Processes for the biotechnological production of bioproducts

Abstract

The present invention relates to a process and an apparatus for the biotechnological production of a bioproduct. The process comprises a first and a second bioprocess carried out in a first and a second bioreactor (2, 3) and a tank (4). The first bioprocess is a carbon dioxide producing bioprocess resulting in the production of a bioproduct, and the second bioprocess is a chemoautotrophic bioprocess in which the CO2 produced in the first bioprocess is consumed by hydrogen bacteria oxidizing H2 with O2 as the electron acceptor. The electrolytically produced H2 and O2 are separated and O2 is fed directly to the second and / or first bioprocess, while H2 is initially dissolved in a medium contained in the tank (4). The H2 saturated medium is then fed to the second bioprocess. The biomass produced in the second bioprocess is fed to the first bioprocess as an additional C and N source.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a process for the biotechnological production of bioproducts. Background Art

[0002] The biotechnological production of bioproducts, for example biobased chemicals, has long been known, and it is expected to provide chemicals in a method with improved sustainability and environmental friendliness for commercial use in, for example, the chemical industry. By way of example, so-called biofuels such as bioethanol or biodiesel are produced in bioprocesses to replace fossil fuels derived from petroleum (see, for example, Patent Document 1 and Patent Document 2). However, in view of global warming caused by the increasing amount of carbon dioxide (CO₂) in the atmosphere, bioprocesses for large-scale production of chemicals must also minimize CO₂ emissions to be sustainable.

[0003] Both Patent Document 1 and Patent Document 2 describe fixation of CO₂ produced in a first bioprocess in a second bioprocess for reusing the produced CO₂. Patent Document 1 discloses a method for converting a carbon source into lipids, wherein lipid production is carried out in an aerobic fermenter using the oleaginous yeast *Yarrowia lypolytica*, and carbon dioxide generated during lipid production is converted into a carbon substrate by CO₂ fixation carried out in an anaerobic fermenter by bacteria of the genus *Clostridium sp.* These Clostridium bacteria use H₂ gas produced by electrolysis of water and supplied into the anaerobic fermenter as a reducing agent. Patent Document 2 discloses production of biofuel using photosynthetic plant cells that fix produced CO₂, for example by yeast cells that produce biofuel.

[0004] Patent document 3 describes, in particular, a method for the collection and fixation of oxyhydrogen ("Knallgas") carbon dioxide from flue gas, for example, by Knallgas bacteria, in which oxygen and hydrogen can be provided, for example, by the electrolysis of water. The biotechnological production of biofuels using Knallgas bacteria is also described in non-patent document 1. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] International Publication No. 2011 / 088364(A9) [Patent Document 2] International Publication No. 2009 / 133351(A2) [Patent Document 3] International Publication No. 2011 / 139804(A2) [Non-patent literature]

[0006] [Non-Patent Document 1] Brigham, C., "Perspectives for the biotechnological Production of biofuels from CO2 and H2 using Ralstonia eutropha and other 'Knallgas' bacteria," Applied Microbiology and Biotechnology, Vol. 103 (2019), 10.1007 / s00253-019-09636-y) [Overview of the project] [Problems that the invention aims to solve]

[0007] The objective of this invention is to improve the biotechnological production of bioproducts, particularly biochemicals, with respect to sustainability, especially minimizing net carbon dioxide production. [Means for solving the problem]

[0008] In a first aspect, the present invention relates to a process for the biotechnological production of bioproducts, a) A first bioprocess comprising culturing a first type of microorganism in a first medium at a first pressure p1 in a first bioreactor, the first bioprocess resulting in the production of a bioproduct and carbon dioxide CO2, b) A second bioprocess comprising culturing a second type of microorganism in a second medium at a second pressure p2 in a second bioreactor, the second bioprocess being a process that produces biomass and consumes molecular oxygen O2, molecular hydrogen H2, and at least a portion of the CO2 produced in the first bioprocess of step a), c) The step of electrolyzing water into O2 and H2, supplying at least a portion of the O2 produced by electrolysis to a second bioprocess and / or a first bioprocess, and supplying at least a portion of the H2 produced by electrolysis to a headspace and a tank containing a third medium at a third pressure p3, wherein the third medium is identical to the second medium in the second bioreactor. d) A step of supplying at least a portion of a third medium containing H2 from the tank to the second bioprocess, The process includes a process in which at least a portion of the biomass produced in the second bioprocess of step b) is used as a C source for the first bioprocess of step a), and p3 ≥ p2 ≥ p1.

[0009] This process combines two bioprocesses, the first being a CO2 production process and the second being a CO2 consumption bioprocess, which includes non-photosynthetic biofixation of carbon dioxide produced in the first bioprocess by microorganisms that oxidize hydrogen and reduce oxygen, such as "Knurl gas" bacteria. The first and second bioprocesses are run spatially separated from each other in separate bioreactors. The CO2 produced in the first bioprocess is supplied to the second bioprocess to be consumed at least partially, preferably at least almost entirely, or completely in the second bioprocess. The process of the present invention is preferably a "CO2 neural" process, i.e., a process in which the produced carbon dioxide is consumed at least essentially completely, and therefore no CO2 is released into the environment. The biomass produced in the second bioprocess is used as an additional carbon source for the first bioprocess, along with at least some of the CO2 from the first bioprocess. The biomass may also be used as an additional nitrogen source. Furthermore, oxygen gas, produced by the electrolysis of water and used as an electron acceptor by microorganisms in the second bioprocess, is supplied directly to the second bioprocess, or even further, to the first bioprocess. Hydrogen gas, produced by electrolysis and used as an electron donor by microorganisms in the second bioprocess, is supplied indirectly to the second bioprocess, not directly, but through a medium contained in a separate tank. The advantage of this separate supply of oxygen and hydrogen to the second bioprocess is, for example, increased safety with respect to explosion protection. Moreover, this serves as a countermeasure against the poor solubility of H2 in the aqueous medium.

[0010] The term "bioplastic product" refers to compounds, particularly organic compounds, that are components of chemical reactions in living cells, produced by chemical reactions in living cells, or involved in chemical reactions in living cells, in contrast to products of bioprocesses, such as chemicals based on fossil fuels. Biochemicals derived from biomass may be structurally identical or structurally different from existing chemicals derived from fossil fuels. Examples of biochemicals include, for example, fatty acids and fatty acid derivatives, such as lipids, alcohols, such as diols, and acids, biopolymers or monomers for polymerization, peptides, etc. This term also encompasses biomass as products of bioprocesses, such as living, freeze-dried, or hydrolyzed microorganisms. As defined above, the term "bioplastic product" refers to compounds, particularly organic compounds, that are components of chemical reactions in living cells, produced by chemical reactions in living cells, or involved in chemical reactions in living cells. The term "biochemical" is sometimes used synonymously with "bioplastic product." While this invention may be described, and is primarily intended, in relation to the production of biochemical substances, it should be noted that this should not be interpreted as excluding, for example, the production of biomass as a bioproduct.

[0011] The term "lipid" refers to organic compounds that are soluble in nonpolar organic solvents (such as chloroform or ether) and are normally insoluble in polar solvents such as water, and includes, for example, fats and oils, oils, waxes, phospholipids, and steroids.

[0012] As used herein, the term “bioprocess” refers to any biological process, i.e., a process involving living cells, particularly living microorganisms such as bacteria, or functional components of living cells, such as enzymes, cell-free systems, or organelles, in order to obtain a desired product. The product may be a compound, such as an organic compound, a mixture of compounds, such as a mixture of organic compounds, or biomass. An example is the production of lipids from substrates such as glucose, starch, glycerol, or analogues.

[0013] The term "bioreactor" is understood to mean a reactor containing a working volume that can provide a suitable medium, such as an aqueous medium suitable for supporting the growth of a culture of microorganisms, such as bacterial cells, or that can carry out biological and / or biochemical processes and reactions, or that can create conditions for carrying out biological and / or biochemical processes and reactions. These processes and reactions may be material transformations, such as the synthesis, modification, or degradation of substances by living or cell-free systems, or biomass growth, i.e., the growth of living cells, such as bacterial cells. Depending on the type of process to be carried out and / or the type of microorganism used to carry out the process, the bioreactor may be equipped with, for example, equipment for aeration / ventilation and / or stirring of the medium.

[0014] The term “closed” in reference to a bioreactor or tank means that the photobioreactor or tank is enclosed from the surrounding environment so that it can be pressurized. This does not exclude the possibility that the bioreactor or tank has openings, connections or similar features that allow fluids, gases, CO2, or liquids to be introduced into or removed from the container.

[0015] As used herein, the terms “Knurl gas bacteria,” “hydrogen bacteria,” “H2 bacteria,” or “acid-hydrogen bacteria” refer to a physiologically defined group of bacteria that grow autotrophically, that is, that they can oxidize hydrogen (H2) and fix carbon dioxide while using oxygen (O2) as the final electron acceptor. The abbreviation HOB may be used instead of the terms hydrogen bacteria or H2 bacteria. The term “acid-hydrogen” may be used synonymously with “Knurl gas.” “Knurl gas” is a mixture of gaseous hydrogen and gaseous oxygen. Knurl gas bacteria are aerobic, arbitrarily chemosynthetic inorganic autotrophic bacteria. Examples of Kunar gas bacteria include Hydrogenophilus thermoluteolus, Hydrogenobacter thermophilus, Hydrogenovibrio marinus, Cupriavidus metallidurans (formerly Alcaligenes eutropha or Ralstonia eutropha), Rhodococcus opacus, Xantobacter autotrophicus, and Cupriavidus necator. The following simplified general reaction scheme describes aerobic CO2 fixation by Kunar gas bacteria via H2 oxidation using O2 as an electron acceptor. H2 + O2 + CO2 → Biomass + H2O

[0016] The term "chemotrophic" refers to organisms that use the oxidation of compounds as an energy source, in contrast to "phototrophic" organisms that derive their own energy from light. The term "chemolithotrophic" refers to organisms that use inorganic compounds, such as hydrogen gas, as electron donors, in contrast to the term "chemoorganotrophic," which refers to organisms that use organic compounds as electron donors. The terms "autotroph" or "heterotroph" refer to organisms whose carbon source is CO2 in "autotrophic" organisms and organic carbon compounds in "heterotrophic" organisms. The term "mixotrophic" is sometimes used for organisms that can use both organic compounds and CO2 as carbon sources. The term "chemolithoautotrophic" refers to organisms that use inorganic compounds as an energy source and electron donor, and CO2 as a carbon source. The term "facultative chemolithoautotrophic" refers to organisms that can also grow chemoorganoheterotrophically. For example, the "Knurl gas" bacteria grow heterotrophically when H2 is at tropospheric concentration, and chemosynthetic inorganically only when H2 is available at higher concentrations (Pumphrey GM, Ranchou-Peyruse A, Spain, JC., "Cultivation-independent detection of autotrophic hydrogen-oxidizing bacteria by DNA stable-isotope probing," Applied Environmental Microbiology, vol. 77, no. 14, pp. 4931-4938 (2011), doi:10.1128 / AEM.00285-11).

[0017] The term "yeast" refers to eukaryotic single-celled microorganisms. Examples of yeast include Saccharomyces cerevisiae, Candida albicans, and Yarowia liporitica. The term "oleaginous yeast" refers to yeast that can store more than 20% of its cell dry weight in the form of lipids (e.g., fats), e.g. Abeln, F., Chuck, CJ, "The history, state of the art and future prospects for oleaginous yeast research," Microbial Cell Factories, Vol. 20, p. 221 (2021), doi:10.1186 / s12934-021-01712-1; Blomqvist, J., Pickova, J., Tilami, SK et al., "Oleaginous yeast as a component in fish feed," Scientific Reports. (See Reports), Vol. 8, p. 15945 (2018), doi:10.1038 / s41598-018-34232-x). Examples of oily yeasts include Cutaneotrichosporon oleaginosus, Rhodotorula toruloides, Yarowia liporitica, Lipomyces starkeyi, Trichosporon oleaginosus (formerly Cryptococcus curvatus), and Rhodosporidium toruloides.

[0018] The term "water electrolysis" relates to a process in which direct current is used to split water into hydrogen and oxygen. The term "electrolyzer" is used to refer to a device capable of electrolyzing water.

[0019] Unless explicitly stated or otherwise unambiguously clear from the context, the term "hydrogen" refers to dihydrogen, i.e., the elemental molecule H2 consisting of two hydrogen atoms bonded via a single covalent bond. In place of H2, the terms "molecular hydrogen" or "gaseous hydrogen" may also be used synonymously.

[0020] Unless explicitly stated or otherwise unambiguously clear from the context, the term "oxygen" refers to the elemental molecule O2 consisting of two oxygen atoms bonded via a single covalent bond. In place of O2, the terms "molecular oxygen" or "gaseous oxygen" may also be used synonymously.

[0021] The term "C source" or "carbon source" relates to a compound comprising at least one carbon atom that can be, or is, incorporated into compounds constituting cellular material. In particular, an example of a C source for autotrophic bacteria is CO2.

[0022] The phrase "at least a portion of the biomass produced in the second bioprocess is used as a C source for the first bioprocess" should not be construed to mean that said biomass is the sole carbon source for the first bioprocess. Rather, the biomass is used as an additional carbon source employed in the first bioprocess. Furthermore, this phrase does not exclude that said biomass may also be used for other purposes, for example as an additional nitrogen source.

[0023] The term "CO2 fixation" or "carbon fixation" relates to the biological incorporation of inorganic carbon, in particular in the form of carbon dioxide, into organic carbon compounds, such as carbohydrates. The term "non-photosynthetic biological fixation of carbon dioxide" relates to carbon fixation that does not involve photosynthesis, i.e., carbon fixation by non-photosynthetic organisms.

[0024] The term "fermentation" or "fermenting" refers to any process in which microbial activity brings about a desirable change in a compound, food, or beverage.

[0025] The expression "the third medium is identical to the second medium" means that the third medium and the second medium have at least the same composition, that is, the same components at essentially the same concentration. This term does not implicitly mean that the concentrations of the dissolved gases, such as H2, O2, and CO2, are identical.

[0026] The phrase "supplying at least a portion of the O2 produced by electrolysis to a second bioprocess and / or a first bioreactor" includes cases where all or part of the electrolyzed O2 is supplied to the first bioprocess only, or where all or part of the electrolyzed O2 is supplied to the first bioprocess only, or where at least a portion of the electrolyzed O2 is supplied to the first bioprocess and / or to the second bioprocess, simultaneously or intermittently. The terms "a portion" or "partially" may refer to a portion of a continuous total gas flow or a portion of a time-divided O2 gas flow. Thus, a portion of the continuous flow of oxygen leading to the second bioprocess may be separated from the total oxygen flow and used for oxygen supply to the first bioprocess (see below), and / or the total oxygen gas flow may be supplied, for example, to the second bioprocess during the first period and to the first bioprocess during the second period. The formulation that oxygen is supplied "directly" to the second bioprocess does not exclude the possibility that the oxygen is interimly stored in a storage tank, such as a gas cylinder.

[0027] The statement, "The carbon dioxide produced in the first bioprocess is recovered from the first bioprocess and supplied to the second bioprocess," should not be interpreted as meaning that CO2 is selectively recovered from the first bioprocess and supplied to the second bioprocess. Rather, it should be interpreted as meaning that CO2-containing air is recovered from the first bioprocess, for example, from the headspace of the first bioreactor, and CO2-enriched or concentrated CO2-containing air is optionally supplied to the second bioreactor.

[0028] In the process of the present invention, the electrolyzed oxygen may, in one embodiment, be supplied only to the second bioprocess for the oxygen supply of a second type of microorganism. In an alternative embodiment, the electrolyzed oxygen may be supplied only to the first bioprocess, which in this embodiment is preferably an aerobic bioprocess or at least a microaerophilic bioprocess. In this embodiment, the oxygen supply to the aerobic second bioprocess is provided by a gaseous stream containing CO2 produced in the first bioprocess, which is recovered from the first bioprocess and supplied to the second bioprocess. Thus, the second bioprocess is supplied with residual oxygen from the first bioprocess. In another embodiment, the electrolyzed oxygen is partially supplied to the first bioprocess and partially supplied to the second bioprocess. The oxygen may be divided in terms of quantity or time, i.e., divided into two partial streams that are supplied continuously to the first and second bioprocesses or intermittently to these bioprocesses.

[0029] In a preferred embodiment of the process of the present invention, the third pressure p3 is greater than or equal to the second pressure p2, and the second pressure p2 is greater than or equal to the first pressure p1. Particularly preferred, the third pressure p3 is greater than the second pressure p2, and the second pressure is greater than the first pressure p1, i.e., p3 > p2 > p1. Even more preferred, p1 is greater than atmospheric pressure. The greater the pressure p3 in the tank having the third medium to which H2 gas is supplied, the greater the amount of H2 that can be dissolved in the third medium due to the pressure dependence of the gas solubility. When the pressure p2 in the second bioreactor is lower than the pressure p3, the release of H2 into the second medium becomes easier. The tank may be configured as the third bioreactor.

[0030] The first bioprocess may be any bioprocess that results in the production of a desired bioproduct and carbon dioxide (CO2). Preferably, the first bioprocess is a bioprocess that does not involve the production of a strongly reducing gas such as H2 or CO. The first bioprocess may be an aerobic process, an anaerobic process, or a microaerophilic process. Preferably, the first bioprocess is an aerobic process or at least a microaerophilic process. The bioproduct may be recovered from the first bioprocess by any method appropriate to each bioproduct, for example, if the bioproduct is stored in cells, by recovering at least a portion of the cells cultured in the first bioprocess. The cells recovered from the first bioprocess may then be further processed to obtain the bioproduct. The first bioprocess may also be a bioprocess for the production of biomass, for example, a mass of living microorganisms grown in the first bioprocess. In a preferred embodiment of the process according to the present invention, the first bioprocess is an aerobic bioprocess. More preferably, the first bioprocess is a fermentation process for the production of lipids. Preferably, the fermentation process includes cultivating an oily yeast, preferably Rhodosporidia tolloides.Alternatively, the first bioprocess may involve the production of carotenoids, preferably using Rhodosporidia tolloides (e.g., Igreja, WS; Maia, FdA; Lopes, AS; Christe, RC, "Biotechnological Production of Carotenoids Using Low Cost-Substrates Is Influenced by Cultivation Parameters: A Review," International Journal of Molecular Sciences). Sciences), Vol. 22, p. 8819 (2021), doi:10.3390 / ijms22168819; Yaegashi, J., Kirby, J., Ito, M., et al., "Rhodosporidium toruloides: a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts," Biotechnology For Biofuels And Bioproducts, Vol. 10, p. 241 (2017), see doi:10.1186 / s13068-017-0927-5). The microorganisms used in the first bioprocess, such as yeast cells, may be genetically engineered.

[0031] In the case of a first aerobic bioprocess, the oxygen produced by electrolysis may be supplied only to the first bioprocess, or it may be partially supplied to the first bioprocess to support the aerobic process. The latter may be carried out, for example, by a) splitting the O2 gas flow into two gas flows, one supplied to the second bioprocess and the other to the first bioprocess, or b) supplying the entire O2 gas flow intermittently to the first or second bioprocess. A combination of both options a) and b) is also possible. As described above, it is also possible that oxygen is supplied only to the first bioprocess, and the second bioprocess is supplied with oxygen by a gas flow containing CO2 recovered from the first bioprocess.

[0032] In a preferred embodiment of the process of the present invention, the second bioprocess is a non-photosynthetic bioprocess and includes chemosynthetic inorganic autotrophic consumption of O2, H2, and CO2. In this embodiment, the second bioprocess is a bioprocess in which knar gas bacteria grow, oxidizing H2 with O2 as an electron acceptor and fixing carbon dioxide. Preferably, at least a large portion, more preferably all, of the carbon dioxide is produced from the first process. The microorganisms used in the second bioprocess may be genetically engineered.

[0033] A particularly preferred first bioprocess is an aerobic process for the fermentation production of lipids and / or carotenoids, comprising the culture of yeast cells, preferably of the genus Rhodosporidi, particularly preferably Rhodosporidi tolloides; and a second bioprocess comprises the chemosynthetic inorganic autotrophic culture of cnar gas bacteria, preferably of the genus Hydrogenophilus, particularly preferably Hydrogenophilus thermortheorus, with H2 and O2. O2 produced by electrolysis is supplied into the second bioreactor, while H2 from electrolysis is supplied into a tank, where it is dissolved in a third medium, a portion of which is supplied into the second bioreactor to supply H2 to the hydrogen bacteria. The main carbon source for the first process may be, for example, carbonaceous waste and by-products (e.g., biomass hydrolysates, molasses, etc.). However, other carbon sources, such as glycerol or glucose, are also possible. In this embodiment, carbon dioxide produced by yeast cells is supplied to a second bioprocess, which includes the production of biomass of cnar gas bacteria, the cnar gas bacteria growing chemosynthetically and inorganically autotrophically. Thus, in a preferred embodiment of the process of the present invention, the first type of microorganism is a microorganism of the genus Rhodosporidi, preferably Rhodosporidi torroides, and the second type of microorganism is a microorganism of the genus Hydrogenophilus, preferably Hydrogenophilus thermortheorus. Other combinations of cnar gas bacteria for the second bioprocess and yeast or bacteria for the first bioprocess are, of course, possible.

[0034] In a preferred embodiment of the process according to the present invention, the second bioprocess is carried out at a temperature between 40 and 55°C, preferably between 45 and 55°C, 50 and 55°C, or 50 and 53°C. This is particularly preferred in the case of culturing thermophilic cnar gas bacteria, such as Hydrogenophilus thermortheorus.

[0035] In a preferred embodiment of the process of the present invention, the third medium in the tank is maintained anaerobically. This may be achieved, for example, by not aerating the medium and as a result of the activity of Coenagrionis bacteria in the medium that consume all oxygen with H2 introduced into the medium.

[0036] The first, second, and third media are, in each case, aqueous media suitable for the intended purpose, for example, to support the growth of the microorganisms being cultured.

[0037] A fresh medium for the second bioprocess is preferably supplied into the tank, or may also be supplied into the second bioreactor.

[0038] In a preferred embodiment of the process of the present invention, a portion of the second medium of a second bioreactor containing a second type of microorganism is recovered from the second bioreactor, the biomass contained in the second medium is at least partially separated from the second medium, hydrolyzed, and supplied to the first bioprocess, and the second medium separated from the biomass is recycled to the second bioprocess. Alternatively, the medium separated from the biomass may be returned to the tank. In this embodiment, the biomass produced in the second bioprocess is used as an additional carbon and nitrogen source for the first bioprocess, i.e., as a substrate for growing microorganisms cultured in the first bioreactor. Furthermore, at least a portion of the biomass produced in the second bioprocess and separated from the second medium may be recovered from the entire process and used as a separate bioproduct, such as fish feed.

[0039] In a further preferred embodiment of the process of the present invention, a portion of the second medium from the second bioreactor is supplied to a tank and sprayed into the headspace of the tank containing the third medium, and a certain amount, preferably essentially the same amount as the amount of the second medium supplied to the tank, of the third medium is supplied from the tank to the second bioreactor. Since the medium in the second bioreactor and the medium in the tank are the same, a portion of the medium in the second bioreactor and the medium in the tank circulates, preferably continuously, from the second bioreactor to the tank and back to the second bioreactor. The medium is sprayed into the headspace of the tank with the help of, for example, a pump and a nozzle. The pump increases the pressure of the portion of the second medium supplied to the tank to pressure > p3, atomizing the medium in the headspace of the tank. At the same time, a certain amount, preferably the same amount, of the third medium in the tank is returned to the second bioreactor. In this embodiment, portions of the second and third media, including the bacteria contained within, circulate between the second and third bioprocesses. The fine distribution of the second medium in the headspace of the tank facilitates the transfer of H2 from the gas phase to the liquid phase. In this way, the electrolytically produced H2 gas supplied into the tank efficiently dissolves in the medium within the tank and is supplied to the second bioprocess.

[0040] The process of the present invention may be configured as a batch process, a semi-batch process, or a continuous process. Preferably, the process of the present invention is configured as a continuous process, i.e., a continuous process.

[0041] The carbon dioxide produced in the first bioprocess is recovered from the first bioreactor, for example, from the headspace of the first bioreactor, and supplied to the second bioreactor. After recovery from the first bioreactor and before introduction to the second bioreactor, the carbon dioxide is preferably separated by a suitable separation membrane, such as a hollow fiber membrane for separating N2 from CO2 (e.g., Tong Z, Sekizkardes A., "Recent Developments in High-Performance Membranes for CO2 Separation", Membranes (Basel), Vol. 11, No. 2, p. 156 (2021), doi:10.3390 / Membranes11020156; Khalilpour R., Mumford K., Zhai H., Abbas A., Stevens G., Rubin ES, "Membrane-based carbon capture from flue gas: Review"). It is concentrated by using a fluid gas (see "flue gas: a review"), Journal of Cleaner Production, Vol. 103, pp. 286-300 (2015), doi:10.1016 / j.jclepro.2014.10.050; US 10118136 B2). A suitable membrane is, for example, the hollow fiber membrane "Sepulan (registered trademark) Green" (Evonik Industrie, Essen, Germany).

[0042] The CO2 recovered from the first bioreactor may optionally be interimly stored in a storage tank, such as a gas cylinder, if, for example, the first and second bioprocesses are not proceeding in sync with respect to carbon dioxide production and consumption, and for better control of the CO2 supply to the second bioprocess.

[0043] In a preferred embodiment, oxygen produced by electrolysis is supplied directly to the second bioprocess, at least partially, to support the metabolic activity of the Kunar gas bacteria. Oxygen may be supplied to the second bioprocess continuously or intermittently. Oxygen may be stored in an intermediate storage tank, such as a gas cylinder. Oxygen may also be supplied to the second bioprocess according to physical parameters measured in the second bioprocess, such as pH, pO2, pH2, temperature, cell mass, etc. Flow rate control may be applied to control the flow rate of oxygen supplied into the second bioreactor. In this embodiment, oxygen produced by electrolysis may be supplied additionally to the first bioprocess continuously or intermittently, particularly in the case of an aerobic bioprocess. The oxygen flow rate to the first bioprocess may also be subject to control.

[0044] In a second aspect, the present invention relates to an apparatus for the biotechnological production of bioproducts by the process of the present invention, a) A first bioreactor configured to carry out a first bioprocess, the first bioprocess comprising culturing a first type of microorganism in a first medium at a first pressure p1, the first bioprocess resulting in the production of bioproducts and carbon dioxide CO2 in the first bioreactor, b) A second bioreactor configured to carry out a second bioprocess, the second bioprocess comprising culturing a second type of microorganism in a second medium at a second pressure p2, the second bioprocess being a process that produces biomass and consumes molecular oxygen O2, molecular hydrogen H2, and at least a portion of the CO2 produced in the bioprocess of step a), c) A tank configured to house the headspace and a third medium at a third pressure p3, d) An electrolysis apparatus for electrolyzing water into O2 and H2, The device includes, and further, - For the transfer of CO2 produced in the first bioreactor to the second bioreactor, a first fluid connection part is provided to fluidly connect the first outlet of the first bioreactor to the first inlet of the second bioreactor, - For the transfer of biomass produced in the second bioreactor to the first bioreactor, a second fluid connection part is provided to fluidly connect the first outlet of the second bioreactor to the first inlet of the first bioreactor, - For the transfer of the second medium to the tank, a third fluid connection part is provided to fluidly connect the second outlet of the second bioreactor to the first inlet of the tank, - For the transfer of the third medium to the second bioreactor, a fourth fluid connection is provided, which fluidly connects the first outlet of the tank to the second inlet of the second bioreactor. - A fifth fluid connection part that fluidly connects the first outlet of the electrolysis device to the third inlet of the second bioreactor for the transfer of oxygen O2 to the second bioreactor, - A sixth fluid connection part that fluidly connects the first outlet of the electrolysis device to the third inlet of the first bioreactor for the transfer of oxygen O2 to the first bioreactor, - For the transfer of hydrogen H2 to the tank, a seventh fluid connection is provided to fluidize the second outlet of the electrolysis apparatus to the second inlet of the tank, Regarding devices including...

[0045] The apparatus of the present invention is configured to carry out the process of the present invention and includes a first bioreactor, a second bioreactor, and a tank which may be configured as a bioreactor. These bioreactors and tank are fluidically connected to one another to carry out the process of the present invention. The apparatus thus includes fluid connections, such as pipes or tubes or similar, that fluidly interconnect these bioreactors and tank to enable the transfer of CO2 produced in the first bioreactor to the second bioreactor, the transfer of biomass from the second bioreactor to the first bioreactor, the supply of oxygen generated by an electrolysis device to the second bioreactor and / or the first bioreactor, the supply of hydrogen to the tank, and the circulation of a medium between the second bioreactor and the tank.

[0046] In a preferred embodiment of the apparatus of the present invention, the second fluid connection includes, between the first outlet of the second bioreactor and the first inlet of the first bioreactor, a separation device for separating a second type of microorganism from the second medium, and a hydrolysis device for hydrolyzing the second type of microorganism separated from the second medium by the separation device. The separation device may be a centrifuge or hollow fiber membrane module operating in batch or continuous mode. The medium separated from the second type of microorganism may be returned to the second bioreactor and supplied into the tank or recovered from the process.

[0047] In a further preferred embodiment of the apparatus of the present invention, a third fluid connection portion, which fluidly connects the second outlet of the second bioreactor to the first inlet of the tank, extends into the upper part of the tank, including the headspace, and the extended portion includes a nozzle for atomizing the second medium supplied into the tank.

[0048] More preferably, the first fluid connection, which fluidly connects the first outlet of the first bioreactor to the first inlet of the second bioreactor, includes a CO2 concentrator, preferably a membrane-based CO2 concentrator, for concentrating the CO2 produced in the first bioreactor and recovered from the first bioreactor before it is transferred to the second bioreactor. Even more preferably, the first fluid connection includes a CO2 storage tank, such as a gas cylinder, for storing the CO2. Thus, the CO2 may be interimly stored and supplied to the second bioprocess when needed. This is advantageous because it allows for the decoupling of CO2 production in the first bioprocess from CO2 consumption in the second bioprocess, for example, in the event of process failure or to compensate for unequal CO2 production and consumption rates.

[0049] A sixth fluid connection, which fluid-connects the first outlet of the electrolysis apparatus to the third inlet of the first bioreactor for the transfer of oxygen O2 to the first bioreactor, shares a portion of the fifth fluid connection. In a preferred embodiment of the apparatus of the present invention, the fifth fluid connection, which fluid-connects the first outlet of the electrolysis apparatus to the third inlet of the second bioreactor, extends into the first bioreactor and fluid-connects the first outlet of the electrolysis apparatus to the third inlet of the first bioreactor. Preferably, a two-way valve is inserted in the fifth fluid connection so that the oxygen flow can be divided and directed to both the first and second bioreactors simultaneously, or intermittently directed to either the first or second bioreactor.

[0050] The tank may be configured as a bioreactor so that the apparatus includes first, second, and third bioreactors.

[0051] In the following, the present invention will be described in more detail with reference to the accompanying drawings, which are merely examples. [Brief explanation of the drawing]

[0052] [Figure 1]A schematic diagram showing an embodiment of the apparatus according to the present invention. Gas flow is represented by a dashed line, and liquid flow by a solid line. [Figure 2] A schematic diagram showing the gas flow and liquid flow in an embodiment of the apparatus according to the present invention shown in Figure 2. The gas flow is represented by a dashed line, and the liquid flow by a solid line. [Modes for carrying out the invention]

[0053] Figure 1 shows a simplified schematic diagram of apparatus 1 of the present invention. The apparatus includes three compartments A, B, and C. Compartment A includes a first bioreactor 2, compartment B includes a second bioreactor 3, and compartment C includes a tank 4 which may be configured as a third bioreactor. The first bioreactor 2 and the second bioreactor 3 are equipped with a first agitator 21 and a second agitator 31, respectively, for agitating the first medium 23 in the first bioreactor 2 or the second medium 33 in the second bioreactor 3. A first pressure p1 is dominant in the first bioreactor 2, a second pressure p2 is dominant in the second bioreactor 3, and a third pressure p3 is dominant in the tank 4.

[0054] In compartment A, which includes a first bioreactor 2, a first bioprocess is carried out, in which a desired biochemical substance is produced by a first type of microorganism cultured in the first bioreactor 2, and the production of carbon dioxide (CO2) is involved. An example of such a first bioprocess is aerated yeast fermentation by Rhodosporidia tolloides for the production of variable lipids. Here, for example, carbon-containing waste materials and by-products (biomass hydrolysates, molasses, etc.) are used as the main carbon source. In addition to lipids, R. tolloides is used as feed in aquaculture (shrimp farms, salmon farms, e.g., Blomqvist, J., Pickova, J., Tilami, SK et al., "Oleaginous yeast as a component in fish feed," Scientific Reports, Vol. 8, p. 15945 (2018), doi:10.1038 / s41598-018-34232-x) or as a coloring agent in the food industry (e.g., Igreja, WS, Maia, FdA, Lopes, AS, Christe, RC, "Biotechnological Production of Carotenoids Using Low Substrates is Affected by Culture Parameters" - Review Article). "Cost-Substrates Is Influenced by Cultivation Parameters: A Review," International Journal of Molecular Sciences, Vol. 22, p. 8819 (2021), see doi:10.3390 / ijms22168819, which describes the production of carotenoid-containing biomass that can be used.The lipid profile can be modified by genetic engineering of R. toruloides (Wen Z., Zhang S., Odoh CK, Jin M., Zhao ZK, "Rhodosporidium toruloides—A potential red yeast chassis for lipids and beyond," FEMS Yeast Research, vol. 20, no. 5, p. foaa038 (2020), doi:10.1093 / femsyr / foaa038). The produced lipid profiles can be modified and adapted to classify their lipids for the cosmetics / food industry, for example, from palm oil-like to cocoa butter-like.

[0055] Air 100 is passed through a first bioreactor 2 containing a first medium 23 and cells of a first type of microorganism, in this case for example, the oily yeast Rhodosporidia tolloides. Alternatively, oxygen generated in an electrolysis unit 5 that electrolyzes water may be supplied to the first bioreactor 2. The exhaust gas containing CO2 produced in the first bioreactor 2 is recovered from the headspace 32 of the first bioreactor 2 using a compressor 9 and supplied as a CO2 source to a bioprocess carried out in the second bioreactor 3 in compartment B via a first fluid connection 61. The first fluid connection 61, which may be a pipe or tube, fluidly connects the first outlet 25 of the first bioreactor 2 to the first inlet 34 of the second bioreactor 3. On its way to the second bioreactor 3, the CO2 is concentrated using a CO2 concentrator 8, for example, a membrane-based CO2 concentrator 8, so that CO2-deficient air 102, mainly containing N2, is removed from the exhaust flow. Therefore, the air supply rate vg2 (see Figure 2) at which CO2, or more precisely, CO2-enriched air, is supplied to the second bioreactor 3 is set by the airflow rate, i.e., the rate at which air 100 is supplied to the first bioreactor 2 and compressor 9. In the illustrated embodiment, the first fluid connection 61 further includes a storage tank 13 for intermediate storage of CO2 after the CO2 concentrator 8 in the direction of the CO2 flow. Bioproducts 101, for example, biochemicals produced in compartment A, such as chemicals discharged into the first medium 23 in the first bioprocess or accumulated in the proliferated cells, can be recovered from the bioprocess in compartment A.

[0056] In compartment B, a second type of microorganism, namely chemosynthetic inorganic nutrient-rich so-called "Knurl gas bacteria," grows in the second bioreactor 3, dependent on CO2, H2, and O2. The second type of microorganism cultured in compartment B may be any microorganism having the specified gas requirements. A suitable example of a microorganism is Hydrogenophilus thermoluteolus, which has a particularly high growth rate (Arai H, Shomura Y, Higuchi Y, Ishii M, "Complete Genome Sequence of a Moderately Thermophilic Facultative Chemolithoautotrophic Hydrogen-Oxidizing Bacterium, Hydrogenophilus thermoluteolus TH-1", Microbiology Resource Announcements, Vol. 7, No. 6, pp. e00857-18 (2018), doi:10.1128 / MRA.00857-18). The bioprocess in section B receives CO2 and residual O2 from the related bioprocess in section A, as well as pure H2 and O2 from the electrolysis unit 5, which is described in more detail below.

[0057] O2 and H2 used by hydrogen bacteria growing in the second bioreactor 3 are generated by the electrolysis device 5. O2 is supplied directly to the second bioreactor via a fifth fluid connection 65 that fluidly connects the first outlet 55 of the electrolysis device 5 to the third inlet 38 of the second bioreactor 3. In an embodiment of the apparatus 1 of the present invention shown in Figure 1, an extended portion 651 of the fluid connection 65 fluidly connects the third inlet 28 of the first bioreactor 2 to the fluid connection 65, and therefore to the first outlet 55 of the electrolysis device 5, so that O2 generated by the electrolysis device 5 can also be supplied to the first bioreactor 2 via a sixth fluid connection 66, the sixth fluid connection 66 sharing a portion of the fifth fluid connection 65. A two-way valve 86 may be used to divide the O2 gas flow into a first portion vg4 (see Figure 2) supplied to the second bioreactor 3 and a second portion vg5 supplied to the first bioreactor 2 via an extended portion 651. Alternatively, the valve 86 may be used to direct the O2 flow only to the first bioreactor 2, or to intermittently direct the O2 flow to either the second bioreactor or the first bioreactor 2. The O2 gas may be interimly stored in a separate storage tank (not shown).

[0058] Due to the significantly lower solubility of H2 in the aqueous medium used, and to avoid a flammable space and thus create explosion protection by separating the gas phase containing O2 or H2, H2 is not supplied directly to the second bioreactor 3. Instead, the H2 inlet is relocated to compartment C, which includes a tank 4 containing a third medium 43, where the third medium 43 corresponds to the second medium 33 in the second bioreactor 3. The H2 gas is supplied to tank 4 via a seventh fluid connection 67 that fluidly connects the second outlet 57 of the electrolysis unit 5 to the second inlet 46 of tank 4. The H2 gas may also be intermediately stored in a separate storage tank (not shown). The electrolysis unit 5 accumulates excess pressure in tank 4. In this embodiment, the dominant pressure p3 in tank 4 is the highest pressure compared to the pressures p1 and p2 in the other compartments, namely compartments A and B. The increase in pressure provides a higher solubility coefficient of H2. The pressure is controlled by the current I (proportional to the H2 gas supply rate vg3, see Figure 2) flowing through the electrolysis apparatus 5 and the position (closed) of the second valve 82. The medium circulating between compartment B and compartment C has an additional favorable effect on the introduction of H2 gas. For this medium cycle operation, third and fourth fluid connections 63 and 64 are located between the second bioreactor 3 and the tank 4. The third fluid connection 63 fluidly connects the second outlet 37 of the second bioreactor 3 to the first inlet 44 of the tank, which is located at the top 41 of the tank 4 at the height of the tank's headspace 42. A first pump 10 is located in the third fluid connection 63 to pump the second medium 33 from the second bioreactor 3 to the tank 4. The fourth fluid connection section 64 fluidly connects the first outlet 45 of the tank 4 to the second inlet 36 of the second bioreactor 3, allowing the return flow of the third medium 43 from the tank 4 to the second bioreactor 3. The flow of the third medium 43 to the second bioreactor 3 can be controlled, for example, by the second valve 82. A portion of the second medium 33 is recovered from compartment B and pressurized >p3 using the first pump 10 to finely spray it through the nozzle 12 into the headspace 42 of the tank 4 in compartment C. This fine distribution into the headspace 42 of the tank 4, which has an H2 atmosphere, ensures further concentration of H2 in the liquid phase.The inflow of H2-saturated medium into the second bioprocess in section B can be regulated by a circuit consisting of the first pump 10 and the second valve 82 using vl1 and vl2 (see Figure 2). During this cycle, there may also be a second type of microorganism that can benefit from various dissolved gas concentrations, namely, Kunar gas bacteria.

[0059] Compartment B preferably has a pressure p2 lower than pressure p3, where p2 allows dissolved H2 to be released more effectively into the surrounding medium. Here, the inflow of CO2 and O2 is increased by stirring by a second agitator 31 and by an optional gas cycle between compartment A and compartment B via a valve 81 located in a ninth fluid connection 69 that fluidly connects the third outlet 39 of the second bioreactor 3, located at the height of the headspace 32 of the second bioreactor 3, and the second inlet 26 of the first bioreactor 2, located at the height of the headspace 22 of the first bioreactor 2.

[0060] The pressure p1 in compartment A is preferably lower than in compartments B and C, but preferably slightly above atmospheric pressure. In compartments B and C, if necessary, excess pressure can be reduced or controlled using valves 83 and 84.

[0061] The biomass generated in section B is preferably continuously withdrawn via a second pump 11 and a separation device 6 once it reaches a sufficient cell density of the second type of microorganism. In this process, the second medium 33 containing the second type of microorganism is recovered from the second bioreactor 3, and the cells are separated from the second medium 33 using a separation device 6, such as a continuous centrifuge, such as a disc stack centrifuge, or a hollow fiber membrane module, hydrolyzed in a hydrolysis device 7, and supplied to the first bioreactor 2. For this purpose, a second fluid connection 62, such as a pipe or tube, fluidly connects the first outlet 35 of the second bioreactor 3 to the first inlet 24 of the first bioreactor 2. The separation device 6 is positioned after the pump 11 with respect to the direction of flow of the second medium 33, and the hydrolysis device 7 is positioned after the separation device 6. In this embodiment, the second medium 33 separated from the cells is returned to the bioprocess in compartment B via an eighth fluid connection 68 that fluidly connects the portion of the separation device 6 containing the separated second medium 33 to the fourth inlet 30 of the second bioreactor 3. Optionally, the second medium may be removed from the circuit via a fifth valve 85 for reprocessing, for example, and reintroduced into compartment C. The extracted biomass is hydrolyzed using a hydrolyzer 7 and supplied to the bioprocess in compartment A as an additional C and N source. At least a portion of the biomass may also be recovered and used as a separate bioproduct, such as fish feed. Depending on the operating mode of the bioprocess in compartment A (e.g., batch or continuous), the biomass post-processing may be performed continuously or at intervals. In this way, an estimated 20-60% of the expensive glucose in the lipid fermentation step can be saved, and the C balance can be significantly shifted toward higher lipid yields.

[0062] In section C, for example, fresh medium 103 may be introduced into the process at the third inlet 48 of tank 4. For example, a first type of microorganism and / or the first medium 23 may be recovered from section A via the third outlet 29 of the first bioreactor 2.

[0063] Figure 2 shows a simplified configuration of the apparatus shown in Figure 1, illustrating the gas flow (dashed lines) and liquid flow (solid lines) to and between compartments A, B, or C (see description in Figure 1). [Explanation of Symbols]

[0064] 1. Apparatus of the present invention 2. First Bioreactor 21 First stirring device 22 headspace 23 First medium 24 First entrance 25 First Exit 26. The second entrance 28 The Third Entrance 29 Third Exit 3. Second Bioreactor 30 The fourth entrance 31. Second stirring device 32 headspace 33 Second medium 34. The first entrance 35 First Exit 36. The second entrance 37. Second Exit 38 The Third Entrance 39 Third Exit 4 tanks 41 Top of tank 42 headspace 43 The third medium 44 First entrance 45 First Exit 46. ​​The second entrance 48 The Third Entrance 5. Electrolysis apparatus 55 (First) Exit 57 (Second) Exit 6 Separation device 61 First fluid connection 62 Second fluid connection 63 Third fluid connection 64 Fourth fluid connection 65 Fifth fluid connection 651 Extended part 66 Sixth fluid connection 67 Seventh fluid connection 68. Eighth fluid connection 69. Ninth fluid connection 7 Hydrolysis equipment 8 CO2 concentrator 81 valves 82 Second valve 83 valves 84 valves 85 The fifth defense 86 Two-way valve 9 Compressor 10. First pump 11. Second pump 12 nozzles 13 CO2 storage tanks 100 air 101 Bioproducts 102 CO2-deficient air 103 Fresh medium p1 First pressure p2 Second pressure p3 Third pressure

Claims

1. A process for the biotechnological production of bioproducts, a) comprising culturing a first type of microorganism in a first medium at a first pressure p1 in a first bioreactor, wherein the bioproduct and carbon dioxide CO ２ A first bioprocess that results in the generation of, wherein the carbon dioxide produced in the first bioprocess is supplied to the second bioprocess. b) A second bioprocess comprising culturing a second type of microorganism in a second medium at a second pressure p2 in a second bioreactor, wherein biomass is produced and molecular oxygen O ２ and molecular hydrogen H ２ and the CO produced in the first bioprocess of step a) ２ A second bioprocess which is a process that consumes at least a portion of and c) Water O ２ and H ２ The O2O2 is produced by electrolysis. ２ At least a portion of is supplied to the second bioprocess and / or the first bioprocess, and H produced by electrolysis ２ The steps include supplying at least a portion of the headspace and a tank containing a third medium at a third pressure p3, wherein the third medium is identical to the second medium in the second bioreactor, d) H ２ supplying at least a portion of said third medium containing from said tank to said second bioprocess; This includes, and at least a portion of the biomass produced in the second bioprocess of step b) is used as a C source for the first bioprocess of step a), The first type of microorganism is a yeast cell, the second type of microorganism is a conditionally chemosynthetic inorganic autotrophic hydrogen bacterium, and the second bioprocess is O ２ , H ２ and CO ２ Includes chemically synthesized inorganic autotrophic consumption, The first medium, the second medium, and the third medium are, in each case, aqueous media suitable for supporting the growth of microbial cultures in the process.

2. The process according to claim 1, wherein p3 ≥ p2 ≥ p1.

3. The process according to claim 1 or 2, wherein the first bioprocess is an aerobic bioprocess.

4. The process according to any one of claims 1 to 3, wherein the second bioprocess is carried out at a temperature between 40 and 55°C.

5. The process according to any one of claims 1 to 4, wherein a portion of the second medium of the second bioreactor containing the second type of microorganisms is recovered from the second bioreactor, the biomass contained in the second medium is at least partially separated from the second medium, hydrolyzed and supplied to the first bioprocess, and the second medium separated from the biomass is recycled to the second bioprocess.

6. The process according to any one of claims 1 to 5, wherein a portion of the second medium from the second bioreactor is supplied into the tank and sprayed into the headspace of the tank, and the same amount of the third medium is supplied from the tank to the second bioreactor.

7. O produced by electrolysis ２ A portion of the process according to any one of claims 1 to 6 is supplied to the first bioprocess.

8. The process according to any one of claims 1 to 7, wherein the first type of microorganism is a microorganism of the genus Rhodosporidium, and the second type of microorganism is a microorganism of the genus Hydrogenophilus.

9. The process according to any one of claims 1 to 8, wherein p1 is greater than atmospheric pressure.

10. The process according to any one of claims 1 to 9, wherein the process is a continuous process.

11. Apparatus (1) for the biotechnological production of a bioproduct by a process according to any one of claims 1 to 10, a) A first bioreactor (2) configured to carry out a first bioprocess, the first bioprocess comprising culturing a first type of microorganism in a first medium (23) at a first pressure p1, the first bioprocess comprising the bioproduct and carbon dioxide CO ２ The first bioreactor (2) is the result of the generation of the following b) A second bioreactor (3) configured to carry out a second bioprocess, the second bioprocess comprising culturing a second type of microorganism in a second medium (33) at a second pressure p2, the second bioprocess producing biomass and molecular oxygen O ２ and molecular hydrogen H ２ And the CO produced in the bioprocess of step a) ２ A second bioreactor (3) is a process that consumes at least a portion of the and c) A tank (4) configured to house a headspace (42) and a third medium (43) at a third pressure p3, d) Water O ２ and H ２ An electrolysis apparatus (5) for electrolysis, The apparatus (1) further includes, - CO produced in the first bioreactor (2) ２ For the transfer of to the second bioreactor (3), a first fluid connection part (61) is provided which fluidly connects the first outlet (25) of the first bioreactor (2) to the first inlet (34) of the second bioreactor (3), - For the transfer of biomass produced in the second bioreactor (3) to the first bioreactor (2), a second fluid connection part (62) is provided, which fluidly connects the first outlet (35) of the second bioreactor (3) to the first inlet (24) of the first bioreactor (2), - For the transfer of the second medium (33) to the tank (4), a third fluid connection part (63) is provided, which fluidly connects the second outlet (37) of the second bioreactor (3) to the first inlet (44) of the tank (4), - For the transfer of the third medium (43) to the second bioreactor (3), a fourth fluid connection part (64) is provided, which fluidly connects the first outlet (45) of the tank (4) to the second inlet (36) of the second bioreactor (3), - Oxygen O ２ For the transfer of the fluid to the second bioreactor (3), a fifth fluid connection part (65) is provided, which fluidly connects the first outlet (55) of the electrolysis apparatus (5) to the third inlet (38) of the second bioreactor (3), - Oxygen O ２ For the transfer of the first bioreactor (2), a sixth fluid connection part (66) is provided, which fluidly connects the first outlet (55) of the electrolysis apparatus (5) to the third inlet (28) of the first bioreactor (2), - Hydrogen H ２ For the transfer of to the tank (4), a seventh fluid connection part (67) is provided, which fluidly connects the second outlet (57) of the electrolysis apparatus (5) to the second inlet (46) of the tank (4), Includes, Apparatus (1), wherein the first medium, the second medium, and the third medium are, in each case, aqueous media suitable for supporting the growth of microbial cultures.

12. The apparatus (1) according to claim 11, wherein the second fluid connection (62) includes, between the first outlet (35) of the second bioreactor (3) and the first inlet (24) of the first bioreactor (2), a separation device (6) for separating the second type of microorganism from the second medium (33) and a hydrolysis device (7) for hydrolyzing the second type of microorganism separated from the second medium (33) by the separation device (6).

13. The apparatus (1) according to claim 11 or 12, wherein the third fluid connection portion (63) fluidly connects the second outlet (37) of the second bioreactor (3) to the first inlet (44) of the tank (4), extends into the upper part (41) of the tank (4), including the headspace (42), and the extended portion includes a nozzle (12) for atomizing the second medium (33) supplied to the tank (4).

14. The first fluid connection part (61) that fluidly connects the first outlet (25) of the first bioreactor (2) to the first inlet (34) of the second bioreactor (3) is CO ２ CO to be concentrated before its transfer to the second bioreactor (3) ２ Apparatus (1) according to any one of claims 11 to 13, comprising a concentration device (8).

15. The first fluid connection part (61) that fluidly connects the first outlet (25) of the first bioreactor (2) to the first inlet (34) of the second bioreactor (3) is CO ２ CO2 for intermediate storage ２ Apparatus (1) according to any one of claims 11 to 14, including a storage tank (13).

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