GB2611124A

GB2611124A - Method

Info

Publication number: GB2611124A
Application number: GB2201454.2A
Authority: GB
Inventors: Patrick Kincaid Kevin; Luis Busi Da Silva Marcio; Karimi Mojtaba; Amanda Goncalves Renata; Allen Harris Roger; Christopher Trevino Aaron; Richard Broussard Zachary; Karimi Tahereh; Lesse Andrade Lahme Luiza; De Freitas Magalhaes Barbara; Walter Rimbau Christian
Original assignee: Cemvita Factory Inc
Current assignee: Cemvita Factory Inc
Priority date: 2021-09-24
Filing date: 2022-02-04
Publication date: 2023-03-29
Also published as: GB2611370A; GB2611123A; GB2613039A

Abstract

The present invention provides a method of increasing production of a hydrogen containing liquid, such as methanol, ammonia, ammonium or C2-C120 hydrocarbons from an enclosed bioreactor, wherein the bioreactor is a subterranean formation or landfill enclosure, comprising: providing a baseline reaction mixture in the enclosed bioreactor, wherein the mixture includes a substrate, water, and at least one microorganism; producing baseline microorganism data on an identity and percentage of the at least one microorganism, relative to a total percentage of microorganisms in the baseline reaction mixture; measuring an amount of hydrogen-containing liquid in a sample collected from the enclosed bioreactor; increasing production of the hydrogen containing liquid from the enclosed bioreactor by forming a synthetic reaction mixture, comprising additional microorganisms or a palladium catalyst, and harvesting the hydrogen-containing liquid from the enclosed bioreactor by separating the hydrogen-containing liquid from solids and other liquids by transferring the hydrogen-containing liquid into a storage container.

Description

METHOD

TECHNICAL FIELD

The present invention concerns a method of, and system for, increasing production of a hydrogen-containing liquid from an enclosed bioreactor

BACKGROUND

Hydrogen is an important fuel and chemical process substrate. It is known in the art to use microbes to produce hydrogen from hydrocarbon substrates.

US20070298479 describes a process for stimulating microbial hydrogen production in a petroleum-bearing subterranean formation, comprising: (a) analyzing one or more components of the formation to determine characteristics of the formation environment; (b) detecting the presence of a microbial consortium, comprising at least one fermentative syntrophic microorganism, within the formation; (c) assessing whether the formation microorganisms are currently active; (d) determining whether the microbial consortium comprises one or more fermentative syntrophic microorganisms; (e) characterization of one or more fermentative syntrophic microorganism of the consortium, and comparing the one or more characterized organisms with at least one known characterized known microorganism having one or more known physiological and ecological characteristics; (f) characterizations of one or more hydrogen consuming microorganism of the consortium, and comparing the one or more characterized microorganisms with at least one known characterized, known microorganism having one or more known physiological and ecological characteristics; (g) using information obtained from steps (a) through (e) for determining an ecological environment that promotes in situ microbial degradation of petroleum and promotes microbial generation of hydrogen by at least one fermentative syntrophic microorganism of the consortium; (h) using information obtained from steps (a) and (f), if methanogenic, sulphate reducing, or other hydrogen consuming microorganisms are present, for determining an ecological environment that inhibits in situ microbial degradation of hydrogen by at least one hydrogen-oxidizing microorganism of the consortium; (i) modifying the formation environment based on the determinations of step (g) and (h), if methanogenic sulphate reducing or other hydrogen-oxidizing microorganisms are present, to stimulate microbial conversion of petroleum to hydrogen; and (j) removal of the hydrogen generated from the sites of generation.

However, there is a need in the art for improved methods of increasing the production of clean hydrogen.

SUMMARY

According to a first aspect of the present invention, there is provided a method of increasing production of a hydrogen-containing liquid from an enclosed bioreactor comprising: providing a baseline reaction mixture in the enclosed bioreactor, wherein the baseline reaction mixture includes a substrate, water, and a baseline amount of at least one microorganism, wherein the substrate includes a nitrogen source, an unsaturated hydrocarbon having from 2 to 120 carbon atoms, methane, hydrogen, or a combination thereof, wherein the hydrogen-containing liquid includes ammonia, ammonium, methanol, a saturated hydrocarbon having from 2 to 120 carbon atoms, or a combination thereof; producing baseline microorganism data on an identity and a baseline percentage of the at least one microorganism, relative to a baseline total percentage of microorganisms in the baseline reaction mixture, by performing DNA and/or RNA sequencing of a baseline microorganism sample from the baseline reaction mixture; measuring a baseline amount of hydrogen-containing liquid in a baseline sample collected from the enclosed bioreactor; increasing production of the hydrogen-containing liquid from the enclosed bioreactor by forming a synthetic reaction mixture, and harvesting the hydrogen-containing liquid from the enclosed bioreactor at a hydrogen-containing liquid harvesting rate by separating the hydrogen-containing liquid from solids and other liquids by transferring the hydrogen-containing liquid into a hydrogen-containing liquid storage container. The synthetic reaction mixture is formed by: adding at least one non-native hydrogen-containing liquid producing microorganism until a percentage of the non-native hydrogen-containing liquid producing microorganism in the synthetic reaction mixture is at least 20% of a total amount of microorganisms in the synthetic reaction mixture; or adding at least one hydrogen-containing liquid production enhancer or a palladium-containing catalyst to the baseline reaction mixture until a post-baseline amount of hydrogen-containing liquid in a post-baseline sample collected from the enclosed bioreactor is at least 10% higher than the baseline amount of hydrogen-containing liquid; or adding at least one recombinant microorganism to the baseline reaction mixture until a percentage of the at least one recombinant microorganism in the synthetic reaction mixture is at least 20% of a total amount of microorganisms in the reaction mixture, or a combination thereof. The enclosed bioreactor is a bioreactor subterranean formation, a bioreactor landfill enclosure, or a combination thereof.

The nitrogen source may include nitrogen gas, agriculture waste, soy protein isolate, blood meal, feather meal, dried fish, yeast extract, nitrates, nitrites, urea, soy flour, peanut cake, peptone, beef extract, or a combination thereof.

The method may further comprise after providing the baseline reaction mixture, but before forming the synthetic reaction mixture, producing baseline environmental data from the baseline reaction mixture. The baseline environmental data may include one or more of the following measurements of a baseline environmental sample from the baseline reaction mixture: pH; temperature; water analysis; oxidation-reduction potential; pressure; dissolved oxygen; hydrocarbon concentrations; volatile fatty acids concentrations; cation concentration; anion concentration; concentration of gases (such as one or more of NH3, CO2, CO, H2, H2S and CH4; salt concentration; and metal concentration..

The baseline microorganism sample and the baseline environmental sample may be the same or different.

In some embodiments, the substrate includes methane and the hydrogen-containing liquid may include methanol. In some embodiments, the at least one non-native hydrogen-containing liquid producing microorganism and/or the at least one recombinant microorganism has a genus of Methanothermobacter, Methanosaeta, Methanococcales, Candidatus, Desulfobulbus, Desulfocapsa, Desulfofustis, Desulfopila, Desulforhophalus, Desulfotalea, Desulfurivibrio, Methylomirabilis, uncultured anaerobic Methanotrophic archaea (ANME-1), uncultured ANME-2, uncultured ANME-3, or combination or mixture thereof.

In some embodiments, the substrate includes a nitrogen source and the hydrogen-containing liquid includes ammonia or ammonium. In some embodiments, the at least one non-native hydrogen-containing liquid microorganism and/or the at least one recombinant microorganism has a genus of Methanothermobacter, Methanosaeta, Methanococcales, Candidatus, Desulfobulbus, Desulfocapsa, Desulfofustis, Desulfopila, Desulforhophalus, Desulfotalea, Desulfurivibrio, Methylomirabilis, uncultured anaerobic Methanotrophic archaea (ANME-1), uncultured ANME-2, uncultured ANME-3 or a family of Methanoperedenaceae, Methanobacteriaceae, Methanocaldococcaceae, Desuffobufbaceae, Candidatus Methanoperedenaceae.

The at least one non-native hydrogen-containing liquid producing microorganism or the at least one recombinant microorganism may be the same or different.

The recombinant microorganism may express at least one Coenzyme M reductase and or dehydrogenase protein having a gene sequences at least 95% identical to SEQ ID NO.

[mmg:MTBMA_c15480], [mth:MTH_1015], [mmg:MTBMA_c15520], [mmg:MTBMA_c15490], [mth:MTH_1166], [mth:MTH_1167], [eco:b4346], [eco:b4345], [ag:AAA22593], [mea:Mex_1p4538, [mea:Mex_1p4535], [ag:ACS29499], [ag:CAH55641], [mrd:Mrad2831_0508], by expressing a non-native Coenzyme M reductase and or dehydrogenase expressing nucleotide sequence.

Preferably, an amount of hydrogen-containing liquid produced or protein produced by the recombinant microorganism is greater than that produced relative to a control microorganism lacking the non-native Coenzyme M reductase and or dehydrogenase expressing nucleotide sequences.

The substrate may include the saturated hydrocarbon having from 2-70 carbon atoms, or 2 to 40 carbon atoms.

The hydrogen-containing liquid may include the saturated hydrocarbon having from 2-70 carbon atoms, or 2 to 40 carbon atoms.

The palladium-containing catalyst may include palladium on carbon, palladium on barium carbonate, palladium on barium sulfate, palladium on calcium carbonate, palladium on silica gel, Pd(OH)2, or a combination or a mixture thereof.

The hydrogen-containing liquid harvesting rate may be at least about 0.1 Uhr, or at least about 1 L/hr, or at least about 10 Uhr, or at least about 100 Uhr. The hydrogen harvesting rate may be up to about 106 Uhr, or up to about 105 Uhr, or up to about 104 L/hr, or up to about 103 L/hr. The hydrogen harvesting rate may be from about 0.1 Uhr to about 106 Uhr, or from about 0.1 L/hr to about 103 Uhr, or from about 103 L/hr to about 106 L/hr.

The unsaturated hydrocarbon having from 2 to about 120 carbon atoms may include an alkene, an alkyne, an aromatic hydrocarbon, or a polyaromatic hydrocarbon.

The bioreactor subterranean formation may include a natural formation, non-natural formation, a hydrocarbon-bearing formation, a natural gas-bearing formation, a methane-bearing formation, a depleted hydrocarbon formation, a depleted natural gas-bearing formation, a wellbore, or a combination thereof.

The bioreactor landfill enclosure may include a landfill that is enclosed by a building material. The building material may include at least one of a brick, a cement, a plastic, a non-natural rubber, a geomembrane of any kind, concrete, steel, a glass, or a combination thereof.

The hydrogen-containing liquid production enhancer may be a biocidal inhibitor, a methanogenesis inhibitor, a sulfate reduction inhibitor, a nitrate reduction inhibitor, an iron reduction inhibitor, or a combination thereof.

The biocidal inhibitor may be glutaraldehyde, a quaternary ammonium compound, formaldehyde, a formaldehyde releaser such as 3,3'-methylenebis[5-methyloxazolidine], dibromonitrilopropionamide, tetrakis hydroxymethyl phosphonium sulfate, chlorine dioxide, peracetic acid, tributyl tetradecyl phosphonium chloride, methylisothiazolinone, chloromethylisothiazolinone, sodium hypochlorite, dazomet, dimethyloxazolidine, trimethyloxazolidine, N-bromosuccinimide, bronopol, or 2-propenal, or a mixture thereof.

The methanogenesis inhibitor may be bromethane sulfonic acid, an aminobenzoic acid, 2-bromoethanesulfonate, 2-chloroethanesulfonate, 2-mercaptoethanesulfonate, lumazine, a fluoroacetate, nitroethane, or 2-nitropropanol, or a mixture thereof.

The sulfate reduction inhibitor may be a molybdate salt, a nitrate salt, a nitrite salt, a chlorate salt, or a perchlorate salt or a mixture thereof.

The nitrate reduction inhibitor may be sodium chlorate, a chlorate salt, or a perchlorate salt, or a mixture thereof.

Forming the synthetic reaction mixture may include one or more of: a) adding the at least one non-native hydrogen-containing liquid producing microorganism until a percentage of the non-native hydrogen-containing liquid producing microorganism in the synthetic reaction mixture is at least 51% of a total amount of microorganisms in the synthetic reaction mixture; b) adding the at least one hydrogen-containing liquid production enhancer or the palladium-containing catalyst to the baseline reaction mixture until a post-baseline amount of hydrogen-containing liquid in a post-baseline gas sample of liquids collected from the enclosed bioreactor is at least 51% higher than the baseline amount of hydrogen-containing liquid; and/or c) adding the at least one recombinant microorganism to the baseline reaction mixture until a percentage of the at least one recombinant microorganism in the synthetic reaction mixture is at least 51% of a total amount of microorganisms in the reaction mixture.

The method may further comprise harvesting hydrogen from the enclosed bioreactor at a hydrogen harvesting rate, and separating the hydrogen from other gasses by filtering the hydrogen through a hydrogen-selective membrane filter and transferring the hydrogen into a hydrogen storage container.

According to a second aspect of the present invention, there is provided a system for increasing production of a hydrogen-containing liquid from an enclosed bioreactor comprising the enclosed bioreactor, a hydrogen-containing liquid storage container, and a hydrogen-containing liquid separator. The reaction mixture includes a substrate, water, and a baseline amount of at least one microorganism. The hydrogen-containing liquid separator includes at least one hydrogen-containing liquid membrane filter, at least one solids filter, or at least one distillation apparatus. The enclosed bioreactor is connected to the hydrogen-containing liquid separator by a hydrogen-containing liquid path. The hydrogen-containing liquid separator is connected to the hydrogen-containing liquid container by a purified hydrogen-containing liquid path.

The enclosed bioreactor may have a volume of at least about 100 rn3, or at least about 103 m3, or at least about 104 m3, or at least about 105 m3. The enclosed bioreactor may have a volume of up to about 4 x 109 m3, or up to about 4 x 108 m3, or up to about 4 x 10 rn3, or up to about 4 x 106 m3. The enclosed bioreactor may have a volume of from about 100 m3 to about 4 x 109 m3, or from about 100 rri3 to about 4 x 106 m3, or from about 4 x 106 m3 to about 4 x 10° m3.

The hydrogen-containing liquid storage container may include a liquid tank or a hydrogen-containing liquid surface formation.

The system may further comprise a genetic material testing facility, preferably within about 1000 meters of a resealable opening of the enclosed bioreactor and connected to the resealable opening by a genetic material testing liquid pathway. The genetic material testing facility may contain at least one DNA and/or RNA sequencer.

The system may further comprise at least one microorganism container or at least one hydrogen-containing liquid production enhancer container or a combination thereof.

The at least one microorganism container and/or the at least one hydrogen-containing liquid production enhancer container may be connected to the enclosed bioreactor by an additive solid pathway or an additive liquid pathway.

For the avoidance of doubt, all features relating to the method of the present invention also relate, where appropriate, to the system of the present invention and vice versa.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention will now be more particularly described with reference to the following examples and figures, in which; Figure 1 is a schematic illustration of a system for increasing production of a hydrogen-containing liquid from an enclosed bioreactor according to some embodiments herein; and Figure 2 is a schematic illustration of a system for increasing production of a hydrogen-containing liquid from an enclosed bioreactor according to some embodiments herein.

The foregoing summary, as well as the following detailed description of the embodiments, will be better understood when read in conjunction with the attached drawings. For the purpose of illustration, there are shown in the drawings some embodiments, which may be preferable. It should be understood that the embodiments depicted are not limited to the precise details shown. Unless otherwise noted, the drawings are not to scale.

DETAILED DESCRIPTION

Unless otherwise noted, all measurements are in standard metric units.

Unless otherwise noted, all instances of the words "a," "an," or "the" can refer to one or more than one of the word that they modify.

Unless otherwise noted, the phrase "at least one of" means one or more than one of an object. For example, "at least one of a single walled carbon nanotube, a double walled carbon nanotube, and a triple walled carbon nanotube" means a single walled carbon nanotube, a double walled carbon nanotube, or a triple walled carbon nanotube, or any combination thereof.

Unless otherwise noted, the term "about" refers to ±10% of the non-percentage number that is described, rounded to the nearest whole integer. For example, about 100 mm, would include 90 to 110 mm. Unless otherwise noted, the term "about" refers to ±5% of a percentage number. For example, about 20% would include 15 to 25%. When the term "about" is discussed in terms of a range, then the term refers to the appropriate amount less than the lower limit and more than the upper limit. For example, from about 100 to about 200 mm would include from 90 to 220 mm.

Unless otherwise noted, properties (height, width, length, ratio etc.) as described herein are understood to be averaged measurements.

Unless otherwise noted, the terms "provide", "provided" or "providing" refer to the supply, production, purchase, manufacture, assembly, formation, selection, configuration, conversion, introduction, addition, or incorporation of any element, amount, component, reagent, quantity, measurement, or analysis of any method or system of any embodiment herein.

Unless otherwise noted, the term "non-native" refers to a microorganism that is not naturally occurring in a particular location, such as a particular subterranean formation.

Unless otherwise noted, the term "recombinant microorganism" refers to a microorganism that does not occur in nature and is the synthetic product of genetic manipulation.

Unless otherwise noted, the term "hydrocarbon" refers to a compound that contains only contains hydrogen and carbon atoms.

Unless otherwise noted, the term "gas path" is interchangeable with the term "gas flow path." Unless otherwise noted, the term "gas path" refers to an enclosed solid structure or channel that a gas can move or be pumped through. For example, in various embodiments of the systems and methods disclosed herein, a gas path includes one or more pipes and/or tubes connected to or connected through one or more valves or pumps, so long as gas can flow or be pumped continuously through the structure of the gas path.

Unless otherwise noted, the term "liquid path" is interchangeable with the term "liquid flow path." Unless otherwise noted, the term "liquid path" refers to an enclosed solid structure or channel that a gas can move or be pumped through. For example, in various embodiments of the systems and methods disclosed herein, a gas path includes one or more pipes and/or tubes connected to or connected through one or more valves or pumps, so long as gas can be made to flow or be pumped continuously through the structure of the gas path.

Unless otherwise noted, the term "electrically connected" refers to connecting two or more objects such they can conduct electricity.

Unless otherwise noted, the term "biomass" refers to a product which can contain one or more microorganisms, such as alga (phototrophic organisms), living or dead, colonies of those organisms, and/or the contents of one or more microorganisms, such as enzymes, cytoplasm, nutrients, and the like. An example of a "biomass" can include alga that have been mechanically disrupted.

Unless otherwise noted, the term "enclosed" or "enclosure" refers to a structure that is sealable or resealable, such that when the structure is sealed, the contents of the structure are not free to mix with the open air.

Unless otherwise noted, the term "hydrogen-containing liquid" refers to a molecule that contains hydrogen atoms and from 80% to 100% weight of the compound, relative to the total weight of the compound, is a liquid or liquid slurry at standard temperature and pressure.

EXAMPLES

Example 1: Initial set-up for a depleted oil well.

Purchasing or leasing land having a depleted oilwell with a wellbore and a well casing already in place such that the wellbore and well casing extend into a subterranean formation that has been substantially depleted of hydrocarbons. Attaching a valve assembly to the head of the wellbore such that the valves of the valve assembly can control what enters and leaves the wellbore. A suitable valve assembly can be purchased from oil field service companies such as Mogas, Suez Water Technologies, and Halliburton, among others.

Using a bulldozer to dig a pool into the surface within about 100 to 200 meters out of the valve assembly of the depleted oil well. Digging the pool to a depth up about 5 feet any length and width of about 100 meters. The pool would be filled with water and alga of the genera Chlorella or Scenedesmus which can be purchased from UTEX Culture Collection of Algae at the University of Texas at Austin. A series of rods would be extended over the length and width of the pool to form a support structure, and a transparent polyethylene cover would be used to seal the top of the pool, making it substantially airtight. The covered pool would serve as an algal bioreactor.

A free-standing structure would be connected by one or more gas pipes to the algal reactor to form a hydrogen separation building. The hydrogen separation building would be connected to the subterranean formation either directly by drilling a wellbore into the subterranean formation or by one or more pipes connecting to the valve assembly. The freestanding structure would contain a T-junction connecting a gas path from the subterranean formation to a hydrogen selective membrane, where in on one side of the hydrogen selective membrane (the filtered hydrogen side) the hydrogen separator is connected to a hydrogen storage tank by a gas pipe. A suitable hydrogen selective membrane can be hollow microfiber membranes, which can be purchased from Generon located in California, among other suppliers. Alternatively, palladium-based membranes, such as those available from HySep, can be used for hydrogen separation. The other side of the membrane (the carbon dioxide side) would be connected to the algal bioreactor by a gas pipe.

The valve assembly would be connected to two containers, one serving as a microorganism container and one serving as a hydrogen production enhancer container. The valve assembly would further connect to a genetic material testing facility wherein the genetic material testing facility includes a DNA and/or RNA sequencer and would further connect the valve assembly to the DNA and/or RNA sequencer, such that sequencing could be controlled by a computer or remotely. A suitable DNA and/or RNA sequencer would include the Minion nanopore sequencer, which can be commercially purchased from Oxford Nanopore Technologies located in the United Kingdom.

The algal bioreactor would further be connected to the subterranean formation either directly by a well bore and liquid tube or indirectly by connecting the algal bioreactor to the valve assembly.

Example 2: Increasing hydrogen production from a subterranean formation having a low amount of hydrogen producing microorganisms.

Providing the set up according to Example 1 above, with the following changes.

Taking a gas sample of the hydrogen produced by the subterranean information and analyzing the amount of hydrogen in the gas sample using a gas chromatograph with PDHID (Pulse Discharge Helium Ionization Detection), which can be purchased from Custom Solutions Group which is located in Houston, TX. Determining that the hydrogen output is too low.

Taking a liquid sample from the subterranean formation. Performing a bulk DNA/RNA extraction by performing the steps detailed in the DNeasy PowerSoil Pro Kit from Qiagen (Hilden, Germany). Quantifying the amount of DNA/RNA using real-time PCR and primers that target the 165 rRNA gene. Sequencing the DNA/RNA from the samples using a commercially available kit such as the 16S sequencing kit, which is commercially available from Oxford Nanopore Technologies.

Further testing the liquid sample to determine pH, temperature, and level of nutrients present in the liquid sample.

Analyzing the data from the microorganism population and determining that there are microorganisms present in the subterranean formation, but that less than 1% of the microorganisms present produce hydrogen. Adding 1-50 barrels of -10e cells/nnL of a nonnative hydrogen producing microorganism, such as Clostridium spp., which is known to be a hydrogen producing organism and compatible with a pH of 5-8 and temperature of 25- 35°C (77-95F), until the amount is projected to be over 20% of the total microorganisms present. Suitable Clostridium can be purchased from ATCC, which is located in Manassas, VA.

Harvesting an amount of hydrogen by pumping the gasses from the subterranean formation through the hydrogen selective filter into a hydrogen storage tank at a rate of about 0.3 tons/hr to 30 tons/hr, wherein the percentage of hydrogen in the gas sample is increased by at least 10%. Pumping they non-hydrogen gases such as carbon dioxide into the algal bioreactor. Pumping water, nutrients, and alga from the algal reactor as needed into the subterranean formation to feed the reaction mixture.

Using DNA/RNA sequencing to monitor liquid samples about once a month to ensure that the amount of hydrogen producing microbes does not fall below 20% of the total amount of microbes present.

Example 3: Producing hydrogen carriers in the subterranean formation.

Providing the setup according to Example 1 or Example 2 above, except that the DNA/RNA analysis is used to determine presence of hydrogen carrier producing microorganisms is less than 1%.

Adding 1-50 barrels of -10' cells/mL of a non-native hydrogen carrier producing microorganism, such as a recombinant Methanothermobacter which are known to be hydrogen carrier (methanol) producing organisms until the amount is projected to be over 20% of the total microorganisms present. Suitable anaerobic methanotrophs can be isolated from landfills or anaerobic digesters.

Harvesting an amount of hydrogen carriers by pumping the liquids from the subterranean formation into a hydrogen carrier storage tank at a rate of about 0.3 tons/hr to 30 tons/hr, wherein the percentage of hydrogen carrier in the liquid sample is increased by at least 10%. Pumping they non-hydrogen gases such as carbon dioxide into the algal bioreactor. Pumping water, nutrients, and alga from the algal reactor as needed into the subterranean formation to feed the reaction mixture.

Claims

CLAIMS1 A method of increasing production of a hydrogen-containing liquid from an enclosed bioreactor comprising: providing a baseline reaction mixture in the enclosed bioreactor, wherein the baseline reaction mixture includes a substrate, water, and a baseline amount of at least one microorganism, wherein the substrate includes a nitrogen source, an unsaturated hydrocarbon having from 2 to 120 carbon atoms, methane, hydrogen, or a combination thereof, wherein the hydrogen-containing liquid includes ammonia, ammonium, methanol, a saturated hydrocarbon having from 2 to 120 carbon atoms, or a combination thereof; producing baseline microorganism data on an identity and a baseline percentage of the at least one microorganism, relative to a baseline total percentage of microorganisms in the baseline reaction mixture, by performing DNA and/or RNA sequencing of a baseline microorganism sample from the baseline reaction mixture; measuring a baseline amount of hydrogen-containing liquid in a baseline sample collected from the enclosed bioreactor; increasing production of the hydrogen-containing liquid from the enclosed bioreactor by forming a synthetic reaction mixture, and harvesting the hydrogen-containing liquid from the enclosed bioreactor at a hydrogen-containing liquid harvesting rate by separating the hydrogen-containing liquid from solids and other liquids by transferring the hydrogen-containing liquid into a hydrogen-containing liquid storage container, and forming the synthetic reaction mixture by: adding at least one non-native hydrogen-containing liquid producing microorganism until a percentage of the non-native hydrogen-containing liquid producing microorganism in the synthetic reaction mixture is at least 20% of a total amount of microorganisms in the synthetic reaction mixture; or adding at least one hydrogen-containing liquid production enhancer or a palladium-containing catalyst to the baseline reaction mixture until a post-baseline amount of hydrogen-containing liquid in a post-baseline sample collected from the enclosed bioreactor is at least 10% higher than the baseline amount of hydrogen-containing liquid; or adding at least one recombinant microorganism to the baseline reaction mixture until a percentage of the at least one recombinant microorganism in the synthetic reaction mixture is at least 20% of a total amount of microorganisms in the reaction mixture, or a combination thereof; and wherein the enclosed bioreactor is a bioreactor subterranean formation, a bioreactor landfill enclosure, or a combination thereof.
2. The method of claim 1, wherein the substrate or hydrogen containing liquid contains from 2-70 carbons, or from 2-40 carbon atoms.
3 The method of any of the preceding claims 1 to 2, wherein the nitrogen source includes nitrogen gas, agriculture waste, soy protein isolate, blood meal, feather meal, dried fish, yeast extract, nitrates, nitrites, urea, soy flour, peanut cake, peptone, beef extract, or a combination thereof.
4. The method of any of the preceding claims 1 to 3, further comprising: after providing the baseline reaction mixture, but before forming the synthetic reaction mixture, producing baseline environmental data from the baseline reaction mixture, wherein the baseline environmental data includes one or more measurements of a baseline environmental sample from the baseline reaction mixture, wherein the baseline microorganism sample and the baseline environmental sample can be the same or different.
The method of any of the preceding claims claim 1 to 4, wherein the substrate includes methane, the hydrogen-containing liquid includes methanol, and the at least one non-native hydrogen-containing liquid producing microorganism or the at least one recombinant microorganism has a genus of Methanothermobacter, Methanosaeta, Methanococcales, Candidatus, Desulfobulbus, Desulfocapsa, Desulfofustis, Desuffopila, Desulforhophalus, Desuffotalea, Desuffurivibrio, Methylomirabilis, uncultured anaerobic Methanotrophic archaea (AN ME-1), uncultured ANME-2, uncultured ANME-3, or combination or mixture thereof; or wherein the substrate includes a nitrogen source, the hydrogen-containing liquid includes ammonia or ammonium, and the at least one non-native hydrogen-containing liquid microorganism or the at least one recombinant microorganism has a genus of Methanothermobacter, Methanosaeta, Methanococcales, Candidatus, Desulfobulbus, Desulfocapsa, Desulfofustis, Desuifopila, Desulforhophalus, Desulfotalea, Desulfurivibrio, Methylomirabilis, uncultured anaerobic Methanotrophic archaea (ANME-1), uncultured ANME-2, uncultured ANME-3 or a family of Methanoperedenaceae, Methanobacteriaceae, Methanocaidococcaceae, Desulfobulbaceae, Candidatus Methanoperedenaceae, wherein the at least one non-native hydrogen-containing liquid producing microorganism or the at least one recombinant microorganism can be the same or different.
6. The method of claim 5, wherein the recombinant microorganism expresses at least one Coenzyme M reductase and or dehydrogenase protein having a gene sequences at least 95% identical to SEQ ID NO. [rnmg:MTBMA_c15480], [mth:MTH_1015], [mmg:MTBMA_c15520], [mmg:MTBMA_c15490], [mth:MTH_1166], [mth:MTH_1167], [eco:b4346], [eco:134345], [ag:AAA22593], [mea:Mex_1p4538, [mea:Mex_1p4535], [ag:ACS29499], [ag:CAH55641], [mrd:Mrad2831_0508], by expressing a non-native Coenzyme M reductase and or dehydrogenase expressing nucleotide sequence, wherein an amount of hydrogen-containing liquid produced or protein produced by the recombinant microorganism is greater than that produced relative to a control microorganism lacking the nonnative Coenzyme M reductase and or dehydrogenase expressing nucleotide sequences.
7 The method of any of the preceding claims 1 to 6, wherein the substrate includes the saturated hydrocarbon having from 2 to 40 carbon atoms, the hydrogen-containing liquid includes the saturated hydrocarbon having from 2 to 40 carbon atoms, and the palladium-containing catalyst includes palladium on carbon, palladium on barium carbonate, palladium on barium sulfate, palladium on calcium carbonate, palladium on silica gel, Pd(OH)2, or a combination or a mixture thereof.
8. The method of any of the preceding claims 1 to 7, wherein the hydrogen-containing liquid harvesting rate is from about 0.1 Uhr to about 106 Uhr; and/or wherein the unsaturated hydrocarbon having from 2 to about 40 carbon atoms includes an alkene, an alkyne, an aromatic hydrocarbon, or a polyaromatic hydrocarbon; and/or wherein the bioreactor subterranean formation includes a natural formation, non-natural formation, a hydrocarbon-bearing formation, a natural gas-bearing formation, a methane-bearing formation, a depleted hydrocarbon formation, a depleted natural gas-bearing formation, a wellbore, or a combination thereof; and/or wherein the bioreactor landfill enclosure includes a landfill that is enclosed by a building material, wherein the building material includes at least one of a brick, a cement, a plastic, a non-natural rubber, a geomembrane of any kind, concrete, steel, a glass, or a combination thereof.
9. The method of any of the preceding claims 1 to 8, wherein the hydrogen-containing liquid production enhancer is a biocidal inhibitor, a methanogenesis inhibitor, a sulfate reduction inhibitor, a nitrate reduction inhibitor, an iron reduction inhibitor, or a combination thereof, and wherein the biocidal inhibitor is glutaraldehyde, a quaternary ammonium compound, formaldehyde, a formaldehyde releaser such as 3,3'-methylenebis[5-methyloxazolidine], dibromonitrilopropionamide, tetra kis hydroxymethyl phosphonium sulfate, chlorine dioxide, peracetic acid, tributyl tetradecyl phosphonium chloride, methylisothiazolinone, chloromethylisothiazolinone, sodium hypochlorite, dazomet, dimethyloxazolidine, trimethyloxazolidine, N-bromosuccinimide, bronopol, or 2-propenal, or a mixture thereof, and/or wherein the methanogenesis inhibitor is bromethane sulfonic acid, an aminobenzoic acid, 2-bromoethanesulfonate, 2-chloroethanesulfonate, 2-mercaptoethanesulfonate, lumazine, a fluoroacetate, nitroethane, or 2-n itropropanol, or a mixture thereof, and/or wherein the sulfate reduction inhibitor is a molybdate salt, a nitrate salt, a nitrite salt, a chlorate salt, or a perch lorate salt or a mixture thereof, and/or wherein the nitrate reduction inhibitor is sodium chlorate, a chlorate salt, or a perchlorate salt, or a mixture thereof.
10. The method of any of the preceding claims 1 to 9, wherein forming the synthetic reaction mixture includes: adding the at least one non-native hydrogen-containing liquid producing microorganism until a percentage of the non-native hydrogen-containing liquid producing microorganism in the synthetic reaction mixture is at least 51% of a total amount of microorganisms in the synthetic reaction mixture; or adding the at least one hydrogen-containing liquid production enhancer or the palladium-containing catalyst to the baseline reaction mixture until a post-baseline amount of hydrogen-containing liquid in a post-baseline gas sample of liquids collected from the enclosed bioreactor is at least 51% higher than the baseline amount of hydrogen-containing liquid; or adding the at least one recombinant microorganism to the baseline reaction mixture until a percentage of the at least one recombinant microorganism in the synthetic reaction mixture is at least 51% of a total amount of microorganisms in the reaction mixture.
11. The method of any of the preceding claims 1 to 10, further comprising: harvesting hydrogen from the enclosed bioreactor at a hydrogen harvesting rate, and separating the hydrogen from other gasses by filtering the hydrogen through a hydrogen-selective membrane filter and transferring the hydrogen into a hydrogen storage container.
12.A system for increasing production of a hydrogen-containing liquid from an enclosed bioreactor comprising: the enclosed bioreactor, a hydrogen-containing liquid storage container, and a hydrogen-containing liquid separator, wherein the reaction mixture includes a substrate, water, and a baseline amount of at least one microorganism, wherein the hydrogen-containing liquid separator includes at least one hydrogen-containing liquid membrane filter, at least one solids filter, or at least one distillation apparatus; wherein the enclosed bioreactor is connected to the hydrogen-containing liquid separator by a hydrogen-containing liquid path, wherein the hydrogen-containing liquid separator is connected to the hydrogen-containing liquid container by a purified hydrogen-containing liquid path.
13. The system of claim 12, wherein the enclosed bioreactor has a volume of from about 100 m3 to about 4 x 109 rn3, and/or wherein the hydrogen-containing liquid storage container includes a liquid tank or a hydrogen-containing liquid surface formation.
14. The system of any of preceding claims 12 to 13, further comprising: a genetic material testing facility within about 1000 meters of a resealable opening of the enclosed bioreactor and connected to the resealable opening by a genetic material testing liquid pathway, wherein the genetic material testing facility contains at least one DNA and/or RNA sequencer.
15. The system of any of preceding claims claim 12 to 14, further comprising: at least one microorganism container or at least one hydrogen-containing liquid production enhancer container or a combination thereof, wherein the at least one microorganism container or the at least one hydrogen-containing liquid production enhancer container are connected to the enclosed bioreactor by an additive solid pathway or an additive liquid pathway.