ( Clostridium carboxidivorans ) P7 (ATCC PTA-7827), Clostridium krill ( Clostridium coskatii (ATCC PTA-10522), Clostridium decoctioni ( Clostridium drakei Clostridium yangii ( Clostridium ljungdahlii ) PETC (ATCC 49587), Clostridium yangii ( Clostridium ljungdahlii ) ERI2 (ATCC 55380), Clostridium yangii ( Clostridium ljungdahlii ) C-01 (ATCC 55988), Clostridium yangii ( Clostridium ljungdahlii ) O-52 (ATCC 55889), Clostridium macrocarpa ( Clostridium magnum ), Clostridium pasteurellum ( Clostridium pasteurianum (DSM 525 from DSMZ in Germany), Clostridium laurylidum ( Clostridium ragsdali ) P11 (ATCC BAA-622), Clostridium paederi ( Clostridium scatologenes ), Clostridium thermophilum ( Clostridium thermoaceticum ), Clostridium Tunneloni ( Clostridiumultunense ), Kuhl's desulfurized enterobacteria ( Desulfotomaculum kuznetsovii ), Eubacterium mucilaginosa (Eubacterium limosum ), sulfur-reducing bacteria ( Geobacter sulfurreducens ), Methanophora acetophenones ( Methanosarcina acetivorans ), Pasteurella multocida ( Methanosarcina barkeri ), thermoacetic acid murine bacteria ( Morrella thermoacetica ), thermoautotrophic Murraya ( Morrella thermoautotrophica ), Fanny acetobacterium ( Oxobacter pfennigii ), producing Peptostreptococcus ( Peptostreptococcus productus ), producing rumen cocci ( Ruminococcus productus ), Kaiwu thermophilic anaerobic bacteria ( Thermoanaerobacter kivui ) and their mixtures.

[0071] Fermentation should ideally be carried out under suitable conditions that result in the desired fermentation (e.g., CO to ethanol). Reaction conditions to consider include pressure, temperature, gas flow rate, liquid flow rate, culture medium pH, culture medium redox potential, stirring rate (if using a stirred tank reactor), inoculum level, maximum gaseous substrate concentration to ensure that CO in the liquid phase does not become a limiting gas, and maximum product concentration to avoid product inhibition.

[0072] The method of the present invention can be used to maintain the viability of microbial cultures in which the microbial cultures are CO-limited, such that the rate of CO transfer into the solution is lower than the absorption rate of the culture. This can occur when a CO-containing substrate cannot be continuously supplied to the microbial culture; when the mass transfer rate is low; or when there is insufficient CO in the substrate stream to maintain culture viability at the optimized temperature. In such embodiments, the microbial culture will rapidly deplete the CO dissolved in the liquid nutrient medium and become substrate-limited because other substrates cannot be supplied sufficiently quickly.

[0073] start up Following inoculation, an initial feed gas supply rate is established to effectively supply the initial microbial community. Effluent gases are analyzed to determine their content. The results of the gas analysis are used to control the feed gas rate. In this respect, the method provides a calculated CO concentration:initial cell density ratio of approximately 0.5–approximately 0.9, in another respect, approximately 0.6–approximately 0.8, in another respect, approximately 0.5–approximately 0.7, and in another respect, approximately 0.5–approximately 0.6.

[0074] On the other hand, the fermentation method includes supplying syngas to the fermentation medium in an amount that effectively provides an initial calculated CO concentration of about 0.15 mM to about 0.70 mM in the fermentation medium, on the other hand, about 0.15 mM to about 0.50 mM, on the other hand, about 0.15 mM to about 0.35 mM, on the other hand, about 0.20 mM to about 0.30 mM, and on the other hand, about 0.23 mM to about 0.27 mM. The method effectively increases the cell density compared to the initial cell density.

[0075] The target cell density used in this article refers to a cell density of about 2.0 g / L or greater, in another respect, about 2 to about 30 g / L, in another respect, about 2 to about 25 g / L, in another respect, about 2 to about 20 g / L, in another respect, about 2 to about 10 g / L, in another respect, about 2 to about 8 g / L, in another respect, about 3 to about 30 g / L, in another respect, about 3 to about 6 g / L, in another respect, about 4 to about 5 g / L.

[0076] After startup Once the desired level is reached, the liquid phase and cell material are removed from the reactor and culture medium is replenished. This method effectively increases cell density to approximately 2.0 g / L or greater, on the other hand, approximately 2–approximately 30 g / L, on the other hand, approximately 2–approximately 25 g / L, on the other hand, approximately 2–approximately 20 g / L, on the other hand, approximately 2–approximately 10 g / L, on the other hand, approximately 2–approximately 8 g / L, on the other hand, approximately 3–approximately 30 g / L, on the other hand, approximately 3–approximately 6 g / L, and on the other hand, approximately 4–approximately 5 g / L.

[0077] Determining nutrient concentrations In one aspect, the method involves determining the concentration of nutrients in the fermentation medium. The monitored nutrient concentrations may include K, Mg, P, and mixtures thereof. In this regard, nutrient concentrations, specifically potassium concentrations, can be determined using ion chromatography (IC), inductively coupled plasma atomic absorption spectrometry (ICP), ion-selective electrodes, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof. Some examples of available methods include: Small, Hamish (1989). Ion chromatography (Ion Chromatography), New York: Plenum Press, ISBN 0-306-43290-0; Tatjana Weiss; Weiss, Joachim (2005). Handbook of Ion Chromatography(Handbook of Ion Chromatography), Weinheim: Wiley-VCH, ISBN 3-527-28701-9; Gjerde, Douglas T.; Fritz, James S, (2000) Ion Chromatography (Ion Chromatography), Weinheim:Wiley-VCH, ISBN 3-527-29914-9; Joachim Weiss, Tatjana Weiss (Translated by) (2005), Handbook of Ion Chromatography, Third, Completely Revised and Enlarged Edition, John Wiley and Sons, Inc, 931p, ISBN:3-527-28701-9; and Jackson, Peter; Haddad, Paul R, (1990). Ion chromatography:principles and applications (Ion Chromatography: Principles and Applications), Amsterdam: Elsevier, ISBN 0-444-88232-4, are incorporated herein by reference. Some examples of instruments that can be used include ICs available from Thermo Scientific Dionex (www.thermoscientific.com / dionex) and OFITE (Houston, TX - www.ofite.com).

[0078] In another aspect, the method may further include determining potassium, magnesium, and / or PO4 in the fermentation medium. -3 The concentration of potassium, magnesium, and PO4. In this regard, the method may include monitoring and controlling the concentrations of potassium, magnesium, and PO4. -3 Any one or any combination of elements. The table below provides an example of the elements and composition used.

[0079] Culture medium composition and feed rate control The composition of an effective culture medium is described in U.S. Patent Serial No. 13 / 889,700, filed May 8, 2013; and U.S. Patent Serial Nos. 13 / 890,324 and 13 / 890,777, both filed May 9, 2013, all of which are incorporated herein by reference.

[0080] The fermentation medium contains less than about 0.01 g / L of yeast extract and less than about 0.01 g / L of carbohydrates.

[0081] Sulfur is supplied to the fermentation process in the form of NaHS. In this regard, by adding an effective amount of NaHS to the fermentation medium, a H2S concentration of approximately 20 to approximately 500 ppm is maintained in the fermentation exhaust gas. In another aspect, approximately 200 to approximately 500 ppm H2S is maintained in the fermentation exhaust gas; in yet another aspect, approximately 250 to approximately 450 ppm H2S is maintained in the fermentation exhaust gas; and in yet another aspect, approximately 300 to approximately 400 ppm H2S is maintained in the fermentation exhaust gas.

[0082] The process operation maintains a pH range of approximately 3.5 to approximately 5.0, and on the other hand, approximately 4 to approximately 5. The nitrogen source (N) is provided via ammonium hydroxide, added as a separate feed stream under pH control; therefore, NH4+... + The dosage should be slightly exceeded by several hundred ppm.

[0083] According to one aspect of the method, first and second culture media are provided to fermentation. In this aspect, the first culture media is provided at a rate that effectively maintains a potassium concentration of about 20 to about 200 mg / L in the fermentation medium until a target cell density is reached. In another aspect, the potassium concentration in the fermentation medium is maintained at about 20 to about 160 mg / L; in another aspect, about 20 to about 100 mg / L; in another aspect, about 110 to about 190 mg / L; in another aspect, about 120 to about 180 mg / L; in another aspect, about 130 to about 170 mg / L; and in another aspect, about 140 to about 160 mg / L. When the target cell density is reached, the first and second culture media are provided at a rate that effectively maintains a potassium concentration of about 20 to about 160 mg / L in the fermentation medium; in another aspect, about 20 to about 50 mg / L; and in another aspect, about 30 to about 40 mg / L.

[0084] On the other hand, the culture medium is designed to effectively maintain approximately 10 to approximately 120 mg / L PO4. -3 The concentration rate is provided, in one aspect, about 10 to about 100 mg / L; in another aspect, about 10 to about 80 mg / L; in another aspect, about 10 to about 50 mg / L; and in another aspect, about 10 to about 25 mg / L. This is to effectively maintain about 10 to about 120 mg / L PO4 in the fermentation medium when the target cell density is reached. -3 The concentration rates provided by the first and second culture media are, in one aspect, about 10 to about 80 mg / L, in another aspect, about 10 to about 40 mg / L, and in yet another aspect, about 10 to about 25 mg / L.

[0085] In another aspect, the culture medium is provided at a rate that effectively maintains a Mg concentration of about 1 to about 6 mg / L, in another aspect, about 1 to about 4 mg / L, and in another aspect, about 1 to about 3 mg / L, until the target cell density is reached. Upon reaching the target cell density, the first and second culture media are provided at a rate that effectively maintains a magnesium concentration of about 1 to about 6 mg / L in the fermentation medium, in another aspect, about 1 to about 4 mg / L, and in another aspect, about 1 to about 3 mg / L.

[0086] In another respect, the culture medium is provided at a rate that effectively maintains a first culture medium to a second culture medium volume flow rate of about 15:1 to about 2:1, in another respect, about 10:1 to about 2:1, and in yet another respect, about 4:1 to about 2:1.

[0087] In another aspect, the method includes nutrient feed rate control to achieve the desired nutrient concentration. Specifically, if the potassium content is found to be greater than approximately 50 mg / L, the volume of the first culture medium is reduced by approximately 10% every 4 hours until the target potassium content is reached. In another aspect, if the phosphate content is found to be less than approximately 10 ppm, the volume of the first culture medium is increased by approximately 10% per hour until the target phosphate content is reached. In yet another aspect, if the magnesium content is found to be less than approximately 1 mg / L, the volume of the first culture medium is increased by approximately 10% per hour until the target magnesium content is reached.

[0088] The syngas fermentation method described herein, utilizing culture medium and acetic acid-producing bacteria in a bioreactor, is effective for converting CO in syngas into alcohols and other products. In this regard, ethanol production rate can be expressed as STY (space-time yield, expressed as g ethanol / (L·day)). In this regard, the method is effective for providing an STY (space-time yield) of at least about 10 g ethanol / (L·day). Possible STY values ​​include about 10 g ethanol / (L·day) to about 200 g ethanol / (L·day), in another aspect, about 10 g ethanol / (L·day) to about 160 g ethanol / (L·day), in another aspect, about 10 g ethanol / (L·day) to about 120 g ethanol / (L·day), in another aspect, about 10 g ethanol / (L·day) to about 80 g ethanol / (L·day), in another aspect, about 10 g ethanol / (L·day) to about 15 g ethanol / (L·day), in another aspect, about 15 g ethanol / (L·day) to about 20 g ethanol / (L·day). • day), in another aspect, about 20 g ethanol / (L·day) to about 140 g ethanol / (L·day), in another aspect, about 20 g ethanol / (L·day) to about 100 g ethanol / (L·day), in another aspect, about 40 g ethanol / (L·day) to about 140 g ethanol / (L·day), in another aspect, about 40 g ethanol / (L·day) to about 100 g ethanol / (L·day), in another aspect, about 10 g ethanol / (L·day), in another aspect, about 15 g ethanol / (L·day), in another aspect, about 16 g ethanol / (L·day).

[0089] Although the invention disclosed herein has been described through specific embodiments, examples and applications, those skilled in the art can make many modifications and variations thereto without departing from the scope of the invention as set forth in the claims.

Claims

1. A method for controlling the fermentation of a CO-containing substrate, the method comprising: Provide CO-containing substrates to fermentation and enable the CO-containing substrates to ferment; and Nutrient concentrations in fermentation media can be monitored using techniques selected from the following: ion chromatography, inductively coupled plasma atomic absorption spectrometry, ion-selective electrode, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof. The method described herein provides a target cell density of approximately 3 to approximately 30 g / L. The fermentation medium comprises first and second nutrient-containing media. The first media is provided to fermentation at a rate that effectively maintains a potassium concentration of approximately 120 to approximately 180 mg / L until the target cell density is reached. After reaching the target cell density, the first media is provided to fermentation at a rate that effectively maintains a potassium concentration of approximately 20 to approximately 50 mg / L. The first media is also provided to fermentation at a rate that effectively maintains a magnesium concentration of approximately 1 to approximately 6 mg / L until the target cell density is reached. After reaching the target cell density, the first media is provided to fermentation at a rate that effectively maintains a magnesium concentration of approximately 1 to approximately 4 mg / L. The second media is provided to fermentation at a rate that effectively maintains a first media to second media volume flow rate ratio of approximately 15:1 to approximately 2:

1. The monitored nutrients were selected from K, Mg, and PO4. -3 Fe, Zn, Ni, Co and mixtures thereof, wherein the first culture medium comprises K, Mg, Fe, PO4 -3 The second culture medium contains elements selected from W, Se, and mixtures thereof, and the second culture medium contains elements selected from W, Se, and mixtures thereof.

2. The method of claim 1, wherein the monitored nutrient is K or PO4. -3 Mg and its mixtures.

3. The method of claim 1, wherein the technique used to monitor nutrient concentration is ion chromatography.

4. The method of claim 1, wherein the first culture medium is used to effectively maintain a concentration of about 10 to about 120 mg / L PO4 in the culture medium. -3 The concentration is supplied at a rate that continues to ferment until the target cell density is reached.

5. The method of claim 4, wherein after the target cell density is reached, the first culture medium is used to effectively maintain a concentration of about 10 to about 100 mg / L PO4. -3 The concentration rate is provided to the fermentation process.

6. The method of claim 1, wherein the CO-containing substrate has a CO / CO2 molar ratio of at least about 0.

75.

7. The method of claim 1, wherein the CO-containing substrate has about 20 to about 100% molar CO.

8. The method of claim 1, wherein the pH of the fermentation medium is maintained at a level of about 3.5 to about 5.

0.

9. The method of claim 8, wherein the pH is maintained by adding ammonium hydroxide.

10. The method of claim 1, further comprising maintaining an H2S concentration in the fermentation exhaust gas in the range of about 20 to about 500 ppm H2S.

11. The method of claim 10, wherein by adding NaHS to the fermentation medium, the H2S concentration in the fermentation exhaust gas is maintained at about 20 to about 500 ppm.

12. The method of claim 1, wherein the fermentation comprises acetic acid-producing bacteria.

13. The method of claim 12, wherein the acetic acid-producing bacteria is selected from Kavulacillus acetogenes (Kaiwu acetogenes). Acetogenium kivui ), moist anaerobic acetic acid bacteria ( Acetoanaerobium noterae Acetobacter wuerii (), Acetobacterium woodii ), Bacillus bacitratus ( Alkalibaculum bacchi CP11 (ATCC BAA-1772), Propionibacterium brevicornu ( Blautia producta ), methylbutyric acid bacteria ( Butyribacterium methylotrophicum ), underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous ), Pacific underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous pacificus ), hydrogen-producing carboxyl thermophilic bacteria ( Carboxydothermus hydrogenoformans Acetic acid bacteria () Clostridium aceticum Clostridium acetonebutanol ( Clostridium acetobutylicum Clostridium acetonebutanol ( Clostridium acetobutylicum P262 (DSM 19630 from DSMZ, Germany), self-produced Clostridium ethanoliferum ( Clostridium autoethanogenum (DSM 19630 of the German DSMZ) 、 Self-produced Clostridium ethanol ( Clostridium autoethanogenum (DSM 10061 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 23693 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 24138 from DSMZ, Germany), Clostridium carboxylate ( Clostridium carboxidivorans ) P7 (ATCC PTA-7827), Clostridium krill ( Clostridium coskatii (ATCC PTA-10522), Clostridium decoctioni ( Clostridium drakei Clostridium yangii ( Clostridium ljungdahlii ) PETC (ATCC 49587), Clostridium yangii ( Clostridium ljungdahlii ) ERI2 (ATCC 55380), Clostridium yangii ( Clostridium ljungdahlii ) C-01 (ATCC 55988), Clostridium yangii ( Clostridium ljungdahlii ) O-52 (ATCC55889), Clostridium macrocarpa ( Clostridium magnum ), Clostridium pasteurellum ( Clostridium pasteurianum (DSM 525 from DSMZ, Germany), Clostridium laurylidum ( Clostridium ragsdali ) P11 (ATCC BAA-622), Clostridium paederi ( Clostridium scatologenes ), Clostridium thermophilum ( Clostridium thermoaceticum ), Clostridium Tunneloni ( Clostridium ultunense ), Kuhl's desulfurized enterobacteria ( Desulfotomaculum kuznetsovii ), Eubacterium mucilaginosa ( Eubacterium limosum ), sulfur-reducing bacteria ( Geobacter sulfurreducens ), Methanophora acetophenones ( Methanosarcina acetivorans ), Pasteurella multocida ( Methanosarcina barkeri ), thermoacetic acid murine bacteria ( Morrella thermoacetica ), thermoautotrophic Murraya ( Morrella thermoautotrophica ), Fanny acetobacterium ( Oxobacter pfennigii ), producing Peptostreptococcus ( Peptostreptococcus productus ), producing rumen cocci ( Ruminococcus productus ), Kaiwu thermophilic anaerobic bacteria ( Thermoanaerobacter kivui ) and their mixtures.

14. A method for fermenting a CO-containing substrate, the method comprising: Provide CO-containing substrates to fermentation and enable the CO-containing substrates to ferment; Determine the potassium concentration in the fermentation medium; and A first culture medium and a second culture medium are provided to the fermentation process. The first culture medium is provided at a rate that effectively maintains a potassium concentration in the range of about 120 to about 180 mg / L until a target cell density is reached. After the target cell density is reached, the first culture medium is provided to the fermentation process at a rate that effectively maintains a potassium concentration in the culture medium of about 20 to about 50 mg / L. The first culture medium is also provided to the fermentation process at a rate that effectively maintains a magnesium concentration in the culture medium of about 1 to about 6 mg / L until the target cell density is reached. After the target cell density is reached, the first culture medium is provided to the fermentation process at a rate that effectively maintains a magnesium concentration in the culture medium of about 1 to about 4 mg / L. The second culture medium is provided to the fermentation process at a rate that effectively maintains a first culture medium to second culture medium volumetric flow rate ratio of about 15:1 to about 2:

1. The target cell density is approximately 3 to approximately 30 g / L. The first culture medium includes substances selected from K, Mg, Fe, and PO4. -3 The second culture medium contains elements selected from W, Se, and mixtures thereof, and the second culture medium contains elements selected from W, Se, and mixtures thereof.

15. The method of claim 14, wherein the CO-containing substrate has a CO / CO2 molar ratio of at least about 0.

75.

16. The method of claim 14, wherein the CO-containing substrate has about 20 to about 100% molar CO.

17. The method of claim 14, wherein the concentration of potassium in the culture medium is determined by a method selected from: ion chromatography, inductively coupled plasma atomic absorption spectrometry, ion-selective electrode, flame photometry, flame ionization atomic absorption spectrometry, and combinations thereof.

18. The method of claim 14, wherein the pH of the fermentation medium is maintained at a level of about 3.5 to about 5.

0.

19. The method of claim 14, wherein the pH is maintained by adding ammonium hydroxide.

20. The method of claim 14, further comprising maintaining an H2S concentration in the fermentation exhaust gas in the range of about 20 to about 500 ppm H2S.

21. The method of claim 20, wherein by adding NaHS to the fermentation medium, the H2S concentration in the fermentation exhaust gas is maintained at about 20 to about 500 ppm.

22. The method of claim 14, wherein the fermentation comprises acetic acid-producing bacteria.

23. The method of claim 22, wherein the acetic acid-producing bacteria is selected from Kavulacillus acetogenes (Kaiwu acetogenes). Acetogenium kivui ), moist anaerobic acetic acid bacteria ( Acetoanaerobium noterae Acetobacter wuerii (), Acetobacterium woodii ), Bacillus bacitratus ( Alkalibaculum bacchi CP11 (ATCC BAA-1772), Propionibacterium brevicornu ( Blautia producta ), methylbutyric acid bacteria ( Butyribacterium methylotrophicum ), underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous ), Pacific underground thermophilic anaerobic bacteria ( Caldanaerobacter subterraneous pacificus ), hydrogen-producing carboxyl thermophilic bacteria ( Carboxydothermus hydrogenoformans Acetic acid bacteria () Clostridium aceticum Clostridium acetonebutanol ( Clostridium acetobutylicum Clostridium acetonebutanol ( Clostridium acetobutylicum P262 (DSM 19630 from DSMZ, Germany), self-produced Clostridium ethanoliferum ( Clostridium autoethanogenum (DSM 19630 of the German DSMZ) 、 Self-produced Clostridium ethanol ( Clostridium autoethanogenum (DSM 10061 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 23693 from DSMZ, Germany), self-produced Clostridium ethanolae ( Clostridium autoethanogenum (DSM 24138 from DSMZ, Germany), Clostridium carboxylate ( Clostridium carboxidivorans ) P7 (ATCC PTA-7827), Clostridium krill ( Clostridium coskatii (ATCC PTA-10522), Clostridium decoctioni ( Clostridium drakei Clostridium yangii ( Clostridium ljungdahlii ) PETC (ATCC 49587), Clostridium yangii ( Clostridium ljungdahlii ) ERI2 (ATCC 55380), Clostridium yangii ( Clostridium ljungdahlii ) C-01 (ATCC 55988), Clostridium yangii ( Clostridium ljungdahlii ) O-52 (ATCC55889), Clostridium macrocarpa ( Clostridium magnum ), Clostridium pasteurellum ( Clostridium pasteurianum (DSM 525 from DSMZ, Germany), Clostridium laurylidum ( Clostridium ragsdali ) P11 (ATCC BAA-622), Clostridium paederi ( Clostridium scatologenes ), Clostridium thermophilum ( Clostridium thermoaceticum ), Clostridium Tunneloni ( Clostridium ultunense ), Kuhl's desulfurized enterobacteria ( Desulfotomaculum kuznetsovii ), Eubacterium mucilaginosa ( Eubacterium limosum ), sulfur-reducing bacteria ( Geobacter sulfurreducens ), Methanophora acetophenones ( Methanosarcina acetivorans ), Pasteurella multocida ( Methanosarcina barkeri ), thermoacetic acid murine bacteria ( [[ID=(此处疑似序号有误,应为27)]]Morrella thermoacetica ), thermoautotrophic Murraya ( Morrella thermoautotrophica ), Fanny acetobacterium ( Oxobacter pfennigii ), producing Peptostreptococcus ( Peptostreptococcus productus ), producing rumen cocci ( Ruminococcus productus ), Kaiwu thermophilic anaerobic bacteria ( Thermoanaerobacter kivui ) and their mixtures.

24. The method of claim 14, wherein the method is effective to provide a STY of 10 g total alcohol / (L·day) or greater.

25. The method of claim 14, further comprising maintaining a concentration of about 10 to about 120 mg / L PO4 in the culture medium effectively. -3 The concentration rate provides the first culture medium to the fermentation process until the target cell density is reached.

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