# US 12,110,529 B2 · Methods and systems using ionophores to control contamination in fermentation of gaseous substrates

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,110,529 B2
- Document ID: US12110529B2
- Status: Granted
- Kind: B2
- Application number: 16/471,453
- Filed: 2017-12-14
- Published: 2021-05-06
- Granted: 2024-10-08
- Earliest claimed: 2016-12-22
- Assignees: Synata Bio, Inc.
- Applicants: Synata Bio, Inc.
- Inventors: Jennine B. Terrill
- Web page: https://patentgrep.com/patent/US12110529B2

## CPC

- C12P 7/065: {with microorganisms other than yeasts}
- C12M 21/12: {for producing fuels or solvents (C12M21/04 takes precedence; liquid carbonaceous fuels C10L1/00, solid fuels C10L5/00)}
- C12M 43/02: {Bioreactors or fermenters combined with devices for liquid fuel extraction; Biorefineries}
- C12P 7/04: acyclic
- C12P 7/14: Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
- C12P 7/16: Butanols
- C12P 7/40: containing a carboxyl group {including Peroxycarboxylic acids}
- C12P 7/54: Acetic acid (vinegar C12J)
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

Provided is a method of anaerobically fermenting a gaseous substrate to form a liquid product, the method comprising: (a) introducing the gaseous substrate into a bio-reactor, the gaseous substrate comprising at least one of the following constituents: carbon monoxide, carbon dioxide, and hydrogen, (b) the bio-reactor comprising a fermentation broth therein, the fermentation broth containing at least two types of microorganisms, one type comprising at least one fermenting species, and the other type comprising at least one competing species; (c) introducing at least one type of ionophore into the reactor, the ionophore having selectivity for preferentially inhibiting the at least one competing species from growing and/or producing an undesired product; and (d) allowing the gaseous substrate to ferment by exposure to the at least one fermenting species, to produce the liquid product and a system for doing the same.

## Claims

### Claim 1

A method of anaerobically fermenting a syngas to form a liquid product comprising an alcohol or acid, the method comprising:
a. introducing the syngas into a bio-reactor, wherein the syngas comprises carbon monoxide, carbon dioxide, and hydrogen,
i. the bio-reactor comprising a fermentation broth therein, the fermentation broth containing at least two types of microorganisms, one type comprising at least one fermenting microorganism species, and the other type comprising at least one competing microorganism species;
b. introducing at least one type of ionophore into the bio-reactor, the ionophore having selectivity for preferentially inhibiting the at least one competing microorganism species from growing and/or producing an undesired product; and
c. allowing the syngas to ferment by exposure to the at least one fermenting microorganism species, to produce the liquid product; wherein:
the fermenting species is Clostridium autoethanogenum; and
the competing species is Acetobacterium woodii.

### Claim 2 (depends on 1)

The method of claim 1, wherein the competing species is a contaminant and the one fermenting species is a C1 fixing microorganism.

### Claim 3 (depends on 2)

The method of claim 2, further comprising introducing media into the bio-reactor.

### Claim 4 (depends on 1)

The method of claim 1, wherein the liquid product is one or more of ethanol, n-propanol, iso-propanol, n-butanol, iso-butanol, n-pentanol, n-hexanol, acetic acid, propanoic acid, butanoic acid, pentatonic acid, and hexanoic acid.

### Claim 5 (depends on 1)

The method of claim 1, wherein the competing species uses sodium gradients for adenosine triphosphate (ATP) production, and wherein the ionophore inhibits the sodium gradients from producing ATP.

### Claim 6 (depends on 1)

The method of claim 1, wherein the fermenting species uses hydrogen gradients for ATP production.

### Claim 7 (depends on 1)

The method of claim 1, wherein the ionophore is one or more of the following: 12-crown-4, 15-crown-6, 18-crown-6, dibenzo-18-crown-6, and diaza-18-crown-6, A23187, 4-bromo-A23187, alamethicin, beauvericin, bapta(AM), calcimycin, cezomycin, carbonyl cyanide m-chlorophenyl hydrazone, CA 1001, enniatin, gramicidin A, hellebrin, ionomycin, lasalocid, monensin, nigericin, nonactin, phenamil methanesulfonate, salinomycin, tetonasin, ETH2120, ETH 227, ETH 157, 2,3:11,12-Didecalino-16-crown-5, DD-16-C-5, ETH 4120, Dodecylmethylmalonic acid bis[(12-crown-4)methyl ester], Bis[(12-crown-4)methyl] 2,2-didodecylmalonate, 4-tert-Butylcalix[4]arene-tetraacetic acid tetraethyl ester, BME 44 and valinomycin.

### Claim 8

A fermentation system for anaerobically fermenting a syngas to form a liquid product, the system comprising:
a. a fermentation vessel;
b. a syngas supply conduit for introducing the syngas into the fermentation vessel;
c. an aqueous fermentation broth disposed in the fermentation vessel, the fermentation broth containing at least two types of microorganisms, one type comprising at least one fermenting species, and the other type comprising at least one competing species;
d. an ionophore supply conduit in communication with the fermentation vessel for introducing at least one type of ionophore into the vessel, the ionophore having selectivity for preferentially inhibiting the at least one competing species from growing and/or producing an undesired product; and
e. a liquid recovery conduit for collecting the liquid product after the syngas is allowed to ferment by exposure to the at least one fermenting species;
the fermenting species is Clostridium autoethanogenum; and
the competing species is Acetobacterium woodii.

### Claim 9 (depends on 8)

The system of claim 8, wherein the liquid product is at least one alcohol, at least one acid, or any combination thereof.

### Claim 10 (depends on 8)

The system of claim 8, wherein the liquid product is one or more of ethanol, n-propanol, iso-propanol, n-butanol, iso-butanol, n-pentanol, n-hexanol, acetic acid, propanoic acid, butanoic acid, pentatonic acid, and hexanoic acid.

### Claim 11 (depends on 8)

The system of claim 8, wherein the pH is from about 3 to about 7.

### Claim 12 (depends on 8)

The system of claim 8, wherein the fermenting species uses hydrogen gradients for ATP production.

### Claim 13 (depends on 8)

The system of claim 8, wherein the ionophore is one or more of the following: 12-crown-4, 15-crown-6, 18-crown-6, dibenzo-18-crown-6, and diaza-18-crown-6, A23187, 4-bromo-A23187, alamethicin, beauvericin, bapta(AM), calcimycin, cezomycin, carbonyl cyanide m-chlorophenyl hydrazone, CA 1001, enniatin, gramicidin A, hellebrin, ionomycin, lasalocid, monensin, nigericin, nonactin, phenamil methanesulfonate, salinomycin, tetonasin, ETH2120, ETH 227, ETH 157, 2,3:11, 12-Didecalino-16-crown-5, DD-16-C-5, ETH 4120, Dodecylmethylmalonic acid bis[(12-crown-4)methyl ester], Bis[(12-crown-4)methyl] 2,2-didodecylmalonate, 4-tert-Butylcalix[4]arene-tetraacetic acid tetraethyl ester, BME 44 and valinomycin.

### Claim 14 (depends on 8)

The system of claim 8, wherein at least a portion of the syngas is prepared from coal.

### Claim 15 (depends on 8)

The system of claim 8, wherein the syngas contains at least one of the following: water vapor, nitrogen, methane, ammonia, and hydrogen sulfide.

### Claim 16 (depends on 1)

The method of claim 1, wherein at least a portion of the syngas is prepared from coal.

### Claim 17 (depends on 1)

The method of claim 1, wherein the syngas contains at least one of the following: water vapor, nitrogen, methane, ammonia, and hydrogen sulfide.

### Claim 18 (depends on 7)

The method of claim 7, wherein the ionophore is monensin.

### Claim 19 (depends on 13)

The system of claim 13, wherein the ionophore is monensin.