IP Library › Granted Patent US 12,486,513
Granted Patent B2
US 12,486,513 · App. 16/728,509 · Granted Dec 2, 2025

Modulation of carbon flux through the meg and C3 pathways for the improved production of monoethylene glycol and C3 compounds

Inventors: Ane Fernanda Beraldi Zeidler (Campinas, BR); Beatriz Leite Magalhaes (Campinas, BR); Lucas Pedersen Parizzi (Campinas, BR); Veronica Maria Rodege Gogola Kolling (Campinas, BR)
Assignee: Braskem S.A.
C12N15/52C12P7/28C12Y401/01047C12Y402/03003
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,486,513
App. No.
16/728,509
Granted
Dec 2, 2025
Kind
B2
Abstract

The present disclosure provides methods of modulating the flux of carbon through the monoethylene glycol (MEG) biosynthesis pathway and one or more C3 compound biosynthesis pathways by expressing enzymes that are essential for improving C3 compounds and modulating other genetic aspects of MEG and C3 compound biosynthesis. The disclosure is further drawn to modified microbes comprising the disrupted sequences and overexpressed sequences, and compositions thereof.

Claims (27)

1 . A recombinant Escherichia coli capable of coproducing MEG and one or more C3 compounds, wherein the recombinant E. coli comprises:

a disruption of endogenous E. coli pta and one or more of the following (i) to (iii):

(i) a disruption of endogenous E. coli ackA,

(ii) a disruption of endogenous E. coli arcA, and

(iii) a disruption of endogenous E. coli pka;

wherein the recombinant E. coli comprises a pathway for MEG production; and

wherein one of the one or more C3 compounds is acetone and the recombinant E. coli comprises a pathway for acetone production.

2 . The recombinant E. coli of claim 1 , wherein the recombinant E. coli further comprises one or more of the following:

(i) one or more polynucleotide sequences encoding acetoacetyl COA synthase for converting acetyl-CoA and malonyl-CoA to acetoacetyl-CoA, wherein the acetoacetyl CoA synthase is Streptomyces sp. NphT7, and/or

(ii) one or more polynucleotide sequences encoding (a) hydroxymethylglutaryl-CoA synthase for converting acetyl-CoA and acetoacetyl-CoA to(S)-3-hydroxy-3-methylglutaryl-CoA and (b) hydroxymethylglutaryl-CoA lyase for converting the(S)-3-hydroxy-3-methylglutaryl-CoA acetyl-CoA and acetoacetate, wherein the hydroxymethylglutaryl-CoA synthase is ERG13 and the hydroxymethylglutaryl-CoA lyase is YngG.

3 . The recombinant E. coli of claim 1 , wherein the MEG exhibits an increased yield or titer.

4 . The recombinant E. coli of claim 3 , wherein the increased yield or titer is an increase of at least 2%.

5 . The recombinant E. coli of claim 3 , wherein the increased yield or titer is an increase of at least 15%.

6 . The recombinant E. coli of claim 1 , wherein the acetone exhibits an increased yield or titer.

7 . The recombinant E. coli of claim 6 , wherein the increased yield or titer is an increase of at least 2%.

8 . The recombinant E. coli of claim 6 , wherein the increased yield or titer is an increase of at least 15%.

9 . The recombinant E. coli of claim 1 , wherein the E. coli utilizes xylose, cellobiose, arabinose, mannose, and/or glucose in the coproduction of the MEG and the one or more C3 compounds.

10 . The recombinant E. coli of claim 1 , wherein the C3 compounds further comprise isopropanol and/or propene.

11 . The recombinant E. coli of claim 1 , wherein the recombinant E. coli further comprises a disruption of endogenous E. coli poxB.

12 . The recombinant E. coli of claim 1 , wherein the recombinant E. coli further comprises one or more of the following:

(i) one or more overexpressed endogenous polynucleotide sequences encoding a CobB regulator, and/or

(ii) one or more overexpressed endogenous polynucleotide sequences encoding an acetyl-CoA synthetase, wherein the acetyl-CoA synthetase is E. coli acs.

13 . The recombinant E. coli of claim 1 , wherein the recombinant E. coli further comprises one or more of the following:

(i) one or more polynucleotide sequences encoding one or more polypeptides comprising means for converting acetyl-CoA and malonyl-CoA to acetoacetyl-CoA, and/or

(ii) one or more polynucleotide sequences encoding one or more polypeptides comprising means for converting acetyl-CoA and acetoacetyl-CoA to (S)-3-hydroxy-3-methylglutaryl-CoA and means for converting the (S)-3-hydroxy-3-methylglutaryl-CoA to acetyl-CoA and acetoacetate.

14 . The recombinant E. coli of claim 13 , wherein the means for converting acetyl-CoA and malonyl-CoA to acetoacetyl-CoA is NphT7.

15 . The recombinant E. coli of claim 13 , wherein the means for converting acetyl-CoA and acetoacetyl-CoA to (S)-3-hydroxy-3-methylglutaryl-CoA is ERG13 and the means for converting the(S)-3-hydroxy-3-methylglutaryl-CoA to acetyl-CoA and acetoacetate is YngG.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 22, 2023
From: ZEIDLER, ANE FERNANDA BERALDI; PARIZZI, LUCAS PEDERSEN; KOLLING, VERONICA MARIA RODEGE GOGOLA
To: BRASKEM S.A.
Reel/Frame 063065/0283 →
Continuity (6)
Provisional Application 62786294 · Dec 28, 2018
Provisional Application 62786298 · Dec 28, 2018
Provisional Application 62786282 · Dec 28, 2018
Provisional Application 62786283 · Dec 28, 2018
Provisional Application 62786304 · Dec 28, 2018
Related Publication 20200208160A1 · Jul 2, 2020
References Cited (65)
US 7531337B2 · Escalante-Semerena et al. · 2009 [cited by applicant]
US 7935511B2 · San et al. · 2011 [cited by applicant]
US 8460726B2 · Harel et al. · 2013 [cited by applicant]
US 8524472B2 · Muramatsu et al. · 2013 [cited by applicant]
US 8637286B2 · Burgard et al. · 2014 [cited by applicant]
US 20050170482A1 · San et al. · 2005 [cited by applicant]
US 20060073577A1 · Ka-Yiu et al. · 2006 [cited by applicant]
US 20070249018A1 · Vemuri et al. · 2007 [cited by applicant]
US 20090253164A1 · Unrean et al. · 2009 [cited by applicant]
US 20150329877A1 · Bockrath · 2015 [cited by applicant]
US 20160265005A1 · Zelle et al. · 2016 [cited by applicant]
US 20160326553A1 · Burgard et al. · 2016 [cited by applicant]
US 20170260551A1 · Koch et al. · 2017 [cited by applicant]
US 20180023101A1 · Koch et al. · 2018 [cited by applicant]
US 20180087074A1 · Reed et al. · 2018 [cited by applicant]
US 20180179558A1 · Koch et al. · 2018 [cited by applicant]
US 20180312887A1 · Lynch et al. · 2018 [cited by applicant]
EP 1546304B1 · 2013 [cited by applicant]
KR 101351879B1 · 2014 [cited by applicant]
WO WO2005087940A1 · 2005 [cited by applicant]
WO 2006034156A2 · 2006 [cited by applicant]
WO 2010076324A1 · 2010 [cited by applicant]
WO WO2011012697A2 · 2011 [cited by applicant]
WO WO2013007786A1 · 2013 [cited by applicant]
WO WO2013126721A1 · 2013 [cited by applicant]
WO WO2013163230A2 · 2013 [cited by applicant]
WO WO2014076232A2 · 2014 [cited by applicant]
WO WO2014102180A1 · 2014 [cited by applicant]
WO WO2015042588A1 · 2015 [cited by applicant]
WO 2017165397A1 · 2017 [cited by applicant]
WO WO2017156166A1 · 2017 [cited by applicant]
WO WO2018013701A1 · 2018 [cited by applicant]
WO WO2018071563A1 · 2018 [cited by applicant]
WO WO2018148703A1 · 2018 [cited by applicant]
Devos et al., (Proteins: Structure, Function and Genetics, 2000, vol. 41: 98-107. [cited by examiner]
Whisstock et al., (Quarterly Reviews of Biophysics 2003, vol. 36 (3): 307-340. [cited by examiner]
Witkowski et al., (Biochemistry 38:11643-11650, 1999. [cited by examiner]
Kisselev L., (Structure, 2002, vol. 10: 8-9. [cited by examiner]
Alkim, Ceren, et al. “Optimization of ethylene glycol production from (D)-xylose via a synthetic pathway implemented in [cited by applicant]
Berzin et al., “Selective production of acetone during continuoussynthesis gas fermentation by engineered biocatalyst [cited by applicant]
Boonstra et al., “The udhA Gene of [cited by applicant]
Cabulong et al., “Enhanced yield of ethylene glycol production from D-xylose bypathway optimization in [cited by applicant]
Canonaco et al., “Metabolic flux response to phosphoglucose isomerase knock-out in [cited by applicant]
Castaño-Cerezo et al., “An insight into the role of phosphotransacetylase (pta) and the acetate/acetyl-CoA node in [cited by applicant]
Causey et al., “Engineering [cited by applicant]
Charusanti et al., “Genetic Basis of Growth Adaptation of [cited by applicant]
Chen, Zhen, et al. “Metabolic engineering of Corynebacterium glutamicum for the de novo production of ethylene glycol from glucose.” Metabolic Engineering (2016); 33: 12-18. [cited by applicant]
Dittrich et al., “Redistribution of Metabolic Fluxes in the Central Aerobic Metabolic Pathway of [cited by applicant]
Ehrenberger et al., “Structure-Guided Engineering of Xylitol Dehydrogenase Cosubstrate Specificity,” Structure, Mar. 2006, 14: 567-575. [cited by applicant]
Hao and Berry, “A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents,” Protein Engineering, Design & Selection, 2004, vol. 17, No.… [cited by applicant]
Jarboe, LR, “YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals,” Applied Microbiology and Biotechnology 2010, 89:249-257. [cited by applicant]
Klasson et al., “Bioconversion of synthesis gas into liquid or gaseous fuels,” Enzyme and Microbial Technology, Aug. 1992, vol. 14, pp. 602-608. [cited by applicant]
Klasson et al., “Bioreactor design for synthesis gas fermentations,” Fuel, (1991), 70:605-614. [cited by applicant]
Klasson et al., “Bioreactors for synthesis gas fermentations,” Resources, Conservation and Recycling, 1991, 5:145-165. [cited by applicant]
Li et al., “Enhanced activity of yqhD oxidoreductase in synthesis of 1,3-propanediol by error-prone PCR,” Progress in Natural Science, 2008, 18: 1519-1524. [cited by applicant]
Lin et al., “Acetyl-CoA synthetase overexpression in [cited by applicant]
Liu et al., “Biosynthesis of ethylene glycol in [cited by applicant]
Patel et al., “Engineering of the catalytic site of xylose isomerase to enhance bioconversion of a non-preferential substrate,” Protein Engineering, Design & Selection, 2012, vol. 25, No. 7, pp. 331-336. [cited by applicant]
Peebo et al., “Coordinated activation of PTA-ACS and TCA cycles strongly reduces overflow metabolism of acetate in [cited by applicant]
Sauer et al., “The Soluble and Membrane-bound Transhydrogenases UdhA and PntAB have Divergent Functions in NADPH Metabolism of [cited by applicant]
Sulzenbacher et al., “Crystal Structure of [cited by applicant]
Vega et al., “Design of Bioreactors for Coal Synthesis Gas Fermentations,” Resources, Conservation and Recycling, (1990), 3:149-160. [cited by applicant]
Vega et al., “Study of gaseous substrate fermentations: Carbon monoxide conversion to acetate. 1. Batch culture,” Biotech. Bioeng., (1989), 34(6):774-784. [cited by applicant]
Vega et al., “Study of Gaseous Substrate Fermentation: Carbon Monoxide Conversion to Acetate. 2. Continuous Culture,” Biotech. Bioeng., (1989), 34(6):785-793. [cited by applicant]
Yomano et al., “Deletion of methylglyoxal synthase gene (mgsA) increased sugar co-metabolism in ethanol-producing [cited by applicant]