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Title: Engineering microorganisms to increase ethanol production by metabolic redirection

Abstract

The present invention provides for the manipulation of carbon flux in a recombinant host cell to increase the formation of desirable products. The invention relates to cellulose-digesting organisms that have been genetically modified to allow the production of ethanol at a high yield by redirecting carbon flux at key steps of central metabolism.

Inventors:
; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Enchi Corporation, Wellesley Hills, MA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1406573
Patent Number(s):
9,803,221
Application Number:
14/348,231
Assignee:
Enchi Corporation GFO
DOE Contract Number:
FC36-07GO17057; POS2-06ER64304
Resource Type:
Patent
Resource Relation:
Patent File Date: 2012 Sep 28
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 09 BIOMASS FUELS

Citation Formats

Deng, Yu, Olson, Daniel G., van Dijken, Johannes Pieter, Shaw, IV, Arthur J., Argyros, Aaron, Barrett, Trisha, Caiazza, Nicky, Herring, Christopher D., Rogers, Stephen R., and Agbogbo, Frank. Engineering microorganisms to increase ethanol production by metabolic redirection. United States: N. p., 2017. Web.
Deng, Yu, Olson, Daniel G., van Dijken, Johannes Pieter, Shaw, IV, Arthur J., Argyros, Aaron, Barrett, Trisha, Caiazza, Nicky, Herring, Christopher D., Rogers, Stephen R., & Agbogbo, Frank. Engineering microorganisms to increase ethanol production by metabolic redirection. United States.
Deng, Yu, Olson, Daniel G., van Dijken, Johannes Pieter, Shaw, IV, Arthur J., Argyros, Aaron, Barrett, Trisha, Caiazza, Nicky, Herring, Christopher D., Rogers, Stephen R., and Agbogbo, Frank. 2017. "Engineering microorganisms to increase ethanol production by metabolic redirection". United States. doi:. https://www.osti.gov/servlets/purl/1406573.
@article{osti_1406573,
title = {Engineering microorganisms to increase ethanol production by metabolic redirection},
author = {Deng, Yu and Olson, Daniel G. and van Dijken, Johannes Pieter and Shaw, IV, Arthur J. and Argyros, Aaron and Barrett, Trisha and Caiazza, Nicky and Herring, Christopher D. and Rogers, Stephen R. and Agbogbo, Frank},
abstractNote = {The present invention provides for the manipulation of carbon flux in a recombinant host cell to increase the formation of desirable products. The invention relates to cellulose-digesting organisms that have been genetically modified to allow the production of ethanol at a high yield by redirecting carbon flux at key steps of central metabolism.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = 2017,
month =
}

Patent:

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  • The ability to generate microorganisms that can produce biofuels similar to petroleum-based transportation fuels would allow the use of existing engines and infrastructure and would save an enormous amount of capital required for replacing the current infrastructure to accommodate biofuels that have properties significantly different from petroleum-based fuels. Several groups have demonstrated the feasibility of manipulating microbes to produce molecules similar to petroleum-derived products, albeit at relatively low productivity (e.g. maximum butanol production is around 20 g/L). For cost-effective production of biofuels, the fuel-producing hosts and pathways must be engineered and optimized. Advances in metabolic engineering and synthetic biology willmore » provide new tools for metabolic engineers to better understand how to rewire the cell in order to create the desired phenotypes for the production of economically viable biofuels.« less
  • The efficient diversion of pyruvate from normal fermentative pathways to ethanol production in Klebsiella oxytoca M5A1 requires the expression of Zymomanas mobilis genes encoding both pyruvate decarboxylase and alcohol dehydrogenase. Final ethanol concentrations obtained with the best recombinant, strain M5A1 (pLOI555), were in excess of 40 g/liter with an efficiency of 0.48 g of ethanol (xylose) and 0.50 g of ethanol (glucose) per g of sugar, as compared with a theoretical maximum of 0.51 of ethanol per g of sugar. The maximal volumetric productivity per hour for both sugars was 2.0 g/liter. This volumetric productivity with xylose is almost twicemore » that previously obtained with ethanologenic Escherichia coli. Succinate was also produced as a minor product during fermentation.« less
  • Ethanol is an important industrial chemical and a national effort directed towards its development as an alternative transportation fuel will be successful if it can be produced from renewable feedstocks at economical cost. These feedstocks, such as agricultural and forestry residues, typically contain cellulose, hemicellulose and lignin. Unfortunately, conversion of pentoses sugars in the hemicellulose fraction, and particularly xylose, has been difficult due to the lack of a suitable biocatalyst. Zymomonas mobilis has demonstrated several advantages for ethanol production from glucose but is unable to ferment xylose due to the lack of xylose assimilation and pentose metabolism pathways. To broadenmore » its range of fermentable sugars, we have metabolically engineered Z. mobilis to grow on and efficiently ferment xylose to ethanol by introducing the required metabolic pathways. The engineered strain efficiently ferments both glucose and xylose, which is essential for economical conversion of lignocellulosic biomass to ethanol.« less
  • Technologies are available which will allow the conversion of lignocellulose into fuel ethanol using genetically engineered bacteria. Assembling these into a cost-effective process remains a challenge. The authors` work has focused primarily on the genetic engineering of enteric bacteria using a portable ethanol production pathway. Genes encoding Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase have been integrated into the chromosome of Escherichia coli B to produce strain KO11 for the fermentation of hemicellulose-derived syrups. This organism can efficiently ferment all hexose and pentose sugars present in the polymers of hemicellulose. Klebsiella oxytoca M5A1 has been genetically engineered in a similarmore » manner to produce strain P2 for ethanol production from cellulose. This organism has the native ability to ferment cellobiose and cellotriose, eliminating the need for one class of cellulase enzymes. The optimal pH for cellulose fermentation with this organism is near that of fungal cellulases. The general approach for the genetic engineering of new biocatalysts has been most successful with enteric bacteria thus far. However, this approach may also prove useful with gram-positive bacteria which have other important traits for lignocellulose conversion. Many opportunities remain for further improvements in the biomass to ethanol processes.« less