IP Library › Granted Patent US 12,595,494
Granted Patent B2
US 12,595,494 · App. 18/379,079 · Granted Apr 7, 2026

Biological production of multi-carbon compounds from methane

Inventors: William J. Coleman (Redwood City, CA); Genevieve M. Vidanes (San Francisco, CA); Guillaume Cottarel (Mountain View, CA); Sheela Muley (Fremont, CA); Roy Kamimura (Daly City, CA); Akbar F. Javan (Chapel Hill, NC); Jianping Sun (Belmont, CA); Eli S. Groban (San Francisco, CA)
Assignee: BioVerde Tech LLC
C12P7/16C12N9/0006C12N9/1022C12N9/1025C12N9/88C12N15/52C12Y101/01001C12Y101/01004C12Y101/01086C12Y202/01006C12Y203/03006C12Y401/01072C12Y402/01009C12Y402/01033C12Y403/01019Y02E50/10
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Quick Facts
Patent No.
US 12,595,494
App. No.
18/379,079
Granted
Apr 7, 2026
Kind
B2
Abstract

Multi-carbon compounds such as ethanol, n-butanol, sec-butanol, isobutanol, tert-butanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters, 2,3-butanediol and the like, are important industrial commodity chemicals with a variety of applications. The present invention provides metabolically engineered host microorganisms which metabolize methane (CH 4 ) as their sole carbon source to produce multi-carbon compounds for use in fuels (e.g., bio-fuel, bio-diesel) and bio-based chemicals. Furthermore, use of the metabolically engineered host microorganisms of the invention (which utilize methane as the sole carbon source) mitigate current industry practices and methods of producing multi-carbon compounds from petroleum or petroleum-derived feedstocks, and ameliorate much of the ongoing depletion of arable food source “farmland” currently being diverted to grow bio-fuel feedstocks, and as such, improve the environmental footprint of future bio-fuel, bio-diesel and bio-based chemical compositions.

Claims (22)

1 . A genetically modified methanotroph comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the acetolactate synthase can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, and wherein said genetically modified methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway.

2 . The genetically modified methanotroph of claim 1 , wherein the genetically modified methanotroph further comprises a heterologous polynucleotide encoding a ketoacid decarboxylase (KDC), wherein the ketoacid decarboxylase can catalyze the conversion of ketoisovalerate to isobutryaldehyde.

3 . The genetically modified methanotroph of claim 2 , wherein the genetically modified methanotroph further comprises a heterologous polynucleotide encoding a ketol-acid reductoisomerase (KARI), a heterologous polynucleotide encoding a dihydroxy-acid dehydratase (DHAD), and a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH); wherein the ketol-acid reductoisomerase can catalyze the conversion of acetolactate to 2,3-dihydroxyisovalerate; wherein the dihydroxy-acid dehydratase can catalyze the conversion of 2,3-dihydroxyisovalerate to ketoisovalerate; and wherein the alcohol dehydrogenase can catalyze the conversion of isobutyraldehyde to isobutanol.

4 . The genetically modified methanotroph of claim 3 , wherein the genetically modified methanotroph further comprises a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH), wherein the ketoacid decarboxylase can catalyze the conversion of isobutyraldehyde to isobutanol.

5 . The genetically modified methanotroph of claim 4 , wherein the ketoacid decarboxylase (KDC) comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 8.

6 . The genetically modified methanotroph of claim 1 , wherein the genetically modified methanotroph further comprises a heterologous polynucleotide encoding a ketol-acid reductoisomerase (KARI), heterologous polynucleotide encoding a dihydroxy-acid dehydratase (DHAD), and a heterologous polynucleotide encoding a ketoacid decarboxylase (KDC), and a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH); wherein the acetolactate synthase can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2; wherein the ketol-acid reductoisomerase can catalyze the conversion of acetolactate to 2,3-dihydroxyisovalerate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 4; wherein the dihydroxy-acid dehydratase can catalyze the conversion of 2,3-dihydroxyisovalerate to ketoisovalerate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 6; wherein the ketoacid decarboxylase can catalyze the conversion of ketoisovalerate to isobutryaldehyde and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 8; and wherein the alcohol dehydrogenase can catalyze the conversion of isobutyraldehyde to isobutanol and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 10.

7 . The genetically modified methanotroph of claim 6 , wherein the genetically modified methanotroph further comprises a polynucleotide sequence encoding for the alcohol dehydrogenase (ADH) and a promoter, wherein said promoter can direct the expression of the alcohol dehydrogenase in the genetically modified methanotroph.

8 . The genetically modified methanotroph of claim 2 , wherein the genetically modified methanotroph further comprises a polynucleotide sequence encoding for the ketoacid decarboxylase (KDC), the alcohol dehydrogenase (ADH), and a promoter, wherein said promoter can direct the expression of the ketoacid decarboxylase and the alcohol dehydrogenase (ADH) in the genetically modified methanotroph.

9 . The genetically modified methanotroph of claim 3 , wherein the genetically modified methanotroph further comprises a polynucleotide sequence encoding for the acetolactate synthase (ALS), the ketol-acid reductoisomerase (KARI), the dihydroxy-acid dehydratase (DHAD), the ketoacid decarboxylase (KDC), the alcohol dehydrogenase (ADH) and a promoter, wherein said promoter can direct the expression of the acetolactate synthase (ALS), the ketol-acid reductoisomerase (KARI), the dihydroxy-acid dehydratase (DHAD), the ketoacid decarboxylase (KDC), and the alcohol dehydrogenase (ADH) in the genetically modified methanotroph.

10 . The genetically modified methanotroph of claim 9 , wherein said promoter is constitutive.

11 . The genetically modified methanotroph of claim 9 , wherein said promoter is inducible.

12 . The genetically modified methanotroph of claim 1 , wherein said genetically modified methanotroph is from the genus Methylobacter, Methylomicrobium, Methylomonas, Methylocaldum, Methylococcus, Methylosoma, Methylosarcina, Methylothermus, Methylohalobius, Methylogaea, Methylovulum, Crenothrix, Clonothrix, Methylosphaera, Methylocapsa, Methylocella, Methylosinus, Methylocystis , or Methyloacidophilum.

13 . The genetically modified methanotroph of claim 1 , wherein said genetically modified methanotroph is from the genus Methylococcus.

14 . The genetically modified methanotroph of claim 13 , wherein said genetically modified methanotroph is from the species Methylococcus capsulatus.

15 . The genetically modified methanotroph of claim 13 , wherein said genetically modified methanotroph is from the strain Methylococcus capsulatus strain Bath.

16 . A method of making a multi-carbon compound comprising:

(a) contacting a genetically modified methanotroph with a multi-carbon product precursor comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the ALS can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, and wherein said genetically modified methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway; and

(b) growing said genetically modified methanotroph in conditions to produce a multi-carbon compound.

17 . The method of claim 16 , wherein said multi-carbon product precursor is methane.

18 . The method of claim 16 wherein said multi-carbon compound is isobutanol.

19 . The method of claim 16 wherein said multi-carbon compound is 1-butanol.

20 . A genetically modified methanotroph capable of converting methane to a multi-carbon product comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the ALS can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, and wherein said genetically modified methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway.

Assignments (5)
CERTIFICATE OF RESTATEMENT Recorded Nov 24, 2025
From: INTREXON CORPORATION
To: PRECIGEN, INC.
Reel/Frame 073770/0389 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 10, 2025
From: COLEMAN, WILLIAM; VIDANES, GENEVIEVE; COTTAREL, GUILLAUME; MULEY, SHEELA; KAMIMURA, ROY; JAVAN, ABKAR; SUN, JIANPING; GROBAN, ELI S.
To: INTREXON CORPORATION
Reel/Frame 072533/0364 →
CHANGE OF NAME Recorded Oct 10, 2025
From: INTREXON CORPORATION
To: PRECIGEN, INC.
Reel/Frame 073839/0276 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 2, 2025
From: PRECIGEN, INC.
To: BIOVERDE TECH LLC
Reel/Frame 072446/0769 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 15, 2025
From: PRECIGEN, INC.
To: BIOVERDE TECH LLC
Reel/Frame 072033/0001 →
Continuity (7)
Continuation 17103516 · Nov 24, 2020
Continuation 15648920 · Jul 13, 2017
Continuation 15192290 · Jun 24, 2016
Division 14989859 · Jan 7, 2016
Division 14206835 · Mar 12, 2014
Provisional Application 61782830 · Mar 14, 2013
Related Publication 20240368638A1 · Nov 7, 2024
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