Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Engineering microbes and metabolic pathways for the production of ethylene glycol

a technology of metabolic engineering and ethylene glycol, which is applied in the direction of lyases, racemaces/epimerases, transferases, etc., can solve the problems of significant waste, achieve the effect of increasing the amount of 2-phosphoglycerate, increasing the amount of 3-phosphoglycerate, and reducing the flux to 2-phosphoglycera

Inactive Publication Date: 2013-11-28
MASSACHUSETTS INST OF TECH
View PDF5 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent describes a method for increasing the amount of a compound called 3-phosphoglycerate in a cell by reducing the activity of certain enzymes that produce 2-phosphoglycerate. This increase can lead to improved performance and efficiency in cells that require the compound for their metabolism. The patent identifies specific genes that encode these enzymes and proposes techniques for reducing their expression or activity in order to increase 3-phosphoglycerate levels. Overall, the patent presents a way to optimize cellular metabolism and improve cell performance through genetic modification.

Problems solved by technology

Additionally, the production of biodiesel results in significant amounts of waste glycerol, a sugar alcohol which can be used as a substrate.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Engineering microbes and metabolic pathways for the production of ethylene glycol
  • Engineering microbes and metabolic pathways for the production of ethylene glycol
  • Engineering microbes and metabolic pathways for the production of ethylene glycol

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0149]Researching E. coli for the biological production of ethylene glycol (EG) from sugars, we noted the pathway for the degradation of D-arabinose in K-12 strains. In the pathway, the pentose intermediate D-ribulose-1-phosphate is cleaved to yield dihydroxyacetone phosphate (DHAP), which enters into glycolysis, and glycolaldehyde. Glycolaldehyde is typically oxidized to glycolate by aldehyde dehydrogenase A (encoded by the aldA gene), but as in the analogous reaction of L-lactaldehyde to L-1,2-propanediol, glycolaldehyde can also be reduced by the oxidoreductase encoded by fucO. The fluxes through these reactions depend on the availability of oxygen, so we investigated the degradation of D-arabinose under aerobic and anaerobic conditions.

[0150]Cultures of E. coli K-12 MG1655 DE3 ΔendA ΔrecA (referred to as wild-type or WT) and E. coli K-12 MG1655 DE3 ΔendA ΔrecA ΔaldA (referred to as ΔaldA) were grown aerobically and anaerobically on minimal medium with 4 g / L D-arabinose as the ca...

example 2

[0151]Though we have shown that E. coli is capable of generating ethylene glycol from D-arabinose, biomass-derived sugars, such as D-xylose and D-glucose, are much more abundant and therefore are preferred substrates. Because D-xylose is a pentose, we next pursued the utilization of D-xylose for EG production through the previously established D-arabinose degradation pathway. D-tagatose 3-epimerase (DTE) from Pseudomonas cichorii is a promiscuous enzyme that has been shown to interconvert D-xylulose and D-ribulose [Izumori 1993], intermediates of the D-xylose degradation and D-arabinose degradation pathways, respectively. Therefore, DTE (encoded by the gene here referred to as dte) can provide a path by which D-xylose can yield EG, however, conversion of D-xylulose to D-xylulose-5-phosphate, catalyzed by xylulokinase (encoded by xylB), is a competing reaction. We generated E. coli K-12 MG1655 DE3 ΔendA ΔrecA ΔaldA ΔxylB (referred to as ΔaldA ΔxylB) and transformed the strain with a ...

example 3

[0153]To improve metabolism of D-xylose through the D-arabinose degradation pathway, we attempted to overexpress the relevant enzymes: D-ribulokinase (encoded by fucK), D-ribulose-phosphate aldolase (encoded by fucA), and glycolaldehyde reductase (encoded by fucO). These enzymes were overexpressed as part of an operon in conjunction with DTE; the order of the genes in the operon was varied. All of these were grown on D-xylose, analyzed for EG production, and compared against ΔaldA cultured on D-arabinose (FIG. 8). The best strain is ΔaldA ΔxylB / p10_T5T10-dte-fucA-fucO-fucK, which outperforms the D-arabinose results. When a plasmid of a lower copy number was used (p5_T5T10-dte-fucA-fucO-fucK), the results were similar (data not shown).

[0154]To maximize EG production, we next grew our best strain, ΔaldA ΔxylB / p5_T5T10-dte-fucA-fucO-fucK, in a bioreactor. The medium of the initial batch was minimal medium with 30 g / L D-xylose; when nearly all D-xylose was consumed, additional D-xylose ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Electrical resistanceaaaaaaaaaa
Login to View More

Abstract

The invention relates to recombinant cells and their use in the production of ethylene glycol.

Description

RELATED APPLICATIONS[0001]This application claims the benefit under 35 U.S.C. §119(e) of U.S. provisional application No. 61 / 602,322, filed Feb. 23, 2012, which is incorporated by reference herein in its entirety.GOVERNMENT INTEREST[0002]This invention was made with government support under Grant No. DE-AR0000059 awarded by the Department of Energy, Office of ARPA-E. The government has certain rights in this invention.FIELD OF THE INVENTION[0003]The invention relates to the production of ethylene glycol through recombinant gene expression and metabolic engineering.BACKGROUND OF THE INVENTION[0004]Ethylene glycol is an important organic compound commonly used as a precursor to polymers, primarily polyethylene terephthalate (PET) which comprises a significant share of the worlds polymer production. The major end uses of PET are synthetic fibers, commonly referred to as “polyester,” and plastic bottles. For both of these products, the demand is increasing. Another major use of ethylene...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12P7/18C12N15/70
CPCC12P7/18C12N15/70C12Y207/01016C12Y101/01071C12Y102/01052C12Y207/01047C12Y401/02017C12Y501/03C12N9/0006C12N9/0008C12N9/0022C12N9/1096C12N9/1205C12N9/1229C12N9/16C12N9/88C12N9/90C12N15/52C12P2203/00C12Y102/01021C12Y207/01017C12P7/24
Inventor STEPHANOPOULOS, GREGORYPEREIRA, BRIANDE MEY, MARJANDUGAR, DEEPAKAVALOS, JOSE LUIS
Owner MASSACHUSETTS INST OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com