Use of oxyhydrogen microorganisms for the recovery and conversion of non-photosynthetic carbon from inorganic carbon sources and / or C1 carbon sources into useful organic compounds.

Abstract

The present invention provides compositions and methods for a hybrid biochemical process that recovers and converts organic chemicals (biofuels or other valuable biomass, chemicals, industrial products, or pharmaceuticals) from a mixture containing carbon dioxide and / or other forms of inorganic carbon and / or a C1 carbon source (e.g., carbon monoxide, methane, methanol, formate, or formic acid) and / or C1 chemicals (e.g., various syngas compositions, but not limited thereto). [Solution] In certain embodiments, the present invention utilizes microorganisms capable of performing oxyhydrogen reactions and autotrophic CO2 fixation in one or more steps of the process to fix an inorganic carbon source or a C1 carbon source into an organic chemical with a longer carbon chain.

Description

[Technical Field]

[0001] Related applications This application claims priority under U.S. Provisional Patent Application No. 61 / 328,184, filed April 27, 2010, entitled “Use of Oxyhydrogen Microorganisms for Recovery and Conversion of Non-Photosynthetic Carbon from Inorganic Carbon Sources to Useful Organic Compounds,” pursuant to Section 119(e) of the U.S. Patent Act. This application is also a continuation-in-part application of International Patent Application PCT / US2010 / 001402, filed on 12 May 2010, entitled “Biochemical Process Using Chemoautotrophic Microorganisms for Chemosynthesis and Immobilization of Organic Compounds from Carbon Dioxide and / or Other Inorganic Carbon Sources and Production of Other Useful Products,” which is a continuation-in-part application of U.S. Patent Application No. 12 / 613,550, filed on 6 November 2009, entitled “Biochemical Process Using Chemoautotrophic Microorganisms for Chemosynthesis and Immobilization of Organic Compounds from Carbon Dioxide and / or Other Inorganic Carbon Sources and Production of Other Useful Products,” which claims the benefits of U.S. Provisional Patent Application No. 61 / 111,794, filed on 6 November 2008, entitled “Biochemical Process Using Chemoautotrophic Microorganisms for Carbon Cycling by Chemosynthesis of Biofuels and Other Useful Products from Carbon Dioxide and Other Inorganic Carbon Sources.” Each of these applications is incorporated herein by reference in its entirety for all purposes. [Background technology]

[0002] The present invention belongs to the technical fields of biofuels, bioremediation, carbon capture, carbon dioxide fuel conversion, carbon cycle, carbon sequestration, energy storage, gas liquefaction, waste-to-energy fuel conversion, syngas conversion, and renewable / alternative energy and / or low-carbon emission energy. Specifically, the present invention is a unique use of biocatalysts in biochemical processes that fix carbon dioxide and / or other forms of inorganic carbon sources and / or other C1 carbon sources into longer-chain organic chemicals in non-photosynthetic processes powered by low-carbon emission energy sources and / or waste energy sources. In addition, the present invention encompasses the production of chemical by-products that are co-feeded by carbon fixation reaction steps and / or non-biological reaction steps as part of a total carbon capture and conversion process or syngas conversion process. The present invention can enable the efficient and economical capture of carbon dioxide from the atmosphere or from point emissions of carbon dioxide, as well as the economical use of waste energy sources and / or renewable energy sources and / or low-carbon emission energy sources, for the production of transport liquid fuels and / or other organic chemicals. Therefore, it will help address climate change caused by greenhouse gases, as well as contribute to the domestic production of renewable liquid fuels and / or other organic chemicals for transport that do not depend on agriculture.

[0003] Significant interest and effort have been devoted to developing technologies that use renewable or waste-to-energy to convert carbon dioxide or other low-value carbon sources into useful organic chemicals, in order to provide alternatives to chemicals, materials, and fuels derived from petroleum or other fossil sources. In the field of CO2 conversion, the focus has been almost entirely on biological methods utilizing photosynthesis to fix CO2 into biomass or end products, while some effort has also been devoted to entirely abiotic chemical processes for fixing CO2.

[0004] A relatively under-received type of CO2 organic chemicalization method is the hybrid chemical / biological process. In this case, the biological process is limited to CO2 fixation, which corresponds to the dark reaction of photosynthesis. Potential advantages of such hybrid CO2 organic chemicalization processes include the ability to power the process by combining CO2 fixation enzyme functions acquired through billions of years of evolution with a wide range of abiotic technologies such as solar photovoltaic (PV), solar thermal, wind, geothermal, hydroelectric, or nuclear power. Microorganisms that fix carbon without the use of light can be confined to a more controlled and protected environment, less prone to water and nutrient loss, pollution, and weather-related disasters than those usable for culturing photosynthetic microorganisms. Furthermore, increased bioreactor capacity can be adapted to vertical rather than horizontal configurations, potentially leading to even greater land-use efficiency. Hybrid chemical / biological systems offer the potential for CO2 organic chemicalization processes that retain the biological functions of complex organic synthesis from CO2 while avoiding many of the drawbacks of photosynthesis.

[0005] Chemoautotrophic microorganisms are generally microorganisms capable of CO2 fixation in a similar manner to the dark-light reaction of photosynthesis, but these microorganisms can obtain the reducing equivalents necessary for CO2 fixation from an external source and do not need to produce them internally via the light-light reaction of photosynthesis. The carbon fixation biochemical pathways carried out by chemoautotrophic organisms include the reductive tricarboxylic acid cycle, the Calvin-Benson-Bassham cycle, and the Wood-Ljungdahl pathway. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Prior studies are known regarding the specific application of chemoautotrophic microorganisms in the recovery and conversion of CO2 gas to fixed carbon. However, many of these studies have suffered from limitations in the effectiveness, economic feasibility, practicality, and commercialization of the described processes. In certain embodiments, the present invention addresses one or more of the above-mentioned drawbacks. [Means for solving the problem]

[0007] The present invention, which utilizes oxyhydrogen microorganisms for the chemosynthesis and fixation of CO2 under carefully controlled oxygen levels, is considered to have advantages in producing long-chain organic compounds (e.g., C5 or higher). Since the energy density (energy per unit volume) is generally high in long-chain organic compounds, and their compatibility with current transport systems is generally greater compared to short-chain products such as C1 and C2 products, the ability to produce long-chain organic compounds is a significant advantage of the present invention.

[0008] In addressing the needs in the art that the inventors have identified in carrying out this invention, we describe a novel combined biochemical process for the recovery and conversion of inorganic carbon sources and / or C1 carbon sources into long-chain organic compounds, specifically organic compounds having a chain length of C5 or longer, via the use of oxyhydrogen microorganisms for carbon capture and fixation. In some embodiments, the process can be combined with the efficient production of high-value organic compounds, such as liquid hydrocarbon fuels, with the disposal of waste carbon sources and, furthermore, CO2 capture, which can provide an additional revenue source.

[0009] In one embodiment, a biochemical method for the recovery and conversion of inorganic carbon compounds and / or organic compounds containing only one carbon atom to organic chemicals is described. In some embodiments, the method includes introducing inorganic carbon compounds and / or organic compounds containing only one carbon atom into an environment suitable for the retention of oxyhydrogen microorganisms and / or capable of retaining extracts of oxyhydrogen microorganisms, and converting the inorganic carbon compounds and / or organic compounds containing only one carbon atom into organic chemicals and / or precursors thereof via at least one chemosynthetic carbon fixation reaction using oxyhydrogen microorganisms and / or cell extracts containing enzymes derived from oxyhydrogen microorganisms within that environment. In some embodiments, the chemosynthetic fixation reaction is driven at least in part by chemical and / or electrochemical energy provided by electron donors and electron acceptors that are chemically and / or electrochemically generated and / or introduced into the environment from at least one external source outside the environment.

[0010] In one embodiment, a bioreactor is described. In one group of embodiments, the bioreactor includes a first column comprising an upper and lower portion, and a second column comprising an upper and lower portion, wherein the upper portion of the second column is fluidly connected to the upper portion of the first column, and the lower portion of the second column is fluidly connected to the lower portion of the first column. In some embodiments, the bioreactor is constructed and arranged such that the volume of gas at the top of the first and / or second column is substantially steady when the liquid is circulated between the first and second columns. In some embodiments, the volume of gas occupies at least about 2% of the total volume of the column in which its volume is located.

[0011] In other embodiments, a method for operating a bioreactor is provided. In some embodiments, the method includes circulating a liquid containing a growth medium between a first column and a second column, wherein during operation, the volume of gas is maintained substantially steadily at the top of the first and / or second column, and the volume of gas occupies at least about 2% of the total volume of the column in which its volume is located.

[0012] In one embodiment, an electrolytic apparatus is provided. In some embodiments, the electrolytic apparatus includes a chamber constructed and arranged to electrolyze water to produce oxygen and hydrogen, and an outlet section including a separator constructed and arranged to separate at least a portion of the oxygen in the stream from at least a portion of the hydrogen in the stream so that the hydrogen content of the fluid exiting the separator is suitable for use as a feed stream to a reactor containing a culture of oxyhydrogen microorganisms.

[0013] In other embodiments, a method for operating an electrolytic apparatus is described. In some embodiments, the method includes electrolyzing water to produce a first stream containing oxygen and hydrogen, and separating at least a portion of the oxygen from at least a portion of the hydrogen to produce a second stream that is relatively richer in hydrogen than the first stream (the second stream being suitable as a supply stream to a reactor containing a culture of oxyhydrogen microorganisms).

[0014] In certain embodiments, the present invention provides compositions and methods for recovering carbon dioxide from carbon dioxide-containing gas streams and / or atmospheric carbon dioxide or dissolved, liquefied, or chemically bonded carbon dioxide via a biochemical process utilizing cell extracts containing biased or facultative oxyhydrogen microorganisms and / or enzymes derived from oxyhydrogen microorganisms in one or more carbon fixation process steps.

[0015] In certain embodiments, the present invention provides compositions and methods for converting a C1 chemical into a long-chain organic compound using a C1 carbon source (for example, carbon monoxide, methane, methanol, formate, or formic acid, but not limited thereto) and / or a mixture containing a C1 chemical (for example, various syngas compositions produced from various fixed carbon source materials that have been gasified, pyrolyzed, or steam reformed, but not limited thereto).

[0016] In certain embodiments, the present invention provides compositions and methods for the recovery, treatment, and use of organic compounds produced by immobilizing inorganic carbon sources and / or C1 carbon sources into long-chain organic compounds through chemosynthesis reactions carried out by oxyhydrogen microorganisms. In certain embodiments, the present invention provides compositions and methods for maintaining and controlling oxygen levels in a carbon fixation environment in order to produce improved (e.g., optimal) C5 or higher organic compound products via carbon fixation. In certain embodiments, the present invention provides compositions and methods for the generation, treatment, and delivery of chemical nutrients required for carbon fixation and maintenance of oxyhydrogen microbial cultures (e.g., providing electron donors and electron acceptors required for non-photosynthetic carbon fixation). In certain embodiments, the present invention provides compositions and methods for maintaining an environment conducive to carbon fixation and for the recovery and recycling of unused chemical nutrients and process water.

[0017] In certain embodiments, the present invention relates to a chemical process step carried out in series and / or parallel with a chemical synthesis reaction step, namely, a step of converting unrefined crude input chemicals into more refined chemicals suitable for supporting a chemical synthesis carbon fixation step; a step of converting energy input into a chemical form usable to drive chemical synthesis, specifically chemical energy in the form of electron donors and electron acceptors; directing inorganic carbon recovered from industrial, atmospheric, or aquatic sources to the carbon fixation step of the process under conditions suitable for supporting chemical synthesis carbon fixation by oxyhydrogen microorganisms or enzymes, and / or carbon monoxide, usable by oxyhydrogen microorganisms as a carbon source and optional energy source for synthesizing long-chain organic chemicals. The present invention provides compositions and methods for a process that directs C1 chemicals and / or mixtures containing C1 chemicals derived from low-value or waste carbon sources such as tahn, methanol, formate, or formic acid (for example, various syngas compositions derived from the gasification, pyrolysis, or steam reforming of various low-value or waste carbon sources, etc.), and processes that further process the output products of a carbon fixation process to form a form suitable for storage, transport, and sale, and / or safe disposal, resulting in a net reduction of gaseous CO2 released into the atmosphere and / or an upgrade from low-value or waste materials to finished chemicals, fuels, or nutrients. A fully chemical process process combined with a chemically synthesized carbon fixation process constitutes a total carbon recovery and conversion process according to some embodiments of the present invention.

[0018] One feature of certain embodiments of the present invention is the incorporation of one or more process steps into a chemical process for the recovery of inorganic carbon and conversion to fixed carbon products, utilizing oxyhydrogen microorganisms and / or enzymes derived from oxyhydrogen microorganisms as biocatalysts for fixing carbon dioxide and / or dissolved or solid inorganic carbon in a carbon dioxide-containing gas stream, or in the atmosphere or water, into organic compounds. In some such embodiments, dissolved carbon dioxide, inorganic carbon as carbonate ions or bicarbonate ions in a solution including a carbon dioxide-containing flue gas or process gas, or air or an aqueous solution such as seawater, or inorganic carbon in a solid phase (e.g., carbonates and bicarbonates) is added to a tank or sealed container containing a nutrient medium and oxyhydrogen microorganisms by pump injection or other means. In some such cases, oxyhydrogen microorganisms use one or more of the following electron donors, provided by pumping into a nutrient medium or otherwise, the chemical energy stored in molecular hydrogen and / or valence or conduction electrons in a solid electrode material and / or the following: ammonia, ammonium, carbon monoxide, dithionite, elemental sulfur, hydrocarbons, metabisulfite, nitrogen oxides, nitrites, sulfates such as thiosulfates (e.g., sodium thiosulfate (Na2S2O3) or calcium thiosulfate (CaS2O3)), sulfides such as hydrogen sulfide, sulfites, thionites, thionic acid, transition metals or their sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, phosphates, sulfates, or carbonates (but not limited to these) in the soluble or solid phase to perform chemosynthesis to fix inorganic carbon and convert it into an organic compound. In some embodiments, it is possible to use electrons in the conduction band or valence band in the solid electrode material. Electron donors can be oxidized by electron acceptors in chemical synthesis reactions.Examples of electron acceptors that can be used in chemical synthesis reaction steps include oxygen and / or other electron acceptors (for example, one or more of the following, namely carbon dioxide, ferric ions or other transition metal ions, nitrates, nitrites, sulfates, oxygen, or holes in the valence band or conduction band of solid electrode materials, but not limited to these).

[0019] One feature of certain embodiments of the present invention is the incorporation of one or more process steps into a chemical process for the conversion of a C1 carbon source (e.g., carbon monoxide, methane, methanol, formate, or formic acid) and / or a mixture containing a C1 chemical (e.g., various syngas compositions produced from various fixed carbon feedstocks that have been gasified, pyrolyzed, or steam-reformed), utilizing oxyhydrogen microorganisms and / or enzymes derived from oxyhydrogen microorganisms as a biocatalyst for the conversion of a C1 chemical to a long-chain organic chemical (i.e., carbon chain molecules of C2 or higher, and in some embodiments, carbon chain molecules of C5 or higher). In some such embodiments, the C1-containing syngas, or process gas, or a C1 chemical in pure liquid form or dissolved in solution, is added to a tank or sealed container containing a nutrient medium and oxyhydrogen microorganisms by pump injection or other means. In some such cases, oxyhydrogen microorganisms use the chemical energy stored in the C1 chemical and / or molecular hydrogen, as well as valence or conduction electrons in the solid electrode material, and / or electron donors provided by pumping or otherwise into the nutrient medium, namely, sulfates such as ammonia, ammonium, carbon monoxide, dithionite, elemental sulfur, hydrocarbons, metabisulfite, nitrous oxide, nitrite, thiosulfates (e.g., sodium thiosulfate (Na2S2O3) or calcium thiosulfate (CaS2O3)), sulfides such as hydrogen sulfide, sulfites, thionites, thionic acid, transition metals or their sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, sulfates, or carbonates (but not limited to these) in the soluble or solid phase to biochemically synthesize and extend the C1 chemical into an organic chemical with a longer carbon chain. Electron donors can be oxidized by electron acceptors in chemical synthesis reactions.Examples of electron acceptors that can be used in this reaction step include oxygen and / or other electron acceptors (e.g., one or more of the following: carbon dioxide, ferric ions or other transition metal ions, nitrates, nitrites, oxygen, or holes in the solid electrode material, but not limited thereto).

[0020] One or more chemical synthesis reaction steps of a process in which carbon dioxide and / or inorganic carbon is fixed into organic carbon in the form of organic compounds and biomass, and / or C1 chemicals (e.g., carbon monoxide, methane, methanol, formate, or formic acid, but not limited thereto) and / or mixtures containing C1 chemicals (e.g., various syngas compositions produced from various fixed carbon feedstocks that have been gasified, pyrolyzed, or steam reformed, but not limited thereto) are biochemically converted into long-chain organic chemicals (i.e., carbon chain molecules with C2 or more, and in some embodiments C5 or more), the reaction step of converting C1 chemicals into long-chain organic chemicals can be carried out under aerobic, microaerobic, anoxic, anaerobic conditions or facultative conditions. A facultative environment is considered to have an aerobic upper layer and an anaerobic lower layer due to the stratification of the water column.

[0021] In some embodiments of the present invention, the oxygen level is controlled such that the production of the target organic compound by hydrogenotrophic microorganisms via carbon fixation is controlled (e.g., optimized). One purpose of controlling the oxygen level is to control (e.g., optimize) the intracellular adenosine triphosphate (ATP) concentration via the reduction of oxygen by the cells and the production of ATP by oxidative phosphorylation, and at the same time to keep the environment sufficiently reducing so as to maintain a high ratio of NADH (or NADPH) to NAD (or NADP).

[0022] The advantage of using hydrogenotrophic microorganisms rather than strictly anaerobic acetate-producing or methane-producing microorganisms for carbon recovery applications and / or syngas conversion applications is that hydrogenotrophic microorganisms have higher oxygen tolerance.

[0023] An additional advantage of using hydrogenotrophic microorganisms rather than acetogenic bacteria for carbon recovery applications and / or syngas conversion applications and / or biofuel production is that, due to the production of ATP powered by the hydrogen reaction, instead of acetate products and butyrate products of acid production that can harm microorganisms due to a decrease in solution pH or accumulation to toxic levels, which are generally undesirable, it produces water products that can be easily incorporated into the process stream.

[0024] Further features of certain embodiments of the present invention relate to the source, generation, or recycling of electron donors used by hydrogenotrophic microorganisms that fix carbon dioxide into organic compounds and / or synthesize organic molecules with longer carbon chains from C1 chemicals. The electron donors used for carbon dioxide recovery and carbon fixation, in certain embodiments of the present invention, can be electrochemically or thermochemically generated or recycled using power based on a variety of renewable energy technologies and / or low carbon emission energy technologies, including but not limited to, photovoltaic power, solar thermal, wind power, hydroelectric power, nuclear power, geothermal, enhanced geothermal, ocean thermal, wave power, tidal power. The electron donors can also be of mineral origin, including but not limited to, minerals containing reduced S and Fe. The electron donors used in certain embodiments of the present invention can also be generated or recycled through a chemical reaction with hydrocarbons, which may or may not be non-renewable fossil fuels, provided that the chemical reaction results in little or no carbon dioxide gas emissions. For example, the redox reactions that produce carbonate products and hydrogen products that can be used as electron donors in the carbon fixation reaction step according to certain embodiments of the present invention are 2CH4+Fe2O3+3H2O→2FeCO3+7H2 and / or CH4+CaO+2H2O→CaCO3+4H2 include.

[0025] A further feature of certain embodiments of the present invention relates to the formation and recovery of organic compounds and / or biomass from one or more chemically synthesized carbon fixation steps. These organic compounds and / or biomass products may have a variety of applications.

[0026] A further feature of certain embodiments of the present invention relates to the use of oxyhydrogen microorganisms modified to produce organic compounds, biochemicals, or biomass in superior quantities and / or quality through chemosynthesis in one or more carbon fixation processes. The oxyhydrogen microorganisms used in these processes can be modified through artificial means, including but not limited to accelerated mutagenesis (e.g., using ultraviolet light treatment or chemical treatment), genetic engineering or genetic modification, hybridization, synthetic biology, or traditional selective breeding. Possible modifications of oxyhydrogen microorganisms include, but are not limited to, those aimed at producing organic compounds and / or biomass in increased quantities and / or quality for use as biofuels or feedstocks for producing biofuels (e.g., JP-8 jet fuel, diesel oil, gasoline, biodiesel oil, butanol, ethanol, hydrocarbons, methane, and pseudo-vegetable oils, or any other hydrocarbons suitable for use as renewable / alternative fuels that result in reduced greenhouse gas emissions).

[0027] Furthermore, compositions and methods for reducing the risk of gas fermentation using a mixture of hydrogen and oxygen within the process according to the present invention are also described.

[0028] Furthermore, compositions and methods that take advantage of the oxygen tolerance and ability of oxyhydrogen microorganisms to use oxygen as an electron acceptor are described to enable a system for converting water into a hydrogen electron donor or hydride electron donor and oxygen electron acceptor, which has improved efficiency compared to applying current state-of-the-art electrolysis techniques for the purpose of generating hydrogen electron donors or hydride electron donors and oxygen electron acceptors.

[0029] Furthermore, the process steps for recovering useful chemicals generated in both the biological carbon fixation and non-biological process steps, as well as for further finishing treatments, are also described.

[0030] Other advantages and novel features of the present invention will become apparent from the following detailed description of various embodiments (but not limited to) of the invention, when considered in conjunction with the accompanying drawings. All publications, patent applications, and patents cited herein are incorporated by reference in their entirety. In the event that this specification and the documents incorporated by reference contain conflicting and / or contradictory disclosures, this specification shall prevail.

[0031] Embodiments of the present invention will be described by reference to the accompanying diagrams, which are schematic and not intended to be drawn to actual size, but are not limited thereto. For clarity, not all elements are shown in all diagrams, nor are all elements of each embodiment of the present invention shown where not an example is necessary for a person skilled in the art to understand the invention. [Brief explanation of the drawing]

[0032] [Figure 1] This is a general process flow diagram of one embodiment of a carbon capture and sequestration process. [Figure 2] This is a process flow diagram of another embodiment of the present invention, which involves CO2 recovery carried out by microorganisms (e.g., hydrogen-oxidizing purple non-sulfur bacteria) capable of performing oxyhydrogen reactions to produce lipid-rich biomass that can be converted into JP-8 jet fuel. [Figure 3] This is a design for a bioreactor that utilizes the low solubility of hydrogen and oxygen gases in water, and avoids the dangerous mixing of hydrogen and oxygen by providing oxygen and hydrogen, which are necessary for cellular energy and carbon fixation, to oxyhydrogen microorganisms. [Figure 4]This is a design for a bioreactor that takes advantage of the relatively high solubility of carbon dioxide and the high ability of oxyhydrogen microorganisms to recover carbon dioxide from relatively dilute streams using a carbon concentration mechanism (CCM) in order to remove CO2 from a dilute gas mixture and separate it from low-solubility gases such as oxygen and nitrogen. [Figure 5] This electrolysis technology is specifically designed to leverage the advantages of oxyhydrogen microorganisms' tolerance and need for specific oxygen concentrations by reducing the complete separation of hydrogen and oxygen produced from standard electrolysis. [Modes for carrying out the invention]

[0033] The present invention provides compositions and methods for recovering and fixing carbon dioxide from carbon dioxide-containing gas streams and / or atmospheric carbon dioxide or liquefied or chemically bonded carbon dioxide via a biochemical process utilizing a cell extract containing an obligate or facultative oxyhydrogen microorganism and / or enzymes derived from an oxyhydrogen microorganism in one or more process steps, in certain embodiments. The invention also describes the fixation of inorganic carbon sources other than CO2 and / or other C1 carbon sources. Cell extracts include, but are not limited to, chemosynthetic enzyme-containing lysates, extracts, fractions, or purified products that can be produced from oxyhydrogen microorganisms by standard methods. In addition, the present invention provides compositions and methods for the recovery, treatment, and use of chemical products from one or more chemosynthetic reaction steps performed by oxyhydrogen microorganisms to fix inorganic carbon into organic compounds and / or from one or more synthetic reaction steps performed by oxyhydrogen microorganisms to extend C1 molecules into organic chemicals with longer carbon chains. Finally, in certain embodiments, the present invention provides compositions and methods for the generation, processing and delivery of chemical nutrients required for chemoautotrophic carbon fixation by oxyhydrogen microorganisms, specifically electron donors, including molecular hydrogen and / or electricity (but not limited thereto), and electron acceptors, including oxygen and carbon dioxide (but not limited thereto), to drive the carbon fixation reaction; compositions and methods for maintaining an environment conducive to carbon fixation by oxyhydrogen microorganisms; and compositions and methods for removing chemical products of chemosynthesis from an oxyhydrogen culture environment and recovering and recirculating unused chemical nutrients.

[0034] The terms "molecular hydrogen," "dihydrogen," and "H2" are used synonymously throughout this text.

[0035] The terms “oxyhydrogen microorganisms” and “hyperhydrogen microorganisms” are used synonymously throughout this text. Oxyhydrogen microorganisms are outlined in Chapter 5, Section III of “Thermophilic Bacteria” by Jakob Kristjansson, CRC Press, 1992 (which is incorporated herein by reference). Generally, oxyhydrogen microorganisms are capable of carrying out oxyhydrogen reactions. Oxyhydrogen microorganisms generally have the ability to use molecular hydrogen by utilizing hydrogenase, in which case some of the electrons donated from H2 become NAD. + The electrons are used for the reduction of (and / or other intracellular reducing equivalents), and the remaining electrons are used for aerobic respiration. In addition, oxyhydrogen microorganisms can generally autotrophically fix CO2 via pathways such as the reverse Calvin cycle and the reverse citric acid cycle.

[0036] In addition, the terms "oxyhydrogen reaction" and "detonating gas reaction" are used synonymously throughout to refer to the microbial oxidation of molecular hydrogen by molecular oxygen. An oxyhydrogen reaction is generally, 2H2 + O2 → 2H2O + energy This is expressed as and / or by the stoichiometric equivalence of this reaction.

[0037] Exemplary oxyhydrogen microorganisms usable in one or more process steps according to specific embodiments of the present invention include the following: purple non-sulfur photosynthetic bacteria (e.g., Rhodopseudomonas palustris, Rhodopseudomonas capsulata, Rhodopseudomonas viridis, Rhodopseudomonas sulfoviridis, Rhodopseudomonas blastica, Rhodopseudomonas spheroides, Rhodopseudomonas acidophila) Examples include, but are not limited to, Rhodopseudomonas acidophila, Rhodospirillum rubrum and other Rhodospirillum species, Rhodococcus opacus and other Rhodococcus species, Rhizobium japonicum and other Rhizobium species, Thiocapsa roseopersicina and other Thiocapsa species, Pseudomonas hydrogenovora, and Pseudomonas hydrogenothermophila. Hydrogenomonas (hydrogenomonas othermophila), and other species of the genus Pseudomonas, Hydrogenomonas pantotropha, Hydrogenomonas eutropha, Hydrogenomonas fasciis (HydrogenomonasHydrogenobacter thermophilus and other species of the genus Hydrogenobacter, Hydrogenovibrio marinus and other species of the genus Hydrogenovibrio, Helicobacter pylori and other species of the genus Helicobacter, Xanthobacter, Hydrogenophaga, Bradyrhizobium japonicum and other species of Bradyrhizobium, and Ralstonia eutropha. * eutropha* and other species of the genus Ralstonia, * Alcaligenes eutrophus* and other species of the genus Alcaligenes, * Variovorax paradoxus* and other species of the genus Variovorax, * Acidovorax facilis* and other species of the genus Acidovorax, cyanobacteria (e.g., Anabaena oscillarioides, Anabaena spiroides, Anabaena cylindrica, and other species of the genus Anabaena, but not limited to these), green algae (e.g., *Scenedesmus obliquus*) Chlamydomonas obliquus and other species of the genus Scenedesmus, Chlamydomonas reinhardiiThis includes, but is not limited to, species of the genus Chlamydomonas (reinhardii) and other species of the genus Chlamydomonas, species of the genus Ankistrodesmus, species of Rhaphidium polymorphium and other species of the genus Rhaphidium, and one or more consortia of microorganisms including oxyhydrogen microorganisms.

[0038] Various oxyhydrogen microorganisms usable in specific embodiments of the present invention may be native to a range of environments, including, but not limited to, hydrothermal vents, geothermal vents, hot springs, cold seeps, underground aquifers, salt lakes, salt layers, mines, acidic mine drainage, mine tailings, oil wells, refinery wastewater, contaminated water of oil, gas, or hydrocarbons, coal seams, deep underground formations, wastewater and sewage treatment plants, geothermal power plants, sulfurous vent areas, and soil (for example, soil contaminated with hydrocarbons, and / or soil located beneath or around oil or gas wells, oil refineries, oil pipelines, and gas stations). They may or may not be extremophiles, including, but not limited to, thermophiles, hyperthermophiles, acidophiles, halophiles, and psychrophiles.

[0039] In some embodiments, it is possible to produce chemical products with relatively long chains. For example, the organic chemicals produced in some embodiments may include compounds with carbon chain lengths of at least C5, at least C10, at least C15, at least C20, about C5 to about C30, about C10 to about C30, about C15 to about C30, or about C20 to about C30.

[0040] Figure 1 shows a general process flow diagram of an embodiment of the present invention, which comprises a process step for generating an electron donor (e.g., a molecular hydrogen electron donor) suitable for supporting chemosynthesis from energy input and crude inorganic chemical input; a step for recovering chemical by-products from the electron donor generation step; a step for delivering the generated electron donor, along with an oxygen electron acceptor, water, nutrients, and CO2 from an industrial flue gas point source, to one or more chemosynthesis reaction steps that utilize oxyhydrogen microorganisms to recover and fix carbon dioxide and produce chemicals and biomass by-products via a chemosynthesis reaction; a process step for recovering both chemicals and biomass products from the process stream; and a step for recirculating unused nutrients and process water, as well as cell aggregates required to maintain microbial culture, back to the carbon fixation reaction step.

[0041] In the embodiment illustrated in Figure 1, CO2-containing flue gas is recovered from a point source or emission source. Electron donors (e.g., H2) required for chemosynthesis can be generated from input inorganic chemicals and energy. The flue gas, along with electron donors and electron acceptors required to drive chemosynthesis, and a medium suitable for supporting microbial culture and carbon fixation via chemosynthesis, can be pumped into a bioreactor containing oxyhydrogen microorganisms. Cell cultures can be continuously inflowed into and out of the bioreactor. After the cell cultures leave the bioreactor, cell aggregates can be separated from the liquid medium. Cell aggregates required to replenish the cell culture population at a desired (e.g., optimal) level can be recycled and returned to the bioreactor. Excess cell aggregates can be dried to form a dried biomass product, which can be further processed into various chemicals, fuels, or nutrients. Following the cell separation step, extracellular chemical products of the chemosynthetic reaction can be removed and recovered from the process flow. Any undesirable waste products that may be present are then removed. After this, the liquid culture medium and any unused nutrients can be recycled and returned to the bioreactor.

[0042] Reduced inorganic chemicals that promote the growth of chemoautotrophs (e.g., H2, H2S, ferrous, ammonium, Mn 2+ Many of these can be readily produced using electrochemical and / or thermochemical processes known in the field of chemical industry, which can, in some cases, be powered by a variety of zero-carbon, low-carbon, and / or renewable power sources, including wind, hydroelectric, nuclear, photovoltaic, or solar thermal energy.

[0043] In certain embodiments of the present invention, a carbon-free or low-carbon and / or renewable power source is used to generate an electron donor, including, but not limited to, one or more of the following: photovoltaic, solar thermal, wind, hydroelectric, nuclear, geothermal, enhanced geothermal, ocean thermal, wave power, and tidal power. In certain embodiments of the present invention, oxyhydrogen microorganisms function as biocatalysts for converting renewable energy and / or low-carbon or carbon-free energy into liquid hydrocarbon fuels or generally high-energy-density organic compounds, using CO2 recovered from flue gas or from the atmosphere or ocean, which functions as a carbon source. These embodiments of the present invention can provide renewable energy technologies capable of producing transport fuels with significantly higher energy densities than when hydrogen gas is produced using a renewable energy source (which must be stored in a relatively heavy storage system (e.g., a tank or storage material)) or when it is used to charge a battery (which has a relatively low energy density). In addition, liquid hydrocarbon fuel products according to certain embodiments of the present invention may be more suitable for current transportation infrastructure compared to battery or hydrogen energy storage options.

[0044] The location of one or more process steps that generate an electron donor (e.g., a molecular hydrogen electron donor) in a general process flow of a particular embodiment of the present invention is illustrated by box 3 in Figure 1 (labeled "Electron Donor Generation"). Electron donors produced in specific embodiments of the present invention using electrochemical and / or thermochemical processes known in the field of chemical industry, and / or electron donors produced from natural sources, include molecular hydrogen and / or valence or conduction electrons in solid electrode materials and / or other reducing agents, including, but not limited to, one or more (but not limited to) of the following: ammonia, ammonium, carbon monoxide, dithionite, elemental sulfur, hydrocarbons, metabisulfite, nitrogen oxide, nitrite, sulfates such as thiosulfates (e.g., sodium thiosulfate (Na2S2O3) or calcium thiosulfate (CaS2O3)), sulfides such as hydrogen sulfide, sulfites, thionites, thionic acid, transition metals or their sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, sulfates, or carbonates in the soluble or solid phase.

[0045] In certain embodiments of the present invention, molecular hydrogen is used as the electron donor. The hydrogen electron donor is used in the following ways: electrolysis of water by methods (but not limited to) using proton exchange membranes (PEM), liquid electrolytes such as KOH, high-pressure electrolysis, and high-temperature electrolysis of steam (HTES); thermochemical decomposition of water by methods including the iron oxide cycle, cerium(IV) oxide-cerium(III) oxide cycle, zinc-zinc oxide cycle, sulfur-iodine cycle, copper-chlorine cycle, calcium-bromine-iron cycle, and hybrid sulfur cycle (but not limited to); electrolysis of hydrogen sulfide; thermochemical decomposition and / or electrochemical decomposition of hydrogen sulfide; methane reforming enabling carbon recovery and sequestration, coal gasification enabling carbon recovery and sequestration, the Kvaerner process and other processes producing carbon black products, gasification or pyrolysis of biomass enabling carbon recovery and sequestration, and ferrous (Fe)2+ ) from iron (Fe 3+ H23, which involves the oxidation of an electron source to a half-cell, including oxidation to or oxidation of sulfur compounds (in which case the oxidized iron or sulfur can be recycled back to a reduced state through additional chemical reactions with minerals, including metal sulfides, hydrogen sulfide, or hydrocarbons (but not limited to these)) (but not limited to these). + Hydrogen is produced by methods known in the field of chemical process industries, including, but not limited to, one or more other electrochemical or thermochemical processes known to produce hydrogen with low or no carbon dioxide emissions, such as the reduction of half-cells from to H2 (but not limited to these).

[0046] In certain embodiments of the present invention, the hydrogen electron donor is not necessarily produced with low or no carbon dioxide emissions, but the hydrogen is produced from waste energy sources or low-value energy sources using methods known in the art of the chemical process industry, including gasification, pyrolysis, or steam reforming (but not limited to) of supply materials such as municipal waste, black liquor, agricultural waste, wood waste, stranded natural gas, biogas, sour gas, methane hydrate, tires, sewage, fertilizer, straw, and low-value high-lignocellulose biomass in general (but not limited to these).

[0047] In certain embodiments of the present invention, which utilize molecular hydrogen as an electron donor for carbon fixation reactions carried out by oxyhydrogen microorganisms, there are chemical byproducts formed during the production of molecular hydrogen using renewable and / or CO2-free energy inputs. When water is used as the hydrogen source, oxygen may be a byproduct of water splitting through processes including, but not limited to, electrolysis or thermochemical water splitting. In certain embodiments of the present invention, which use water as the hydrogen source, some of the oxygen byproducts can be used in the oxyhydrogen carbon fixation process for the production of intracellular ATP via oxyhydrogen reactions enzymatically involved in oxidative phosphorylation. In certain embodiments of the present invention, oxygen produced by water splitting in excess of what is needed to maintain favorable (e.g., optimal) conditions for carbon fixation and organic compound production by oxyhydrogen microorganisms can be processed into a form suitable for sale through process steps known in the scientific and technical field of commercial oxygen gas production. In certain embodiments of the present invention, which use hydrogen sulfide as the hydrogen source, sulfur or sulfuric acid may be chemical byproducts of molecular hydrogen production. In certain embodiments of the present invention, which use sulfuric acid as a byproduct of hydrogen production, some of the sulfuric acid can be used for biomass hydrolysis in post-carbon fixation process steps. In certain embodiments of the present invention, any excess sulfuric acid and / or sulfur produced as a by-product (for example, in an amount exceeding the amount available elsewhere for use in the carbon recovery and conversion process according to a particular embodiment of the present invention) can be processed into a form suitable for sale through process steps known in the scientific and technical fields of commercial sulfuric acid and / or sulfur production. Process heat may also be generated during the production of hydrogen from hydrogen sulfide. In certain embodiments of the present invention, the process heat generated during hydrogen production is recovered and used elsewhere for the carbon recovery and conversion process according to a particular embodiment of the present invention to improve overall energy efficiency. Chemical by-products and / or thermal by-products and / or electrical by-products may accompany the production of molecular hydrogen for use as an electron donor in certain embodiments of the present invention. Chemical by-products and / or thermal by-products and / or electrical by-products of molecular hydrogen production can be used elsewhere for the carbon recovery and conversion process according to a particular embodiment of the present invention as much as possible to improve efficiency.In certain embodiments, additional chemical by-products (e.g., in amounts exceeding those usable in the carbon recovery and conversion process according to a particular embodiment of the present invention) can be prepared for sale to generate an additional revenue stream. Excess thermal or electrical energy by-products from the production of molecular hydrogen (e.g., in amounts exceeding those usable within the process) can be delivered for sale for use in other chemical and / or biological processes via means known in the science and technology of heat exchange and heat conduction, as well as electricity generation and electricity transfer, including, for example, (but not limited to) the conversion of process heat into electricity in a marketable form.

[0048] In certain embodiments of the present invention, electrochemical energy stored in solid valence electrons or solid conduction electrons within an electrode, capacitor, or associated device is utilized, either alone or in combination with a chemical electron donor and / or electron carrier, to provide a reduced equivalent for a carbon fixation reaction to an oxyhydrogen microorganism by direct exposure of the electrode material to a microbial culture environment and / or immersion of the electrode material in a microbial culture medium.

[0049] A feature of certain embodiments of the present invention relates to the generation or recycling of electron donors of mineral origin (for example, electron donors generated from minerals containing reduced forms of S and Fe) that can be similarly used by certain oxyhydrogen microorganisms as a reducing equivalent source in addition to or instead of hydrogen. Thus, in certain embodiments, the present invention may enable the use of largely undeveloped energy sources (inorganic geochemical energy).

[0050] The electron donor used in a particular embodiment of the present invention is the following: namely, elemental Fe 0 Siderite (FeCO3), magnetite (Fe3O4), pyrite or marcasite (FeS2), pyrrhotite (Fe (1-x)S(x = 0 to 0.2)), pentlandite (Fe,Ni)9S8, violarite (Ni2FeS4), pyrrhotite (Ni,Fe)S2, arsenopyrite (FeAsS), or other iron sulfides, realgar (AsS), orpiment (As2S3), cobaltite (CoAsS), rhodochrosite (MnCO3), chalcopyrite (CuFeS2), bornite (Cu5FeS4), covellite (CuS), tetrahedrite (Cu8Sb2S7), enargite (Cu3AsS4), arsenic tetrahedrite (Cu 12 As 4. S 13 ), chalcocite (Cu2S), or other copper sulfides, sphalerite (ZnS), marmatite (ZnS), or other zinc sulfides, galena (PbS), jamesonite (Pb5(Sb,As2)S8), or other lead sulfides, argentite or acanthite (Ag2S), molybdenite (MoS2), millerite (NiS), polydymite (Ni3S4), or other nickel sulfides, stibnite (Sb2S3), Ga2S3, CuSe, cooperite (PtS), laurite (RuS2), braggite (Pt,Pd,Ni)S, FeCl2, or one or more (but not limited to these) of natural mineral sources can be purified.

[0051] In certain embodiments of the present invention, the generation of electron donors from natural mineral sources may include a pretreatment step that includes (but is not limited to) increasing the leaching surface area by finely granulating, crushing, or grinding the mineral ore using a device such as a ball mill, and wetting the mineral ore to produce a slurry. In these embodiments of the present invention in which electron donors are generated from natural mineral sources, it may be advantageous to control the particle size so that sulfides and / or other reducing agents present in the ore can be concentrated by flotation methods such as dissolved air flotation or foam flotation using a flotation tower or mechanical flotation tank, gravity separation, magnetic separation, heavy liquid separation, selective flocculation, water separation, or fractional distillation (but is not limited to these). After the generation of crushed ore or slurry, particulate matter in the leaching or concentrate can be separated by filtration (e.g., vacuum filtration), sedimentation, or other well-known solid / liquid separation techniques before introducing the electron donor-containing solution into a chemical autotrophic culture environment. In addition, any toxic substances leached from the mineral ore to chemoautotrophs can be removed before exposing the leachate to chemoautotrophs. The solid remaining after processing the mineral ore can be concentrated by filter pressing, disposed of, retained for further processing, or sold, depending on the mineral ore used in the particular embodiment of the present invention.

[0052] Furthermore, in certain embodiments of the present invention, the electron donor may be any of the following: process gas, tail gas, fortified petroleum recovery vent gas, biogas, acidic mine wastewater, landfill leachate, landfill gas, geothermal gas, geothermal sludge or brine, metallic contaminants, gangue, tailings, sulfides, disulfides, mercaptans (e.g., methyl mercaptan and dimethyl mercaptan, ethyl mercaptan, etc.), carbonyl sulfides, carbon disulfides, alkanesulfonates, dialkyl sulfides, thiosulfates, thiofurans. It can be purified from contaminants or waste products, including but not limited to one or more of the following: thiocyanates, isothiocyanates, thioureas, thiols, thiophenols, thioethers, thiophenes, dibenzothiophenes, tetrathionates, dithionits, thionates, dialkyl disulfides, sulfones, sulfoxides, sulfolanes, sulfonic acids, dimethyl sulfoniopropionates, sulfonic acid esters, hydrogen sulfide, sulfate esters, organic sulfurs, sulfur dioxide, and all other sour gases.

[0053] In addition to mineral sources, electron donors, in certain embodiments of the present invention, may be of fossil origin but are generated or recycled through chemical reactions with hydrocarbons used in chemical reactions that result in low or no carbon dioxide emissions. These reactions include thermochemical and electrochemical processes. Such chemical reactions used in these embodiments of the present invention include, but are not limited to, sulfate reactions or thermochemical reductions of TSR and Muller-Kuhne reactions, and methane reforming-like reactions using metal oxides such as iron oxide, calcium oxide, or magnesium oxide (but not limited to these) instead of water (in which case the hydrocarbons are reacted with the hydrogen electron donor products to form solid carbonates with little or no carbon dioxide emissions).

[0054] Examples of reactions between metal oxides and hydrocarbons that produce hydrogen electron donor products and carbonates include: 2CH4 + Fe2O3 + 3H2O → 2FeCO3 + 7H2 and / or CH4 + CaO + 2H2O → CaCO3 + 4H2O These are some examples, but are not limited to them.

[0055] In certain embodiments, the generated electron donor is oxidized in one or more chemical synthesis steps by an electron acceptor, including, but not limited to, one or more valence band holes or conduction band holes in a solid electrode material, namely, carbon dioxide, oxygen, and / or ferric ions or other transition metal ions, nitrates, nitrites, sulfates, or valence band holes or conduction band holes in a solid electrode material.

[0056] The location of one or more chemical synthesis steps and / or oxyhydrogen reaction steps in a general process flow of a particular embodiment of the present invention is illustrated by box 4 in Figure 1, labeled "Bioreactor - Vapor Microorganism".

[0057] In each step of a process in which chemosynthesis and / or oxyhydrogen reactions are carried out, one or more types of electron donors and one or more types of electron acceptors are added to the nutrient medium containing oxyhydrogen microorganisms by bolus addition, periodic or continuous addition, pump-injection, or other means to the reaction vessel. Chemosynthetic reactions driven by the transfer of electrons from electron donors to electron acceptors can fix inorganic carbon dioxide into organic compounds and biomass.

[0058] In certain embodiments of the present invention, electron carriers can be incorporated into nutrient media to kinetically accelerate chemical synthesis reaction steps, thereby facilitating the delivery of reducing equivalents from electron donors to oxyhydrogen microorganisms in the presence of electron acceptors and inorganic carbon. This aspect of the present invention can be used to facilitate the transfer of reducing electrons from poorly soluble electron donors, such as electrons in H2 gas or solid electrode materials (but not limited to these), to oxyhydrogen microorganisms, using electron carriers known in the art of electrical stimulation of microbial metabolism, including but not limited to anthraquinone-2,6-disulfonate (AQDS), cobalt sepulcrate, cytochrome, formate, humic substances, iron, methyl viologen, NAD+ / NADH, neutral red (NR), phenazine, and quinones.

[0059] The delivery of reduced equivalents from electron donors to oxyhydrogen microorganisms for one or more chemosynthetic reactions can be kinetically and / or thermodynamically facilitated by means including, in certain embodiments, introducing a hydrogen storage material that can also function as a solid carrier medium for microbial growth into the microbial culture environment to place the absorbed or adsorbed hydrogen electron donors in close proximity to the hydrogen-oxidizing chemoautotrophs, and / or directly introducing an electrode material (e.g., graphite, graphite felt, activated carbon, carbon nanofibers, conductive polymers, steel, iron, copper, titanium, lead, tin, palladium, platinum, platinum-clad titanium, other platinum-clad metals, transition metals, transition metal alloys, sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, phosphates, sulfates, and / or carbonates) that can also function as a solid carrier medium and an electron donor or electron acceptor source into the chemoautotrophic culture environment to place solid electrons in close proximity to the microorganisms (but not limited to these). Some such embodiments of the present invention may be useful for transferring reduced equivalents from poorly soluble electron donors, such as H2 gas or electrons in solid electrode materials (but not limited to these), to oxyhydrogen microorganisms.

[0060] The culture broth used in the chemical synthesis steps according to a particular embodiment of the present invention may be an aqueous solution containing suitable minerals, salts, vitamins, cofactors, buffers, and other components required for microbial growth, as known to those skilled in the art [Bailey and Ollis, Biochemical Engineering Fundamentals, 2nd ed; pp. 383-384 and 620-622; McGraw-Hill: New York (1986)]. These nutrients can be selected to maximize carbon fixation and promote carbon flow via enzymatic pathways to obtain the desired organic compounds. Selective growth environments, such as those used in the art for solid-state or non-aqueous fermentation, may be used in particular embodiments. In particular embodiments utilizing aqueous cultures, broths, brine, seawater, and / or water from other natural water bodies, or water from other unsuitable drinking water sources, they may be used if they are resistant to oxyhydrogen microorganisms.

[0061] In certain embodiments of the present invention, the biochemical pathway can be controlled and optimized to produce chemical products (e.g., target organic compounds) and / or biomass by maintaining specific growth conditions (e.g., trace amounts of micronutrients such as nitrogen, oxygen, phosphorus, sulfur, and inorganic ions, and, if present, any regulatory molecules not generally considered nutrients or energy sources). Depending on the embodiment of the present invention, the broth can be maintained under aerobic, microaerobic, anaerobic, and facultative conditions. A facultative environment is considered to have an aerobic upper layer and an anaerobic lower layer resulting from stratification of the water column.

[0062] In certain embodiments of the present invention, oxygen levels are controlled. Oxygen levels can be controlled, for example, to promote the production of target organic compounds by oxyhydrogen microorganisms via carbon fixation. One objective of controlling oxygen levels is, in certain embodiments, to control (e.g., optimize) the intracellular adenosine triphosphate (ATP) concentration via cellular reduction of oxygen and ATP production by oxidative phosphorylation. In some such embodiments, it may be desirable to maintain a sufficiently reducing environment so that the intracellular ratio of NADH (or NADPH) to NAD (or NADP) is relatively high while controlling the ATP concentration. In some embodiments, ATP levels are increased and / or optimized within oxyhydrogen microorganisms by means of one or more (but not limited to) of the following: cellular reduction of oxygen and / or other electron acceptors with sufficient oxidative intensity for ATP production via oxidative phosphorylation; direct introduction of ATP into the culture medium; and / or direct introduction of chemical analogs of ATP into the culture medium.

[0063] The reduction of oxygen by hydrogen in oxyhydrogen reactions is generally enzymatically involved in ATP production via oxidative phosphorylation in oxyhydrogen microorganisms. Oxyhydrogen reactions can act as an alternative to the light-dependent reactions of photosynthesis, producing both NADPH and ATP. Generally, in oxyhydrogen microorganisms, hydrogenases catalyze the reduction of NAD to NADH by hydrogen (or, alternatively, in some photosynthetic organisms capable of oxyhydrogen reactions, hydrogenases catalyze the reduction of ferredoxin by H2, and thereby reduce NADP to NADPH) [Chen, Gibbs, Plant Physiol. (1992) 100, 1361-1365]. NADH and / or NADPH can then be used as reducing agents in assimilation reactions or to produce ATP by reducing oxygen via oxidative phosphorylation [Bongers, J. Bacteriology, (Oct 1970) 145-151]. Therefore, the following light-dependent photosynthetic reactions: 2H2O + 2NADP ++2ADP+2Pi+Optical→2NADPH+2H + +2ATP+O2 Instead, 1 / 2O2+2NADP + +2ADP+2Pi+3H2→2NADPH+2H + +2ATP+2H2O The oxyhydrogen reaction can be carried out under dark conditions (for example, in the substantial absence of visible electromagnetic radiation), and if we assume that 2 ATP are produced for each H2 consumed, then hydrogen acts in place of photons [Bongers, J. Bacteriology, (Oct 1970) 145-151].

[0064] In certain embodiments of the present invention, the goal is to maintain high intracellular concentrations of ATP, and even NADH and / or NADPH, in order to promote carbon fixation and to drive assimilation and / or solvent production pathways that consume reducing equivalents as well as ATP and / or reduce the net ATP yield of chemosynthetic carbon fixation. Such biochemical pathways include, but are not limited to, fatty acid synthesis, the mevalonate pathway and terpenoid synthesis, the butanol pathway and 1-butanol synthesis, the acetolactic acid / α-ketovaleric acid pathway and 2-butanol synthesis, and the ethanol pathway. In some embodiments of the present invention, it is possible to determine a preferred oxygen level. That is, if the oxygen level is too low, intracellular ATP in the oxyhydrogen microorganism may be reduced to below the desired level, while if the oxygen level is too high, the NADH (or NADPH) to NAD (or NADP) ratio may be reduced to below the desired level.

[0065] In certain embodiments of the present invention, the application of oxyhydrogen reactions to produce ATP and NADH and / or NADPH used for carbon fixation and the synthesis of organic compounds may offer superior advantages over alternative methods using anaerobic biochemical pathways for carbon fixation, such as the Wood-Ljungdahl pathway or the methane production pathway. Carbon fixation via the Wood-Ljungdahl pathway or the methane production pathway generally produces C1 or C2 organic compounds, and it can be difficult to produce compounds greater than C4 through these pathways.

[0066] The Wood-Ljungdahl pathway is essentially capable of producing acetic acid, ethanol, butyric acid, and butanol, although butyric acid and butanol are generally by-products of H2 and CO2 gas fermentation, and chain lengths greater than C4 typically do not occur [Lynd, Zeikus, J. of Bacteriology (1983) 1415-1423; Eichler, Schink, Archives of Microbiology (1984) 140, 147-152]. The acetic acid production pathway, leading to acetic acid and butyric acid, generates net ATP, while the solvent production pathway, leading to ethanol and butanol, does not [Papoutsakis, Biotechnology & Bioengineering (1984) 26, 174-187; Heise, Muller, Gottschalk, J. of Bacteriology (1989) 5473-5478; Lee, Park, Jang, Nielsen, Kim, Jung, Biotechnology & Bioengineering (2008) 101, 2, 209-228]. Since ATP is required for cell maintenance, there will generally be a certain amount of relatively undesirable non-biofuel by-products (i.e., organic acids) from acetic acid-producing bacteria that fix carbon via the Wood-Ljungdahl pathway, resulting in the waste of reducing equivalents and carbon.

[0067] The synthesis of hydrocarbons with chain lengths greater than C4 is most commonly achieved biologically via fatty acid biosynthesis [Fischer, Klein-Marcuschamer, Stephanolpoulos, Metabolic Engineering (2008) 10, 295-304]. Unlike the solvent synthesis pathway derived from the Wood-Ljungdahl pathway, fatty acid synthesis is involved in net ATP consumption. For example, the net reaction for the synthesis of palmitic acid (C16) is given below, starting with acetyl-CoA in this example. 8-acetyl-CoA + 7ATP + H2O + 14NADPH + 14H + → Palmitic acid + 8CoA + 14NADP + +7ADP+7Pi

[0068] One of the challenges in using anaerobic pathways such as methane production and Wood-Ljungdahl to drive fatty acid synthesis is that the amount of ATP produced per H2 consumed is relatively small. For methane [Thauer, RK, Kaster, AK, Seedorf, H., Buckel, W. & Hedderich, R Methanogenic archaea: ecologically relevant differences in energy conservation. Nat Rev Microbiol 6, 579-591, doi:nrmicro1931[pii]] or acetic acid production, it is 1 ATP per 4 H2, and for butyric acid production, it is 1 ATP per 10 H2 [Papoutsakis, Biotechnology & Bioengineering (1984) 26, 174-187, Heise, Muller, Gottschalk, J. of Bacteriology (1989) 5473-5478, Lee, Park, Jang, Nielsen, Kim, Jung, Biotechnology & [Bioengineering (2008) 101, 2, 209-228]. In contrast, in oxyhydrogen reactions, hydrogen-assimilating oxyhydrogen microorganisms can produce up to 2 ATP per H2 consumed [Bongers, J. Bacteriology, (Oct 1970) 145-151]. In other words, oxyhydrogen microorganisms can produce more than eight times the amount of ATP per H2 consumed by methane-producing or acetic acid-producing microorganisms. Furthermore, the ATP production pathway via oxyhydrogen reactions produces water that can be easily incorporated into the process stream, rather than the relatively undesirable acetic acid or butyrate products of acid production, which can disrupt the system's pH and rise to biotoxic concentrations.

[0069] The highest energy density fuel actually achievable naturally via the Wood-Ljungdahl pathway by introducing inorganic carbon is generally ethanol at 30 MJ / kg, although butanol at 36.1 MJ / kg is also possible. The production of diesel fuel (46.2 MJ / kg) or JP-8 aviation fuel (43.15 MJ / kg) can generally be difficult and less efficient than using anaerobic pathways such as Wood-Ljungdahl, because the amount of H2 that needs to be consumed in the absolute anaerobic pathway per ATP produced for fatty acid synthesis increases. However, these high-density, infrastructure-compatible liquid fuels can be readily produced via fatty acid synthesis pathways driven by ATP, NADH, or NADPH produced by oxyhydrogen reactions.

[0070] The lipid content of biomass and the efficiency of the lipid biosynthesis pathway are two factors that can affect the overall efficiency of certain embodiments of the present invention in converting CO2 and other C1 compounds into long-chain compounds (e.g., infrastructure-compatible fuels). The lipid content of biomass can determine the proportion of carbon and reducing equivalents that direct the synthesis of fuel products, in contrast to other components of the biomass. The lipid content can determine the energy input from reducing equivalents recoverable in the final fuel product. Similarly, the efficiency of the metabolic pathway can determine the amount of reducing equivalents that must be consumed when converting CO2 and hydrogen into lipids along the lipid biosynthesis pathway. Many oxyhydrogen microorganisms include species with high lipid content and efficient pathways from H2 and CO2 to lipids. In certain embodiments of the present invention, high-lipid-content species such as Rhodococcus opacus (but not limited to this) that can have a lipid content of more than 70% are used. [Gouda, MK, Omar, SH, Chekroud, ZA & Nour Eldin, HMBioremediation of kerosene I: A case study in liquid media. Chemosphere 69, 1807-1814, doi:S0045-6535(07)00738-2, Waltermann, M., Luftmann, H., Baumeister, D., Kalscheuer, R. & Steinbuchel, A. Rhodococcus opacus strain PD630 as a new source of high-value single-cell oil? Isolation and characterization of triacylglycerols and other storage lipids. Microbiology 146(Pt 5), 1143-1149 (2000).] and / or species that utilize highly efficient metabolic pathways such as the reverse tricarboxylic acid cycle [i.e., the reverse citric acid cycle] (but not limited to this) for carbon fixation [Miura, A., Kameya, M., Arai, H., Ishii, M. & Igarashi, Y.][A soluble NADH-dependent fumarate reductase in the reductive tricarboxylic acid cycle of Hydrogenobacter thermophilus TK-6. J Bacteriol 190, 7170-7177, doi:JB.00747-08[pii]10.1128 / JB.00747-08(2008)., Shively, JM, van Keulen, G. & Meijer, WG Something from almost nothing: carbon dioxide fixation in chemoautotrophs. Annu Rev Microbiol 52, 191-230, doi:10.1146 / annurev.micro.52.1.191(1998).] is used. In terms of energy efficiency, the reverse tricarboxylic acid pathway may be a relatively favorable pathway. The synthesis of palmitic acid from H2 and CO2 is generally about 15% more efficient than palmitic acid synthesis in acetic acid-producing bacteria, in terms of the amount of reduced equivalents consumed, because oxyhydrogen microorganisms increase ATP production per unit of reduced equivalent consumed in the oxyhydrogen reaction.

[0071] The inorganic carbon source used in the chemical synthesis reaction process steps according to a particular embodiment of the present invention includes, but is not limited to, one or more of the following: carbon dioxide-containing gas streams, which may be pure substances or mixtures; liquefied CO2; dry ice; dissolved carbon dioxide, carbonate ions, or minerals of bicarbonate, carbonates, or bicarbonates in solutions, including aqueous solutions such as seawater. Carbon dioxide and / or other forms of inorganic carbon can be introduced into the carbon fixation process steps by bolus addition, periodic addition, or continuous addition to the nutrient medium in the reaction vessel. The organic compound containing only one carbon atom that can be used in the synthesis reaction process steps according to a particular embodiment of the present invention includes, but is not limited to, one or more of the following: carbon monoxide, methane, methanol, formate, formic acid, and / or mixtures containing C1 chemicals (for example, various syngas compositions produced from various fixed carbon supply materials that have been gasified or steam reformed).

[0072] In certain embodiments, an organic compound and / or electron donor containing only one carbon atom is produced via gasification and / or pyrolysis of biomass and / or other organic matter (e.g., biomass and / or other organic matter from waste sources or low-value sources) and supplied to an oxyhydrogen microbial culture as syngas. In this case, the hydrogen-to-carbon monoxide ratio in the syngas may or may not be adjusted via means such as a water-to-gas conversion reaction before the syngas is delivered to the microbial culture. In certain embodiments, an organic compound and / or electron donor containing only one carbon atom is produced via methane or natural gas (e.g., stranded natural gas or other natural gas that is burned or released into the atmosphere) or methane steam reforming from biogas or landfill gas and supplied to an oxyhydrogen microbial culture as syngas. In this case, the hydrogen-to-carbon monoxide ratio in the syngas may or may not be adjusted via means such as a water-to-gas conversion reaction before the syngas is delivered to the microbial culture.

[0073] In certain embodiments of the present invention, carbon dioxide-containing flue gas is recovered from the chimney at a temperature, pressure, and gas composition characteristic of untreated exhaust, and then directed into a reactor where carbon fixation takes place with minimal modification. In some embodiments where no impurities harmful to chemoautotrophs are present in the flue gas, modification of the flue gas upon entering the reactor can be limited to compression required to pump the gas into the reactor system and / or heat exchange required to lower the gas temperature to a temperature suitable for microorganisms.

[0074] Oxyhydrogen microorganisms generally have a higher oxygen tolerance, giving them a significant advantage over obligate anaerobic acetic acid-producing or methane-producing microorganisms in carbon recovery applications. Since industrial flue gas is one of the CO2 sources intended for specific embodiments of the present invention, the relatively higher oxygen tolerance of oxyhydrogen microorganisms compared to obligate anaerobic methane-producing or acetic acid-producing bacteria allows them to withstand the 2-6% O2 content typically found in flue gas.

[0075] In some embodiments (for example, well known in the field of carbon capture technology) in which carbon dioxide-containing flue gas is transported and introduced into a system to dissolve carbon dioxide in a solution, the scrubbed flue gas (generally containing mainly inert gases such as nitrogen) can be released into the atmosphere.

[0076] In certain embodiments of the present invention, the gases dissolved in a solution and supplied to the culture broth, or dissolved directly in the culture broth, include carbon dioxide and gaseous electron donors (e.g., hydrogen gas), but in certain embodiments of the present invention, other electron donors such as carbon monoxide and other components of syngas, hydrogen sulfide, and / or other sour gases (but not limited to these). It is also possible to maintain a controlled amount of oxygen in the culture broth according to some embodiments of the present invention, and in certain embodiments, the oxygen will be effectively dissolved in the solution supplied to the culture broth and / or dissolved directly in the culture broth.

[0077] The dissolution of oxygen, carbon dioxide, and / or electron donor gases (e.g., hydrogen and / or carbon monoxide) can be achieved in some embodiments of the present invention using compressor, flow meter, and / or flow valve mechanisms known to those skilled in the art of bioreactor-scale microbial culture, and can be supplied to one or more of the following mechanisms commonly used for pumping gas into a solution: sparging devices, diffusers (e.g., dome-shaped, tubular, disc-shaped, or donut-shaped geometric shapes, but not limited thereto), coarse bubble or microbubble aerators, and / or venturi devices (but not limited thereto). In certain embodiments of the present invention, surface aeration can also be performed using a paddle aerator or the like. In certain embodiments of the present invention, gas dissolution is facilitated by mechanical mixing using an impeller and / or turbine. In some embodiments, it is possible to reduce the bubble size using a hydraulic shear device.

[0078] In certain embodiments of the present invention, where it is necessary to effectively pump air or oxygen into the culture broth to maintain a favorable (e.g., optimal) oxygen load level, oxygen bubbles are injected into the broth with a diameter desirable (e.g., optimal) for mixing and oxygen transport. This has been shown to be 2 mm in certain embodiments [Environment Research Journal May / June 1999 pgs.307-315]. In certain aerobic embodiments of the present invention, a process of shearing the oxygen bubbles is used to achieve this bubble diameter, as described in U.S. Patent No. 7,332,077. In some embodiments, the bubbles have an average diameter of 7.5 mm or less, and slugging is avoided.

[0079] In certain embodiments of the present invention that utilize hydrogen as an electron donor, hydrogen gas is supplied to a chemoautotrophic culture vessel by bubbling it into the culture medium and / or by diffusing it through a membrane that is in contact with the culture medium but impermeable to it. The latter method is considered safer and may be preferred in many embodiments because hydrogen accumulating in the gas phase can form explosive conditions (the explosive hydrogen concentration range in air is 4-74.5%, which is avoidable in certain embodiments of the present invention). In some embodiments, the membrane is coated with a biofilm of oxyhydrogen microorganisms so that after passing through the membrane, the hydrogen must diffuse into the microorganisms.

[0080] Additional chemicals known in the art that are necessary or useful for the maintenance and growth of oxyhydrogen microorganisms can be added to the culture broth according to a particular embodiment of the present invention. These chemicals include nitrogen sources such as ammonia, ammonium (e.g., ammonium chloride (NH4Cl), ammonium sulfate ((NH4)2SO4)), nitrates (e.g., potassium nitrate (KNO3)), urea, or organic nitrogen sources such as phosphates (e.g., disodium phosphate (Na2HPO4), potassium phosphate (KH2PO4), phosphoric acid (H3PO4), potassium dithiophosphate (K3PS2O2), potassium orthophosphate (K3PO4), dipotassium phosphate (K2HPO4)), sulfates, yeast extracts, chelated iron, potassium (e.g., potassium phosphate (KH2PO4), potassium nitrate (KNO3), potassium iodide (KI), potassium bromide (KBr)), as well as other inorganic salts, minerals, and micronutrients (e.g., sodium chloride (NaCl), magnesium sulfate (MgSO47H2O) or magnesium chloride (MgCl2), calcium chloride (CaCl2) or calcium carbonate (CaCO3), manganese sulfate) (MnSO47H2O) or manganese chloride (MnCl2), ferric chloride (FeCl3), ferrous sulfate (FeSO47H2O) or ferrous chloride (FeCl24H2O), sodium bicarbonate (NaHCO3) or sodium carbonate (Na2CO3), zinc sulfate (ZnSO4) or zinc chloride (ZnCl2), ammonium molybdate (NH4MoO4) or sodium molybdate (Na2MoO42H2O), cuprous sulfate (CuSO4) or copper chloride (CuCl Examples include, but are not limited to, 22H2O, cobalt chloride (CoCl26H2O), aluminum chloride (AlCl3.6H2O), lithium chloride (LiCl), boric acid (H3BO3), nickel chloride (NiCl26H2O), tin chloride (SnCl2H2O), barium chloride (BaCl22H2O), copper selenite (CuSeO45H2O), or sodium selenite (Na2SeO3), sodium metavanadate (NaVO3), and chromium salts.In certain embodiments, mineral salt medium (MSM) formulated by Schlegel et al. may be used [Thermophilic bacteria, Jakob Kristjansson, Chapter 5, Section III, CRC Press, (1992)].

[0081] In certain embodiments, the concentrations of nutritional chemicals (e.g., electron donors and electron acceptors) are maintained at levels favorable to enhanced (e.g., maximum) carbon uptake and fixation and / or the formation of organic compounds (e.g., levels as close as possible to their respective optimal levels), which vary depending on the oxyhydrogen species used but can be determined by those skilled in the art of oxyhydrogen microbial culture, either through public knowledge or without excessive experimentation.

[0082] In certain embodiments of the present invention, along with nutrient levels, waste product levels, pH, temperature, salinity, dissolved oxygen and dissolved carbon dioxide, gas and liquid flow rates, stirring speed, and pressure in the microbial culture environment are controlled. Operating parameters affecting carbon fixation can be monitored using sensors (e.g., by measuring the concentration of electron donors / acceptors using dissolved oxygen probes and / or redox probes) and can be controlled manually or automatically based on feedback from sensors using devices including, but not limited to, operating valves, pumps, and stirrers. The temperature of the inflow broth and even the inflow gas can be adjusted by means such as, but not limited to, heat exchangers.

[0083] The dissolution of gases and nutrients necessary for maintaining oxyhydrogen cultures and promoting carbon fixation, as well as the removal of inhibitory waste products, can be facilitated by stirring the culture broth. Oxyhydrogen microorganisms can carry out carbon fixation reactions throughout the entire volume of the reactor, offering superior advantages over other methods, including those utilizing photosynthetic organisms (which are limited to surface areas because photosynthesis requires light). The use of stirring can further enhance this advantage by distributing the microorganisms, nutrients, optimal growth environment, and / or CO2 as broadly and uniformly as possible throughout the reactor volume to promote production (for example, so that the reactor volume in which carbon fixation reactions occur at the optimal rate is maximized).

[0084] In certain embodiments of the present invention, stirring of the culture broth can be achieved by recirculating the broth from the bottom to the top of the container via a recirculation conduit, sparging with carbon dioxide, an electron donor gas (e.g., H2), oxygen, and / or air, and / or by a device including a mechanical mixer (but not limited to) such as an impeller (100-1000 rpm) or a turbine (but not limited to).

[0085] In certain embodiments of the present invention, the chemical environment, oxyhydrogen microorganisms, electron donors, electron acceptors, oxygen, pH, and / or temperature levels are varied spatially and / or transiently across a series of fluid-connected bioreactors so that a wide variety of carbon fixation reactions and / or biochemical pathways leading to organic compounds occur sequentially or in parallel.

[0086] In certain embodiments of the present invention, the nutrient medium containing oxyhydrogen microorganisms can be partially or completely removed from the bioreactor periodically or continuously, and can be replaced with new cell-free medium for, for example, to maintain the cell culture in the exponential growth phase, to maintain the cell culture in the growth phase (exponential or quiescent phase) with an increased (e.g., optimal) carbon fixation rate, to replenish depleted nutrients in the growth medium, and / or to remove inhibitory waste products.

[0087] Because oxyhydrogen species can achieve high growth rates, it is possible to match or exceed the highest rates of carbon fixation and / or biomass production per unit biomass achievable by photosynthetic microorganisms. As a result, in certain embodiments, it is possible to generate excess biomass. Excess cell aggregates can be removed from the system to produce biomass products. In some embodiments, excess cell aggregates can be removed from the system to maintain a desirable (e.g., optimal) microbial population and cell density in the microbial culture so that a continuously high carbon recovery and fixation rate can be obtained.

[0088] Another advantage of certain embodiments of the present invention relates to a vessel used to contain and culture a carbon fixation reaction environment in a carbon recovery and fixation process. Exemplary culture vessels usable in some embodiments of the present invention for culturing and growing oxyhydrogen microorganisms for carbon dioxide recovery and fixation include those known to those skilled in the art of large-scale microbial culture. Such culture vessels, whether of natural or artificial origin, include, but are not limited to, airlift reactors, biological scrubber towers, bioreactors, bubble towers, cabarns, caves, cisterns, continuous agitated tank reactors, countercurrent upward expansion bed reactors, digesters, and specifically, digester systems known in the prior art of sewage / wastewater treatment or bioremediation, filters, such as trickle filters, rotating biological contactor filters, rotating disks, soil filters (but not limited to these), fluidized bed reactors, gaslift fermenters, immobilized cell reactors, lagoons, membrane biofilm reactors, microbial fuel cells, ore shafts, Pachuca tanks, packed bed reactors, plug-in flow reactors, ponds, pools, quarries, storage tanks, static mixers, tanks, towers, flowbed reactors, bats, vertical shaft bioreactors, and wells. The tank bottom, siding, walls, lining, and / or top can be constructed from one or more materials, including but not limited to bitumen, cement, ceramics, clay, concrete, epoxy, fiberglass, glass, macadam, plastic, sand, sealant, soil, steel or other metals and their alloys, stone, tar, wood, and any combination thereof. In certain embodiments of the present invention where oxyhydrogen microorganisms require a corrosive growth environment and / or produce corrosive chemicals via carbon fixation reactions, corrosion-resistant materials can be used to line the inside of the container that comes into contact with the growth medium.

[0089] Because oxyhydrogen microorganisms do not require sunlight to fix CO2, they can be used in carbon capture and fixation processes that avoid many of the drawbacks that can be associated with photosynthesis-based carbon capture and conversion technologies. For example, shallow, wide ponds are not required to maintain chemosynthesis, nor are bioreactors with high surface area-to-volume ratios and special functions or transparent materials like solar thermal collectors. Technologies such as certain embodiments of the present invention using oxyhydrogen microorganisms are not subject to the circadian, geographical, meteorological, and seasonal constraints typically associated with photosynthesis-based systems.

[0090] In certain embodiments of the present invention, by using chemical synthesis vessel shapes with a low surface area-to-volume ratio, such as cubic, cylindrical, ellipsoidal, or "egg-shaped," hemispherical, or spherical shapes (but not limited to these), material costs can be minimized unless other design requirements (e.g., land area) take precedence over material costs. In contrast to photosynthesis technologies, where a large surface area-to-volume ratio is required to provide sufficient light irradiation, the use of small reactor shapes may be due to the absence of photoreactivity in the chemical synthesis reaction.

[0091] Furthermore, since oxyhydrogen microorganisms are not light-dependent, it becomes possible to design plants with a much smaller footprint than those traditionally associated with photosynthesis. For example, if it is anticipated that the plant footprint must be minimized due to land use limitations, a long vertical shaft bioreactor system can be used for chemosynthetic carbon recovery. Long vertical shaft bioreactors are described, for example, in U.S. Patents No. 4,279,754, No. 5,645,726, No. 5,650,070, and No. 7,332,077.

[0092] Unless other requirements take precedence, in certain embodiments of the present invention, the introduction of erosive material into the reactor and / or the surface of the tank, which causes high losses of water, nutrients, and / or heat, is minimized. The ability to minimize such surfaces may be due to the absence of photonability in chemosynthesis. Surfaces that cause high losses of water, nutrients, and / or heat, and are also lost by erosion, are generally the same surfaces that transmit the light energy necessary for photosynthesis, so in general, photosynthesis-based technologies cannot minimize such surfaces.

[0093] In some embodiments of the present invention, the culture vessels can utilize reactor designs known to those skilled in the art of large-scale microbial culture to maintain an aerobic, microaerobic, anaerobic, anaerobic, or facultative environment, depending on the embodiment of the invention. For example, similar to the designs of many wastewater treatment plants, in certain embodiments of the invention, to carry out a number of chemosynthetic process steps, and in certain embodiments heterotrophic process steps, on a carbon dioxide waste stream, tanks are arranged sequentially in series forward fluid communication, with certain tanks maintained under aerobic conditions and others under anaerobic conditions.

[0094] In certain embodiments of the present invention, oxyhydrogen microorganisms are immobilized within their growth environment. Microbial immobilization can be achieved using any culture medium known in the art of microbial culture that supports colonization by microorganisms, for example, using a culture medium for growing microorganisms on a matrix, mesh, or membrane made from a wide range of natural and synthetic materials and polymers, including, but not limited to, one or more of the following: glass wool, clay, concrete, wood fibers, inorganic oxides (e.g., ZrO2, Sb2O3, or Al2O3), organic polymer polysulfone, or open-porous polyurethane foam having a high specific surface area. Furthermore, microorganisms in specific embodiments of the present invention can be grown on the surface of unbound objects distributed throughout the growth vessel as known in the art of microbial culture, such as, but are not limited to, beads, sand, silicates, meerschmitt, glass, ceramics, small-diameter plastic discs, spheres, tubes, particles, or other shapes known in the art, shredded coconut husk, crushed corn cob, activated carbon, granular coal, crushed coral, sponge balls, suspension media, small-diameter rubbery (elastomeric) polyethylene tubes, porous cloth, Berl saddles, and Raschig ring suspension strings (but are not limited to these). The materials used in the microbial support medium may include hydrogen storage materials and / or electrode materials to increase the transfer of reduction equivalents to oxyhydrogen microorganisms. Suitable electrode materials include, but are not limited to, one or more of the following: graphite, activated carbon, carbon nanofibers, conductive polymers, steel, iron, copper, titanium, lead, tin, palladium, platinum, transition metals, transition metal alloys, transition metal sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, phosphates, sulfates, or carbonates of transition metals.Examples of hydrogen storage materials usable for this application include, but are not limited to, titanium, graphite, activated carbon, carbon nanofibers, iron, copper, lead, tin, metal hydrides (e.g., TiFeH2, TiH2, VH2, ZrH2, NiH, NbH2, PdH), polymers known in the field of hydrogen storage (e.g., but are not limited to metal-organic frameworks (MOFs)), and nanoporous polymer materials. In certain embodiments, the hydrogen storage material does not react strongly with water and does not have a strong or rapid effect on the pH of the culture medium.

[0095] Inoculation of oxyhydrogen cultures into culture tanks can be carried out by methods including, but are not limited to, the transfer of cultures from existing oxyhydrogen cultures residing in other carbon recovery and fixation systems according to specific embodiments of the present invention and / or the transfer of cultures from incubation of seed stocks induced in an incubator. Seed stocks of oxyhydrogen strains can be transported and stored in forms including powder, liquid, frozen, or lyophilized, and any other suitable form readily apparent to those skilled in the art (but are not limited to these). In specific embodiments where cultures are established in very large reactors, culture growth and establishment can be carried out by sequentially increasing the size in intermediate-scale containers before inoculation into phenyl-scale tanks.

[0096] The location of one or more process steps for separating cell aggregates from the process stream in a general process flow of a particular embodiment of the present invention is illustrated by box 5 in Figure 1, labeled “Cell Separation”.

[0097] The separation of cell aggregates from a liquid suspension can be carried out by methods known in the art of microbial culture, including, but not limited to, one or more of the following: centrifugation, flocculation, flotation, filtration using membrane, hollow fiber, vortex, or ceramic filter systems, vacuum filtration, tangential fluid filtration, clarification, sedimentation, and liquid cyclone. [Examples of cell aggregate collection techniques are given in International Publication No. 08 / 00558, published 8 January 1998, U.S. Patent Nos. 5,807,722, 5,593,886, and 5,821,111]. In certain embodiments where the cell aggregates are fixed on a matrix, they can be collected by methods including gravity sedimentation or filtration (but not limited to) and separated from the growth substrate by liquid shear force.

[0098] In certain embodiments of the present invention, when excess cell aggregates are removed from the culture, they can be recirculated back into the cell culture in combination with a new broth, as indicated by the process arrow labeled “Recirculated Cell Aggregates” in Figure 1, so that sufficient biomass is retained in one or more chemosynthetic reaction steps. This allows for the continuous enhancement of (e.g., optimal) autotrophic carbon fixation by organic compounds. The cell aggregates recovered by the collection system can be recirculated back into the culture vessel, for example, using an airlift pump or intermittent pump. In certain embodiments, the cell aggregates recirculated back into the culture vessel are not exposed to flocculants, provided that the flocculants are non-toxic to microorganisms.

[0099] In certain embodiments of the present invention, microbial culture and carbon fixation reactions are maintained in a steady state over time at a target level, where the cell population and environmental parameters (e.g., cell density, chemical concentration) are controlled by continuous inflow and removal of nutrient medium and / or biomass. Cell density can be monitored in certain embodiments of the present invention by direct sampling, by correlation between optical density and cell density, and / or by a particle size analyzer. The hydraulic pressure holding time and biomass holding time can be separated to allow independent control of both broth chemistry and cell density. The hydraulic pressure holding time can be kept relatively short compared to the biomass holding time, and the dilution ratio can be kept high enough to ensure a large amount of broth is replenished for cell growth. The dilution ratio is set to an optimal trade-off between replenishment of culture broth and the increased process costs resulting from pump injection, increased input volume, and other requirements associated with the dilution ratio.

[0100] To support the processing of biomass products into biofuels or other useful products, after the cell recirculation process, it is possible to destroy and release excess microbial cells in certain embodiments of the present invention using methods including, for example, ball milling, cavitation pressure, sonication, or mechanical shearing (but not limited to these).

[0101] The biomass collected in some embodiments can be dried in one or more process steps in box 7 labeled “Dryer” in the general process flow of a particular embodiment of the present invention illustrated in Figure 1.

[0102] In certain embodiments of the present invention, drying of excess biomass can be carried out using techniques including, but not limited to, centrifugal separation, drum drying, evaporation, freeze-drying, heating, spray drying, vacuum drying, and / or vacuum filtration. Waste heat from industrial sources of flue gas can be used for drying biomass in certain embodiments. In addition, the chemical synthesis and oxidation of electron donors is generally exothermic and generally generates waste heat. In certain embodiments of the present invention, waste heat can be used for drying biomass.

[0103] In certain embodiments of the present invention, biomass is further treated after drying to support the production of biofuels or other useful chemicals by separating lipid content or other target biochemicals from microbial biomass. Separation of lipids can be carried out using nonpolar solvents for lipid extraction, such as hexane, cyclohexane, ethyl ether, alcohols (isopropanol, ethanol, etc.), tributyl phosphate, supercritical carbon dioxide, trioctylphosphine oxide, secondary and tertiary amines, or propane (but not limited to these). Other useful biochemicals can be extracted using solvents including chloroform, acetone, ethyl acetate, and tetrachloroethylene (but not limited to these).

[0104] The extracted lipid content of biomass can be processed using methods known in the field of biomass refining, including, but not limited to, one or more of the following: catalytic cracking and reforming, decarboxylation, hydrogenation, and isomerization, to produce petroleum and petrochemical substitutes, including, but not limited to, one or more of the following: JP-8 jet fuel, diesel oil, gasoline, and other alkanes, olefins, and aromatic compounds. In some embodiments, the extracted lipid content of biomass can be converted into ester-based fuels such as biodiesel oil (fatty acid methyl ester or fatty acid ethyl ester) via processes known in the field of biomass refining, including, but not limited to, transesterification and esterification.

[0105] The broth remaining after the removal of cell aggregates can be pumped into a system to remove chemical products of chemosynthesis and / or used nutrients that are recycled or recovered and / or discarded as much as possible.

[0106] In a typical process flow of a particular embodiment of the present invention, the location of one or more process steps for recovering chemicals from the process stream is illustrated by box 8 in Figure 1, labeled “Separation of Chemical By-products”.

[0107] The recovery and / or recycling of chemosynthetic chemical products and / or spent nutrients from aqueous broth solutions is achievable in certain embodiments of the present invention using apparatus and techniques known in the art of process industries and targeted to obtain the chemical products of certain embodiments of the present invention, for example, using solvent extraction, water extraction, distillation, fractional distillation, cementation, chemical precipitation, alkaline solution absorption, absorption or adsorption onto activated carbon, ion exchange resin, or molecular sieve, modification of solution pH and / or oxidation-reduction potential, evaporators, fractional crystallizers, solid / liquid separators, nanofiltration, and all combinations thereof (but not limited to these).

[0108] In certain embodiments of the present invention, long-chain organic compounds suitable for purification into free fatty acids, lipids, or other media, or into biofuel products produced via chemosynthesis, can be recovered from the process stream in the step shown in box 8 of Figure 1. These free organic molecules can be released into the process stream solution from oxyhydrogen microorganisms by means including, but not limited to, cellular excretion or secretion or cytolysis. In certain embodiments of the present invention, the recovered organic compounds are treated using methods known in the scientific and technical field of biomass purification, including, but not limited to, one or more of catalytic cracking and reforming, decarboxylation, hydrogenation, and isomerization. Such processes can be used to produce petroleum and petrochemical substitutes, including, but not limited to, JP-8 jet fuel, diesel oil, gasoline, and one or more of, but not limited to, other alkanes, olefins, and aromatic compounds. The recovered fatty acids can be converted into ester-based fuels such as biodiesel oil (fatty acid methyl esters or fatty acid ethyl esters) via processes known in the scientific and technical field of biomass purification, including, but not limited to, transesterification and esterification.

[0109] In some embodiments, after the recovery of chemicals from the process stream, the removal of waste products is carried out as shown by box 9 labeled “Waste Removal” in Figure 1. The residual broth can be returned to the culture vessel along with the replacement water and / or replacement nutrients.

[0110] In certain embodiments of the present invention, which involve the chemoautotrophic oxidation of electron donors extracted from mineral ores, solutions of metal oxide cations may remain after the chemical synthesis reaction steps. Furthermore, solutions rich in dissolved metal cations may, in particular, result from the introduction of contaminated flue gases from coal-fired plants or the like into the process. In some such embodiments of the present invention, metal cations can be stripped from the process stream by methods including, but are not limited to, cementation of scrap iron, cotton, copper, or zinc powder; chemical precipitation as a precipitate of sulfides or hydroxides; electrolytic extraction for plating specific metals; absorption onto activated carbon or ion-exchange resins; modification of solution pH and / or oxidation-reduction potential; and solvent extraction. In certain embodiments of the present invention, the recovered metals can be sold for use in additional revenue streams.

[0111] In certain embodiments, the chemicals used in the processes of recovering chemical products, recycling nutrients and water, and removing waste are low in toxicity to humans and low in toxicity to the oxyhydrogen microorganisms used when exposed to the process stream that is recycled and returned to the growth vessel.

[0112] In certain embodiments of the present invention, the pH of the microbial culture is controlled. To address a decrease in pH, a neutralization step can be performed to maintain the pH within an optimal range for the maintenance and growth of microorganisms before recirculating the broth back into the culture vessel. Neutralization of acids in the broth can be achieved by adding bases, including but not limited to limestone, lime, sodium hydroxide, ammonia, potassium hydroxide, magnesium oxide, and iron oxide. In certain embodiments, the bases are produced from carbon dioxide-free sources such as naturally occurring basic minerals, including but not limited to calcium oxide, magnesium oxide, iron oxide, iron ore, metal oxide-containing olivine, metal oxide-containing serpentine, metal oxide-containing ultrabasic sediments, and subsurface basic salt aquifers. When limestone is used for neutralization, carbon dioxide will generally be released. This carbon dioxide can be directed back into the growth vessel for chemosynthesis and / or sequestered in some other way rather than being released into the atmosphere.

[0113] A further feature of a particular embodiment of the present invention relates to the use of organic compounds and / or biomass produced through one or more chemical synthesis process steps according to a particular embodiment of the present invention. The use of the produced organic compounds and / or biomass includes the production of liquid fuels such as JP-8 jet fuel, diesel oil, gasoline, octane, biodiesel oil, butanol, ethanol, propanol, isopropanol, propane, alkanes, olefins, aromatic compounds, fatty alcohols, fatty acid esters, alcohols (but not limited to these), 1,3-propanediol, 1,3-butadiene, 1,4-Butanediol, 3-Hydroxypropionate, 7-ADCA / Cephalosporine, ε-Caprolactone, γ-Valerolactone, Acrylate, Acrylic Acid, Adipic Acid, Ascorbate, Aspartate, Ascorbic Acid, Aspartic Acid, Caprolactam, Carotenoids, Citrate, Citric Acid, DHA, Docetaxel, Erythromycin, Ethylene, Gamma-Butyrolactone, Glutamate, Glutamic Acid, HPA, Hydroxybutyrate, Isopentenol, Isoprene, Isoprenoids, Itaconate, Itaconic Acid, Lactate, Lactic Acid, Lanosterol, Levulinic Acid, Lycopene, Lysine, Malate, Malonic Acid, Peptide, ω-3DHA, ω-Fatty Acid, Paclitaxel, PHA Organic chemicals, including PHB, polyketides, polyols, propylene, pyrrolidones, serine, sorbitol, statins, steroids, succinates, terephthalates, terpenes, THF, rubber, wax esters, polymers, general chemicals, industrial chemicals, specialty chemicals, paraffin substitutes, additives, nutritional supplements, nutritional drugs, pharmaceuticals, pharmaceutical intermediates, and personal care products (but not limited to these), are used as raw materials and / or feedstocks for the production, manufacturing processes, or chemical processes of organic chemicals, including alcohol fermentation or other biofuel fermentation and / or gasification and liquefaction processes and / or other biofuel production processes (e.g., catalytic cracking, direct liquefaction, Fisher As a source of feedstock for the Tropsch process, hydrogenation, methanol synthesis, pyrolysis, transesterification, or microbial syngas conversion (including, but not limited to, these processes), as biomass fuel for combustion, specifically as fuel for co-combustion with fossil fuels, as a source of pharmaceuticals, medicinal substances, or nutrients, commercially available enzymes, antibiotics, amino acids, vitamins, bioplastics, glycerol, or 1,Its uses include, but are not limited to, as a carbon source for large-scale fermentation producing various chemicals including 3-propanediol (but not limited to these), as a nutrient source for the growth of other microorganisms or organisms, as feed for animals including cattle, sheep, chickens, pigs, or fish (but not limited to these), as a raw material for the production of methane or biogas, and as a fertilizer, soil additive, and soil stabilizer.

[0114] A further feature of certain embodiments of the present invention relates to the optimization of oxyhydrogen microorganisms for carbon dioxide capture, carbon fixation to organic compounds, and the production of other valuable chemical byproducts. This optimization can be carried out by methods known in the art of artificial breeding, including but not limited to accelerated mutagenesis (e.g., using ultraviolet light treatment or chemical treatment), genetic engineering or genetic modification, hybridization, synthetic biology, or traditional selective breeding. In certain embodiments of the present invention utilizing a microbial consortium, the community can be enriched with desired oxyhydrogen microorganisms by methods known in the art of microbiology, through growth in the presence of a target electron donor, including but not limited to hydrogen, an acceptor, including but not limited to oxygen, and environmental conditions.

[0115] A further feature of certain embodiments of the present invention relates to the modification of the biochemical pathway of oxyhydrogen microorganisms for producing target organic compounds. This modification can be achieved by manipulating the growth environment and / or by methods known in the art of artificial breeding, including but not limited to accelerated mutagenesis (e.g., using ultraviolet light treatment or chemical treatment), genetic engineering or genetic modification, hybridization, synthetic biology, or traditional selective breeding. The organic compounds produced through the modification include, but are not limited to, one or more of the following: biofuels, including but not limited to JP-8 jet fuel, diesel oil, gasoline, biodiesel oil, butanol, ethanol, long-chain hydrocarbons, lipids, fatty acids, pseudo-vegetable oils, and methane produced in vivo from biological reactions, or organic compounds and / or biomass optimized as feedstock for producing biofuels and / or liquid fuels via chemical post-treatment. These forms of fuels can be used as renewable / alternative energy sources with low-temperature room effect gas emissions.

[0116] To provide a specific example of the entire biochemical process for recovering CO2 and generating biomass and other useful by-products using oxyhydrogen microorganisms, a process flow diagram describing a specific embodiment of the present invention is provided and explained below. This specific example is not to be considered as limiting the present invention in any way, but is provided solely for illustrative purposes.

[0117] Figure 2 includes an exemplary process flow diagram illustrating one embodiment of the present invention for CO2 recovery and production of lipid-rich biomass (which is converted into JP-8 jet fuel) using oxyhydrogen microorganisms. In this group of embodiments, carbon dioxide-rich flue gas is recovered from emission sources such as power plants, refineries, or cement manufacturers. Next, the flue gas can be compressed and pumped into a cylindrical anaerobic digester containing one or more oxyhydrogen microorganisms, including Rhodopseudomonas palustris, Rhodopseudomonas capsulata, Rhodopseudomonas viridis, Rhodopseudomonas sulfoviridis, Rhodopseudomonas blastica, and other species of the genus Rhodopseudomonas (but not limited to these), as well as purple non-sulfur photosynthetic bacteria (but not limited to these).

[0118] In some embodiments, Rhodopseudomonas capsulata can be used as the acid-hydrogen microorganism, and in some cases, a doubling time of 6 hours can be achieved by chemoautotrophic growth with hydrogen. See, for example, Madigan, Gest, J. Bacteriology (1979) 524-530 (as incorporated herein by reference). In some embodiments, the doubling time of the microorganism may be 6 hours or less. In some embodiments, the dry biomass concentration may be at least about 3 g / l, at least about 4 g / l, or at least 5 g / l at a steady state. In some embodiments, the lipid content of the biomass in the acid-hydrogen microorganism may be at least about 10%, at least about 20%, at least about 30%, at least about 35%, or at least about 40%. For example, in some embodiments, Rhodopseudomonas palustris can be used as the acid-hydrogen microorganism. See, for example, Carlozzi, Pintucci, Piccardi, Buccioni, Minieri, Lambardi, Biotechnol. Lett., (2009) DOI 10.1007 / s10529-009-0183-2 (as incorporated herein by reference). In certain embodiments, the lipid content of the oxyhydrogen microbial biomass is at least 40%, a steady-state bioreactor cell density of at least 5 g / liter is present in a continuous process, the microbial doubling time is up to 6 hours, the process achieves an energy efficiency of at least 40% in the conversion from hydrogen to biomass, and / or at least 60% of the biomass energy content is stored as lipids (this corresponds to a lipid content of approximately 40 wt% of the biomass).

[0119] In a group of embodiments shown in Figure 2, hydrogen electron donors, as well as oxygen and carbon dioxide electron acceptors, are continuously added to the growth broth along with other nutrients required for chemosynthesis and the maintenance and growth of the culture, which are pumped into a digester. In certain embodiments, the hydrogen source is a carbon dioxide-free process. Exemplary carbon dioxide-free processes include electrolytic or thermochemical processes powered by energy technologies such as, for example, photovoltaic, solar, wind, hydroelectric, nuclear, geothermal, enhanced geothermal, ocean thermal, wave, and tidal power. In a group of embodiments shown in Figure 2, oxygen functions as an electron acceptor in the chemosynthetic reaction of intracellular ATP production via oxyhydrogen reactions associated with oxidative phosphorylation. Oxygen can be generated from flue gas, it can be produced from water splitting reactions used for hydrogen production, and / or it can be extracted from air. In Figure 2, carbon dioxide from flue gas is used for ATP produced via oxyhydrogen reactions and NAD by H2. + or NADP +NADH and / or NADPH, produced from intracellular enzyme-catalyzed reduction, function as electron acceptors for the synthesis of organic compounds via biochemical pathways utilizing these acceptors. The cell aggregate can be separated from the culture broth by continuously removing it from the digester and flowing it through a membrane filter. The cell aggregate can then be recycled back into the digester and / or pumped into the post-treatment for lipid extraction according to methods known to those skilled in the art. The lipids can then be converted into JP-8 jet fuel using methods known to those skilled in the art of biomass purification (see, for example, the USDOE Energy Efficiency & Renewable Energy Biomass Program, “National Algal Biofuels Technology Roadmap”, May 2010 (which is incorporated herein by reference in its entirety)). The cell-free broth, after passing through the cell aggregate removal filter, can then be subjected to any additional waste removal treatments as needed, depending on the flue gas source. The residual water and nutrients can then be pumped back into the digester.

[0120] Some species of the genus Rhodopseudomonas possess extremely versatile metabolisms, enabling photoautotrophic, photoheterotrophic, heterotrophic, and even chemoautotrophic growth, and allowing them to survive in both aerobic and anaerobic environments [Madigan, Gest, J. Bacteriology (1979) 524-530]. In specific embodiments of the present invention, the heterotrophic function of Rhodopseudomonas species is utilized to further improve the efficiency of carbon conversion to energy and lipid products. The non-lipid biomass residue after lipid extraction consists mainly of proteins and carbohydrates. In a particular embodiment of the present invention, as shown in Figure 2, a portion of the carbohydrate residue and / or protein residue after lipid extraction is acid hydrolyzed into monosaccharides and / or amino acids, the acid is neutralized, and the solution of monosaccharides and / or amino acids is supplied to a second heterotrophic bioreactor containing a species of the genus Rhodopseudomonas that consumes biomass input to produce additional lipid products.

[0121] The genome of Rhodopseudomonas palustris was sequenced by the DOE Joint Genome Institute [Larimer et al. (2003) Nature Biotechnology 22, 55-61]. Its genetic system has been reported to be particularly easy to modify. In a group of embodiments of the present invention, one or more carbon fixation reactions are carried out by species of the genus Rhodopseudomonas that have been modified, optimized, or engineered to perform improved fixation of carbon dioxide and / or other forms of inorganic carbon and / or improved production of organic compounds, by methods including, but not limited to, one or more of the following: accelerated mutagenesis, genetic engineering or genetic modification, hybridization, synthetic biology, or traditional selective breeding.

[0122] Figure 3 includes an exemplary schematic diagram of a bioreactor 300 that can be used in a particular embodiment. The bioreactor 300 can be used, for example, as the reactor exemplified as box 4 in Figure 1 labeled "Bioreactor - Oxygen Microorganisms" and / or as the reactor exemplified as box 4 in Figure 2 labeled "Bioreactor - Purple Non-Sulfur Bacteria". The bioreactor 300 exemplified in Figure 3 can be operated to take advantage of the low solubility of hydrogen and oxygen gases in water and to avoid dangerous mixing of hydrogen and oxygen gases. In addition, the bioreactor can provide oxygen and hydrogen required for cellular energy and carbon fixation to oxyhydrogen microorganisms by, for example, sparging, bubbling, or diffusing oxygen or air upward into a vertical liquid column filled with culture medium.

[0123] The bioreactor 300 includes a first column 302 and a second column 304. In one of the embodiments shown in Figure 3, oxygen is introduced into the first column 302, while hydrogen or syngas is introduced into the second column 304; however, in other embodiments, their order may be reversed. Oxygen and / or hydrogen and / or syngas can be introduced into each of these columns, for example, by sparging, bubbling, and / or diffusing to move upward through the culture medium. The bioreactor 300 may include a horizontal liquid connection 312 at the top of the column and a horizontal liquid connection 314 at the bottom of the column.

[0124] In some embodiments, the level of the liquid medium in column 302 is maintained such that a gas headspace 316 is formed above the liquid. In addition, in some cases, the level of the liquid medium in column 304 can be positioned such that a gas headspace 318 is formed above the liquid medium. In some embodiments, the headspaces 316 and / or headspaces 318 can occupy at least about 2%, at least about 10%, at least about 25%, 2% to about 80%, about 10% to about 80%, or about 25% to about 80% of the total volume of the column in which they are located. The headspaces 316 and 318 can be isolated from each other by the liquid medium. In some embodiments, since the solubility of the gas in the liquid medium is low, it is possible to collect the gas at the top of the column after bubbling or diffusing the gas upward through each of those columns. By establishing isolated headspaces, it is possible to prevent dangerous amounts of hydrogen and oxygen gases from mixing with each other. For example, it is possible to prevent hydrogen gas in one column from mixing with oxygen gas in the other column (and vice versa). Preventing the mixing of hydrogen and oxygen gases can be achieved, for example, by maintaining the connection between the two columns to prevent the transport of gas from one column to the other by filling it with liquid. In some embodiments, the liquid culture medium is circulated between the first and second columns so that the headspace 316 and / or 318 can be maintained substantially steadily at the top of each of those columns.

[0125] In Figure 3, the horizontal liquid connections 312 at the top of the column and 314 at the bottom of the column are arranged such that, with the horizontal liquid connections constantly filled, the liquid medium flows upward through one column toward the oxygen gas and downward through the other column toward the hydrogen gas and / or syn gas. In other embodiments, the liquid medium can flow upward through a column containing hydrogen gas and / or syn gas and downward (countercurrently toward the gas) through the other column containing oxygen gas.

[0126] In some embodiments, the gas on one side or the other (but not both sides simultaneously) can be forcibly bubbling so that a particular column acts as an airlift reactor and drives the circulation of the culture medium between the two columns. In some embodiments, the circulation of the fluid can also be assisted by an impeller, turbine, and / or pump.

[0127] In some such embodiments, any unused hydrogen gas and / or syn gas (which may terminate at the headspace) that has passed through the culture medium without being taken up by microorganisms can be recirculated by pumping the gas out of the headspace, optionally compressing it, and returning it to the medium at the bottom of the liquid column on the hydrogen and / or syn gas sides. In some embodiments, oxygen and / or air can be similarly recirculated on their respective sides, or instead vented after passing through the headspace.

[0128] In certain embodiments, the oxyhydrogen microorganisms are freely circulated with the liquid medium between the first and second columns. In other embodiments, for example, the oxyhydrogen microorganisms are confined to the hydrogen side using a microfilter that allows the liquid medium to pass through but retains the microorganisms on the hydrogen side.

[0129] Figure 4 includes an illustrative schematic diagram of other operating methods of the bioreactor 300 that can be used in a particular embodiment. The bioreactor configuration in Figure 4 can take advantage of the relatively high solubility of carbon dioxide and / or the high ability of oxyhydrogen microorganisms to recover carbon dioxide from a relatively dilute stream. The operation illustrated in Figure 4 can take advantage of the carbon concentration mechanism inherent to oxyhydrogen microorganisms. Flue gas and / or air containing carbon dioxide can be transported through the oxygen side of the bioreactor. Carbon dioxide can be dissolved in solution and / or taken up by oxyhydrogen microorganisms and then transported to the hydrogen side of the reactor, for example, through the horizontal liquid connection 312 at the top of the column. On the hydrogen side, it is possible to provide a reducing equivalent that drives carbon fixation. In some embodiments, other gases (e.g., oxygen, nitrogen, etc.) pumped into the oxygen side have lower solubility compared to CO2 and are not brought into the hydrogen side. Rather than being sent from column 302 to column 304, the low-solubility gases can be transported into the headspace 316. In some embodiments, it is possible to vent the gas after it has been transported to the headspace 316.

[0130] Figure 5 includes an exemplary schematic diagram of an electrolytic apparatus 500 that can be used in a particular embodiment. The electrolytic apparatus 500 can be used, for example, as the unit exemplified as box 3 in Figure 1 labeled “Electron Donor Generation” and / or as the unit exemplified as box 3 in Figure 2 labeled “Electrolysis.” The electrolytic apparatus 500 can be designed to take advantage of the tolerance and need of oxyhydrogen microorganisms to specific concentrations of oxygen by reducing or eliminating the complete separation of hydrogen and oxygen produced from the electrolytic process, in contrast to the separation schemes used in conventional electrolytic systems designed for the production of pure hydrogen. The apparatus 500 includes an electrolytic unit 502 configured to produce H2 and O2 from water. Any suitable electrolytic unit 502 can be used to perform the electrolytic process. In some embodiments, it is possible to reduce the electrical resistance in the electrolytic unit 502 at the expense of complete separation of hydrogen and oxygen by means of one or more (but not limited to) of the following: namely, removing the separator used to prevent gas crossover in a standard electrolytic device and / or using a relatively short distance between the positive and negative electrodes.

[0131] The apparatus 500 may include an outlet 504 capable of transporting hydrogen and oxygen produced by the electrolysis unit 502. The outlet 504 may include a separator 506 that can be used to separate at least a portion of hydrogen from at least a portion of oxygen. In certain embodiments, a semipermeable membrane, such as a polymer membrane designed for H2 separation, can be used as the separator 506. In certain embodiments, the separator 506 may include metal foils, including (but not limited to) foils made of palladium, palladium alloys, vanadium, niobium, tantalum, and their alloys, and / or other metals and / or alloys that are permeable to hydrogen but not so permeable to other gases such as oxygen. In some embodiments, it is possible to separate hydrogen from oxygen using the separator so that the hydrogen content of one gas product exiting the separator is enriched to a level desirable for oxyhydrogen microorganisms. The gas product can then be transported to a bioreactor and used as a feedstock. In certain embodiments, the amount of hydrogen in one of the gas products exiting the separator can be set to a level that maximizes oxyhydrogen microbial activity and minimizes the loss of hydrogen generated via the electrolytic device 500.

[0132] The following documents are incorporated herein by reference in their entirety for all purposes: U.S. Provisional Patent Application No. 61 / 328,184, filed April 27, 2010, entitled "Use of Oxyhydrogen Microorganisms for the Recovery and Conversion of Non-Photosynthetic Carbon from Inorganic Carbon Sources to Useful Organic Compounds"; International Application PCT / US2010 / 001402, filed May 12, 2010, entitled "Biochemical Process Using Chemoautotrophic Microorganisms for Chemosynthetic Immobilization of Carbon Dioxide and / or Other Inorganic Carbon Sources to Organic Compounds and the Production of Other Useful Products"; and U.S. Patent Application Publication No. 2010 / 0120104, filed November 6, 2009, entitled "Biochemical Process Using Chemoautotrophic Microorganisms for Chemosynthetic Immobilization of Carbon Dioxide and / or Other Inorganic Carbon Sources to Organic Compounds and the Production of Other Useful Products". [Examples]

[0133] The following examples are intended to illustrate specific embodiments of the present invention and not to illustrate the entire scope of the invention.

[0134] Example 1 In this embodiment, oxyhydrogen microorganisms that accumulate high lipid content and / or other valuable compounds such as polyhydroxybutyrate (PHB) were grown in an inorganic medium containing CO2 as a carbon source and hydrogen acting as an electron donor, with oxygen providing the electron acceptor. Such oxyhydrogen microorganisms can be used in certain embodiments of the present invention for the conversion of C1 chemicals such as carbon dioxide into long-chain organic chemicals.

[0135] Cupriavidus necator DSM531 (which can accumulate high percentage cell aggregates as PHB) was inoculated into static anaerobic reactors. Inoculum was obtained from DSM medium no. 1 agar plates maintained under aerobic conditions at 28 degrees Celsius. Each anaerobic reactor contained 10 ml of liquid medium DSM no. 81 with 80% H2, 10% CO2, and 10% O2 in the headspace. The cultures were incubated at 28 degrees Celsius. After 8 days, Cupriavidus necator had an optical density (OD) of 0.98 at 600 nm and 4.7 × 10⁶ cells. 8 The cell density reached cells / ml.

[0136] Other propagation experiments were conducted with Cupriavidus necator (DSM531). The culture medium used for propagation was mineral salt medium (MSM) formulated by Schlegel et al. MSM medium was prepared by mixing 1000 ml of medium A, 10 ml of medium B, and 10 ml of medium C. Medium A contained 9 g / l Na2HPO4.12H2O, 1.5 g / l KH2PO4, 1.0 g / l, 0.2 g / l MgSO4.7H2O, and 1.0 ml of trace element medium. The trace element medium contained 1000 ml of distilled water, 100 mg / l ZnSO4.7H2O, 30 mg / l MnCl2.4 H2O, 300 mg / l H3BO3, 200 mg / l COCl2.6H2O, 10 mg / l CuCl2.2H2O, 20 mg / l NiCl2.6H2O, and 30 mg / l Na2MoO4.2H2O. Medium B contained 100 ml of distilled water, 50 mg ferric ammonium citrate, and 100 mg CaCl2. Medium C contained 100 ml of distilled water and 5 g NaHCO3.

[0137] Cultures were grown in 20 ml of MSM medium in a 150 ml sealed serum vial with the following gas mixture in headspace: 71% hydrogen, 4% oxygen, 16% nitrogen, and 9% carbon dioxide. The headspace pressure was 7 psi. Cultures were grown at 30 degrees Celsius for 8 days. Cupriavidus necator reached an OD of 0.86 at 600 nm.

[0138] It is well known that larger-scale bioreactor systems can achieve faster growth rates and higher cell densities. Therefore, it is thought that higher growth rates and cell densities can be achieved simply by scaling up the systems described above. For example, Cupriavidus necator, also known as Alcaligenes eutrophus, Ralstonia eutropha, and Hydrogenomona eutropha, was grown in an H2 / CO2 / O2-based bioreactor to a cell density of over 90 grams / liter [Tanaka, Ishizaki; Biotech. And Bioeng., vol.45, 268-275 (1995)] with a doubling time of less than 2 hours [Ammann, Reed, Durichek, Appl. Microbio., (1968) 822-826].

[0139] This specification has described in sufficient detail certain preferred embodiments of the invention so that those skilled in the art can practice the full scope of the invention. However, many possible variations of the invention that have not been specifically described are still considered to be included within the scope of the invention and the appended claims. Thus, these descriptions given herein are merely illustrative and are not intended to limit the scope of the invention in any way. More generally, those skilled in the art will readily understand that all parameters, dimensions, materials, and configurations described herein are illustrative, and that the actual parameters, dimensions, materials, and / or configurations will depend on one or more specific applications in which the teachings of the invention are used. Those skilled in the art will be able to identify and verify many equivalent forms to the specific embodiments of the invention described herein without going beyond ordinary experimentation. Thus, since the above embodiments are provided merely as examples, it should be understood that the invention can be practiced in forms other than those specifically described and claimed, within the scope of the appended claims and their equivalents. The invention relates to each of the individual features, systems, articles, materials, kits, and / or methods described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods is included within the scope of the present invention, provided that such features, systems, articles, materials, kits, and / or methods are not contradictory to each other.

[0140] In this specification and in the claims, the indefinite articles "a" and "an" shall be deemed to mean "at least one" unless otherwise clearly specified.

[0141] As used herein and in the claims, the expression “and / or” shall be deemed to mean “one or both” of the elements thus combined, that is, elements that exist in some cases together and in other cases separately. Where appropriate, other elements not specifically identified by the “and / or” phrase may exist, whether related to the specifically identified elements or not, unless otherwise clearly specified. Thus, for example, the reference term “A and / or B” when used in combination with open-ended language such as “comprising,” may mean in one embodiment A but not B (including elements other than B), in another embodiment B but not A (including elements other than A), and in yet another embodiment both A and B (including other elements).

[0142] As used herein and in the claims, “or” shall have the same meaning as “and / or” as defined above. For example, when separating enumerated items, “or” or “and / or” shall be inclusive; that is, it shall include not only several elements or at least one of the enumerated elements, but more than one, and optionally additional items not enumerated. Only terms that clearly indicate otherwise, such as “only one of” or “exactly one of” or, as used in the claims, “consisting of,” shall mean that it includes several elements or exactly one of the enumerated elements. Generally, as used herein, the term “or” shall be considered only an exclusive choice (i.e., “one or the other, but not both”) when preceded by an exclusive term such as “either,” “one of,” “only one of,” or “exactly one of.” As used in the claims, "consisting essentially of" shall have its usual meaning as it is used in the field of patent law.

[0143] In the claims, as in the above specification, all transitional phrases such as “including,” “carrying,” “having,” “containing,” “involving,” and “holding” shall be open-ended; that is, they shall mean inclusion without limitation. Only the transitional phrases “more than” and “more essentially than” shall be closed or semi-closed transitional phrases, respectively.

Claims

1. A biochemical method for the recovery and conversion of inorganic carbon compounds and / or organic compounds containing only one carbon atom into organic chemicals, Introducing inorganic carbon compounds and / or organic compounds containing only one carbon atom into an environment suitable for the retention of oxyhydrogen microorganisms and / or capable of retaining extracts of oxyhydrogen microorganisms, Within the aforementioned environment, the inorganic carbon compound and / or the organic compound containing only one carbon atom are converted into the organic chemical and / or its precursor via at least one chemically synthesized carbon fixation reaction using a cell extract containing the oxyhydrogen microorganism and / or an enzyme derived from the oxyhydrogen microorganism. Includes, The method wherein the chemical synthesis immobilization reaction is driven at least in part by chemical and / or electrochemical energy provided by electron donors and electron acceptors that are chemically and / or electrochemically generated and / or introduced into the environment from at least one external source outside the environment.

2. The method according to claim 1, wherein the inorganic carbon compound contains carbon dioxide.

3. The method according to claim 1 or 2, wherein carbon dioxide is contained by dissolving carbon dioxide gas alone and / or in a mixture or solution further containing carbonate ions and / or bicarbonate ions.

4. The method according to any one of claims 1 to 3, wherein the inorganic carbon includes inorganic carbon contained in a solid phase.

5. The method according to any one of claims 1 to 4, wherein the organic compound containing only one carbon atom comprises carbon monoxide, methane, methanol, formate, and / or formic acid.

6. The method according to any one of claims 1 to 5, wherein an organic compound containing an electron donor and / or a single carbon atom is produced by gasification and / or thermal decomposition of an organic substance and provided to an oxyhydrogen microorganism as syngas.

7. The method according to any one of claims 1 to 5, wherein an organic compound containing an electron donor and / or a single carbon atom is produced via methane steam reforming and provided to an oxyhydrogen microorganism as syngas.

8. The method according to claim 6 or 7, wherein the hydrogen-to-carbon monoxide ratio in the syngas is adjusted via a water-gas shift reaction before the syngas is delivered to oxyhydrogen microorganisms.

9. The method according to any one of claims 1 to 8, wherein the oxyhydrogen microorganisms include oxyhydrogen microorganisms selected from one or more of the following categories: purple non-sulfur photosynthetic bacteria, cyanobacteria, and / or green algae.

10. The method according to any one of claims 1 to 9, wherein the oxyhydrogen microorganisms include oxyhydrogen microorganisms selected from cyanobacteria and / or green algae.

11. Acid-hydrogen microorganisms include species of the following genera: Rhodopseudomonas, Rhodospirillum, Rhodococcus, Rhizobium, Thiocapsa, and Pseudomonas. Species of the genus Hydrogenomonas, Hydrogenobacter, Hydrogenovibrio, Helicobacter, Xanthobacter, and Hydrogenophaga. The method according to any one of claims 1 to 9, comprising an acid-hydrogen microorganism selected from one or more of the following: species of Hydrogenophaga, species of Bradyrhizobium, species of Ralstonia, species of Alcaligenes, species of Variovorax, species of Acidovorax, species of Anabaena, species of Scenedesmus, species of Chlamydomonas, species of Ankistrodesmus, and species of Rhaphidium.

12. Oxyhydrogen microorganisms include species of the following genera: Rhodospirillum, Rhizobium, Thiocapsa, Hydrogenovibrio, Helicobacter, Xanthobacter, Hydrogenophaga, and Bradyrhizobium. The method according to any one of claims 1 to 9, comprising an acid-hydrogen microorganism selected from one or more of the following: species of the genus Variovorax, species of Acidovorax, species of Anabaena, species of Scenedesmus, species of Chlamydomonas, species of Ankistrodesmus, and species of Rhaphidium.

13. The electron donor is a reducing agent, namely ammonia, ammonium, carbon monoxide, dithionite, elemental sulfur, hydrocarbons, hydrogen, metabisulfite, nitrous oxide, nitrite, for example, sodium thiosulfate (Na ２ S ２ O ３ ) or calcium thiosulfate (CaS ２ O ３ The method according to any one of claims 1 to 12, comprising, but not limited to, sulfates such as thiosulfates, sulfides such as hydrogen sulfide, sulfites, thionates, thionic acid, transition metals or their sulfides, oxides, chalcogenides, halides, hydroxides, oxyhydroxides, phosphates, sulfates, or carbonates in the dissolved phase or solid phase, and one or more of conduction band electrons or valence band electrons in a solid electrode material.

14. The method according to any one of claims 1 to 13, wherein the electron acceptor comprises one or more of the following: carbon dioxide, oxygen, nitrite, nitrate, ferric ion or other transition metal ion, sulfate, or valence band holes or conduction band holes in a solid electrode material.

15. The method according to any one of claims 1 to 14, wherein one or more chemical pretreatment steps are performed prior to the conversion step, wherein electron donors and / or electron acceptors are generated and / or purified from at least one input chemical, and / or recycled from chemicals generated during the fixation step and / or from a waste stream from other industrial processes, mining processes, agricultural processes, sewage processes, or waste generation processes.

16. The method according to any one of claims 1 to 15, wherein, after the conversion step, one or more process steps are performed to separate the chemically synthesized organic and / or inorganic chemical products from the process stream generated during the conversion step and to process them to produce products in a form suitable for storage, transport, and sale, and further, one or more process steps are performed to separate the cell aggregates from the process stream and recirculate them into the environment, and / or collect them and to process them to produce biomass in a form suitable for storage, transport, and sale.

17. The method according to any one of claims 1 to 16, wherein one or more process steps are performed after the conversion step to remove waste products and / or impurities and / or contaminants from the process stream generated during the stagnation step and dispose of them.

18. The method according to claim 17, wherein the waste product includes waste product from a nutrient medium used to maintain the oxyhydrogen reaction.

19. The method according to any one of claims 1 to 18, wherein one or more process steps are performed after the conversion step, to recirculate any remaining unused nutrients and / or process water after removing the oxyhydrogen cell aggregates and / or chemical by-products and / or waste products or contaminants of the process stream generated during the immobilization step, and return them to the environment to support further chemosynthesis.

20. The method according to any one of claims 1 to 19, wherein electron donors and / or electron acceptors are generated or recycled using a renewable power source, an alternative power source, or a conventional power source with low greenhouse gas emissions, and the power source is selected from at least one of photovoltaic, solar thermal, wind, hydroelectric, nuclear, geothermal, enhanced geothermal, ocean thermal, wave power, and tidal power.

21. The electron donor is one of the following: elemental Fe ０ , siderite (FeCO ３ ), magnetite (Fe ３ O ４ ), pyrite or marcasite (FeS ２ ), pyrrhotite (Fe （１－ｘ） S (x = 0 to 0.2)), pentlandite (Fe,Ni) ９ S ８ , violarite (Ni ２ FeS ４ ), nickeline (Ni,Fe)S ２ , arsenopyrite (FeAsS), or other iron sulfides, realgar (AsS, orpiment (As ２ S ３ ), cobaltite (CoAsS), rhodochrosite (MnCO ３ ), chalcopyrite (CuFeS ２ ), bornite (Cu ５ FeS ４ ), covellite (CuS), tetrahedrite (Cu ８ Sb ２ [[ID=3৮]]S ７ ), enargite (Cu ３ AsS ４ ), tennantite (Cu １２ As ４． S １３ ), chalcocite (Cu ２ S), or other copper sulfides, sphalerite (ZnS), marmatite (ZnS), or other zinc sulfides, galena (PbS), jamesonite (Pb ５ (Sb,As ２ )S ８ ), or other lead sulfides, argentite or acanthite (Ag ２ S), molybdenite (MoS ２ ), millerite (NiS), polydymite (Ni ３ S ４ ), or other nickel sulfides, stibnite (Sb ２ S [[ID=६9]] ３ ), Ga ２ S ３ , CuSe, cooperite (PtS), laurite (RuS ２ ), braggite (Pt, Pd, Ni)S, FeCl ２ The method according to any one of claims 1 to 20, wherein the product is generated from a naturally occurring mineral selected from one or more of the following.

22. The electron donor is one of the following: process gas, tail gas, fortified petroleum recovery vent gas, biogas, acidic mine wastewater, landfill leachate, landfill gas, geothermal gas, geothermal sludge or brine, metallic contaminants, gangue, tailings, sulfides, disulfides, mercaptans selected from one or more of methyl mercaptan, dimethyl mercaptan, and ethyl mercaptan, carbonyl sulfide, carbon disulfide, alkanesulfonates, dialkyl sulfides, thiosulfates, thiofurans, thiocyanates, isothiocyanates. The method according to any one of claims 1 to 21, which is produced from a contaminant or waste product selected from one or more of the following: anates, thioureas, thiols, thiophenols, thioethers, thiophenes, dibenzothiophenes, tetrathionates, dithionites, thions, dialkyl disulfides, sulfones, sulfoxides, sulfolanes, sulfonic acids, dimethyl sulfoniopropionates, sulfonic acid esters, hydrogen sulfide, sulfate esters, organosulfurs, sulfur dioxide, and all other sour gases.

23. The delivery of the reduced equivalent from an electron donor to an acid-hydrogen microorganism for one or more chemosynthetic reactions during the immobilization process is facilitated by introducing a hydrogen storage material into the environment in the form of a solid carrier medium for microbial growth, which allows the absorbed or adsorbed hydrogen electron donor to be placed in close proximity to the chemoautotrophic organism. ２ The method according to any one of claims 1 to 22, which is kinetically and / or thermodynamically enhanced by one or more of the following: introducing an electron carrier that helps transfer reducing power from an electron-poorly soluble electron donor containing electrons in a gas or solid electrode material to a chemoautotrophic culture medium; and directly introducing an electrode material in the form of a solid growth carrier medium into the environment that facilitates placing solid electrons in close proximity to chemoautotrophic organisms.

24. The method according to any one of claims 1 to 23, wherein the organic chemical comprises a compound having a carbon chain length of C5 to C30.

25. The method according to any one of claims 1 to 24, wherein at least one chemosynthetic reaction is carried out by an improved, optimized, or engineered oxyhydrogen microorganism to fix inorganic carbon compounds and / or organic compounds containing only one carbon atom, via a method comprising one or more of the following: accelerated mutagenesis, genetic engineering or genetic modification, hybridization, synthetic biology, and traditional selective breeding, the immobilization of inorganic carbon compounds and / or organic compounds containing only one carbon atom, and the production of organic compounds.

26. The first tower, which includes the upper and lower parts, The second tower, which includes the upper and lower parts, A bioreactor comprising, wherein the upper portion of the second tower is fluidly connected to the upper portion of the first tower, and the lower portion of the second tower is fluidly connected to the lower portion of the first tower, When circulating the liquid between the first and second towers, the gas volume is constructed and arranged such that it becomes substantially constant at the top of the first and / or second tower, A bioreactor in which the volume of gas occupies at least approximately 2% of the total volume of the tower in which it is located.

27. The bioreactor according to claim 26, wherein the bioreactor contains a liquid culture medium.

28. The bioreactor according to claim 26, wherein the bioreactor contains oxyhydrogen microorganisms.

29. A method for operating a bioreactor, comprising circulating a liquid containing a growth medium between a first column and a second column, wherein during the operation, the volume of gas is maintained substantially steadily at the top of the first column and / or the second column, and the volume of gas accounts for at least about 2% of the total volume of the column in which it is located.

30. The method according to claim 29, wherein the volume of the gas includes hydrogen and / or oxygen.

31. The method according to claim 29, wherein the volume of the gas contains hydrogen.

32. The method according to claim 29, wherein the volume of the gas accounts for approximately 2% to approximately 10% of the total volume of the tower in which it is located.

33. The method according to claim 29, wherein the first and / or second towers include an inlet and at least a portion of the gas located at the top of the towers is recirculated from the volume through the inlet of the first and / or second towers and returned to the first and / or second towers.

34. The method according to claim 29, wherein the bioreactor contains oxyhydrogen microorganisms.

35. A chamber constructed and arranged to electrolyze water to produce oxygen and hydrogen, An outlet section including a separator constructed and positioned to separate at least a portion of the oxygen in the stream from at least a portion of the hydrogen in the stream, so that the hydrogen content of the fluid exiting the separator is suitable for use as a feed stream to a reactor containing a culture of oxyhydrogen microorganisms, An electrolytic device, including one.

36. The electrolytic apparatus according to claim 35, wherein the outlet of the electrolytic apparatus is connected to a bioreactor in a fluid communication state.

37. The electrolytic apparatus according to claim 36, wherein the bioreactor is constructed and arranged for the recovery and conversion of inorganic carbon compounds and / or organic compounds containing only one carbon atom into organic chemicals by oxyhydrogen microorganisms.

38. A method for operating an electrolytic apparatus, comprising electrolyzing water to produce a first stream containing oxygen and hydrogen; and This includes separating at least a portion of the oxygen from at least a portion of the hydrogen to produce a second stream that is relatively richer in hydrogen compared to the first stream. The method wherein the second stream is suitable as a supply stream to a reactor containing a culture of oxyhydrogen microorganisms.

39. The method according to claim 38, comprising supplying a second stream to a bioreactor.

40. The method according to claim 39, wherein the bioreactor contains oxyhydrogen microorganisms.

Images