Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase
Over the past two decades, significant progress has been made in the engineering of xylose-consuming Saccharomyces cerevisiae strains for production of lignocellulosic biofuels. However, the ethanol productivities achieved on xylose are still significantly lower than those observed on glucose for re...
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Wiley Blackwell
2017
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Online Access: | http://hdl.handle.net/1721.1/109764 https://orcid.org/0000-0001-6909-4568 |
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author | Wasylenko, Thomas M. Stephanopoulos, Gregory Wasylenko, Thomas Michael |
author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Wasylenko, Thomas M. Stephanopoulos, Gregory Wasylenko, Thomas Michael |
author_sort | Wasylenko, Thomas M. |
collection | MIT |
description | Over the past two decades, significant progress has been made in the engineering of xylose-consuming Saccharomyces cerevisiae strains for production of lignocellulosic biofuels. However, the ethanol productivities achieved on xylose are still significantly lower than those observed on glucose for reasons that are not well understood. We have undertaken an analysis of central carbon metabolite pool sizes and metabolic fluxes on glucose and on xylose under aerobic and anaerobic conditions in a strain capable of rapid xylose assimilation via xylose isomerase in order to investigate factors that may limit the rate of xylose fermentation. We find that during xylose utilization the flux through the non-oxidative Pentose Phosphate Pathway (PPP) is high but the flux through the oxidative PPP is low, highlighting an advantage of the strain employed in this study. Furthermore, xylose fails to elicit the full carbon catabolite repression response that is characteristic of glucose fermentation in S. cerevisiae. We present indirect evidence that the incomplete activation of the fermentation program on xylose results in a bottleneck in lower glycolysis, leading to inefficient re-oxidation of NADH produced in glycolysis. |
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id | mit-1721.1/109764 |
institution | Massachusetts Institute of Technology |
language | en_US |
last_indexed | 2024-09-23T11:44:38Z |
publishDate | 2017 |
publisher | Wiley Blackwell |
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spelling | mit-1721.1/1097642022-09-27T21:38:03Z Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase Metabolomic and 13C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase Wasylenko, Thomas M. Stephanopoulos, Gregory Wasylenko, Thomas Michael Massachusetts Institute of Technology. Department of Chemical Engineering Wasylenko, Thomas Michael Stephanopoulos, Gregory Over the past two decades, significant progress has been made in the engineering of xylose-consuming Saccharomyces cerevisiae strains for production of lignocellulosic biofuels. However, the ethanol productivities achieved on xylose are still significantly lower than those observed on glucose for reasons that are not well understood. We have undertaken an analysis of central carbon metabolite pool sizes and metabolic fluxes on glucose and on xylose under aerobic and anaerobic conditions in a strain capable of rapid xylose assimilation via xylose isomerase in order to investigate factors that may limit the rate of xylose fermentation. We find that during xylose utilization the flux through the non-oxidative Pentose Phosphate Pathway (PPP) is high but the flux through the oxidative PPP is low, highlighting an advantage of the strain employed in this study. Furthermore, xylose fails to elicit the full carbon catabolite repression response that is characteristic of glucose fermentation in S. cerevisiae. We present indirect evidence that the incomplete activation of the fermentation program on xylose results in a bottleneck in lower glycolysis, leading to inefficient re-oxidation of NADH produced in glycolysis. Shell Oil Company National Institute of General Medical Sciences (U.S.) Biotechnology Training Program 2017-06-09T14:44:43Z 2017-06-09T14:44:43Z 2014-11 2014-08 Article http://purl.org/eprint/type/JournalArticle 0006-3592 http://hdl.handle.net/1721.1/109764 Wasylenko, Thomas M., and Gregory Stephanopoulos. “Metabolomic and 13 C-Metabolic Flux Analysis of a Xylose-Consuming Saccharomyces Cerevisiae Strain Expressing Xylose Isomerase: Xylose Metabolic Flux Analysis.” Biotechnology and Bioengineering 112.3 (2015): 470–483. https://orcid.org/0000-0001-6909-4568 en_US http://dx.doi.org/10.1002/bit.25447 Biotechnology and Bioengineering Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley Blackwell PMC |
spellingShingle | Wasylenko, Thomas M. Stephanopoulos, Gregory Wasylenko, Thomas Michael Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title | Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title_full | Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title_fullStr | Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title_full_unstemmed | Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title_short | Metabolomic and [superscript 13]C-metabolic flux analysis of a xylose-consuming Saccharomyces cerevisiae strain expressing xylose isomerase |
title_sort | metabolomic and superscript 13 c metabolic flux analysis of a xylose consuming saccharomyces cerevisiae strain expressing xylose isomerase |
url | http://hdl.handle.net/1721.1/109764 https://orcid.org/0000-0001-6909-4568 |
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