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Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history

BACKGROUND: The most advanced strains of xylose-fermenting Saccharomyces cerevisiae still utilize xylose far less efficiently than glucose, despite the extensive metabolic and evolutionary engineering applied in their development. Systematic comparison of strains across literature is difficult due t...

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Autores principales: Novy, Vera, Wang, Ruifei, Westman, Johan O., Franzén, Carl Johan, Nidetzky, Bernd
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584037/
https://www.ncbi.nlm.nih.gov/pubmed/28878820
http://dx.doi.org/10.1186/s13068-017-0887-9
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author Novy, Vera
Wang, Ruifei
Westman, Johan O.
Franzén, Carl Johan
Nidetzky, Bernd
author_facet Novy, Vera
Wang, Ruifei
Westman, Johan O.
Franzén, Carl Johan
Nidetzky, Bernd
author_sort Novy, Vera
collection PubMed
description BACKGROUND: The most advanced strains of xylose-fermenting Saccharomyces cerevisiae still utilize xylose far less efficiently than glucose, despite the extensive metabolic and evolutionary engineering applied in their development. Systematic comparison of strains across literature is difficult due to widely varying conditions used for determining key physiological parameters. Here, we evaluate an industrial and a laboratory S. cerevisiae strain, which has the assimilation of xylose via xylitol in common, but differ fundamentally in the history of their adaptive laboratory evolution development, and in the cofactor specificity of the xylose reductase (XR) and xylitol dehydrogenase (XDH). RESULTS: In xylose and mixed glucose–xylose shaken bottle fermentations, with and without addition of inhibitor-rich wheat straw hydrolyzate, the specific xylose uptake rate of KE6-12.A (0.27–1.08 g g(CDW)(−1) h(−1)) was 1.1 to twofold higher than that of IBB10B05 (0.10–0.82 g g(CDW)(−1) h(−1)). KE6-12.A further showed a 1.1 to ninefold higher glycerol yield (0.08–0.15 g g(−1)) than IBB10B05 (0.01–0.09 g g(−1)). However, the ethanol yield (0.30–0.40 g g(−1)), xylitol yield (0.08–0.26 g g(−1)), and maximum specific growth rate (0.04–0.27 h(−1)) were in close range for both strains. The robustness of flocculating variants of KE6-12.A (KE-Flow) and IBB10B05 (B-Flow) was analyzed in high-gravity simultaneous saccharification and co-fermentation. As in shaken bottles, KE-Flow showed faster xylose conversion and higher glycerol formation than B-Flow, but final ethanol titres (61 g L(−1)) and cell viability were again comparable for both strains. CONCLUSIONS: Individual specific traits, elicited by the engineering strategy, can affect global physiological parameters of S. cerevisiae in different and, sometimes, unpredictable ways. The industrial strain background and prolonged evolution history in KE6-12.A improved the specific xylose uptake rate more substantially than the superior XR, XDH, and xylulokinase activities were able to elicit in IBB10B05. Use of an engineered XR/XDH pathway in IBB10B05 resulted in a lower glycerol rather than a lower xylitol yield. However, the strain development programs were remarkably convergent in terms of the achieved overall strain performance. This highlights the importance of comparative strain evaluation to advance the engineering strategies for next-generation S. cerevisiae strain development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-017-0887-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-55840372017-09-06 Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history Novy, Vera Wang, Ruifei Westman, Johan O. Franzén, Carl Johan Nidetzky, Bernd Biotechnol Biofuels Research BACKGROUND: The most advanced strains of xylose-fermenting Saccharomyces cerevisiae still utilize xylose far less efficiently than glucose, despite the extensive metabolic and evolutionary engineering applied in their development. Systematic comparison of strains across literature is difficult due to widely varying conditions used for determining key physiological parameters. Here, we evaluate an industrial and a laboratory S. cerevisiae strain, which has the assimilation of xylose via xylitol in common, but differ fundamentally in the history of their adaptive laboratory evolution development, and in the cofactor specificity of the xylose reductase (XR) and xylitol dehydrogenase (XDH). RESULTS: In xylose and mixed glucose–xylose shaken bottle fermentations, with and without addition of inhibitor-rich wheat straw hydrolyzate, the specific xylose uptake rate of KE6-12.A (0.27–1.08 g g(CDW)(−1) h(−1)) was 1.1 to twofold higher than that of IBB10B05 (0.10–0.82 g g(CDW)(−1) h(−1)). KE6-12.A further showed a 1.1 to ninefold higher glycerol yield (0.08–0.15 g g(−1)) than IBB10B05 (0.01–0.09 g g(−1)). However, the ethanol yield (0.30–0.40 g g(−1)), xylitol yield (0.08–0.26 g g(−1)), and maximum specific growth rate (0.04–0.27 h(−1)) were in close range for both strains. The robustness of flocculating variants of KE6-12.A (KE-Flow) and IBB10B05 (B-Flow) was analyzed in high-gravity simultaneous saccharification and co-fermentation. As in shaken bottles, KE-Flow showed faster xylose conversion and higher glycerol formation than B-Flow, but final ethanol titres (61 g L(−1)) and cell viability were again comparable for both strains. CONCLUSIONS: Individual specific traits, elicited by the engineering strategy, can affect global physiological parameters of S. cerevisiae in different and, sometimes, unpredictable ways. The industrial strain background and prolonged evolution history in KE6-12.A improved the specific xylose uptake rate more substantially than the superior XR, XDH, and xylulokinase activities were able to elicit in IBB10B05. Use of an engineered XR/XDH pathway in IBB10B05 resulted in a lower glycerol rather than a lower xylitol yield. However, the strain development programs were remarkably convergent in terms of the achieved overall strain performance. This highlights the importance of comparative strain evaluation to advance the engineering strategies for next-generation S. cerevisiae strain development. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-017-0887-9) contains supplementary material, which is available to authorized users. BioMed Central 2017-09-04 /pmc/articles/PMC5584037/ /pubmed/28878820 http://dx.doi.org/10.1186/s13068-017-0887-9 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Novy, Vera
Wang, Ruifei
Westman, Johan O.
Franzén, Carl Johan
Nidetzky, Bernd
Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title_full Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title_fullStr Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title_full_unstemmed Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title_short Saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity SSCF: convergence in strain performance despite differences in genetic and evolutionary engineering history
title_sort saccharomyces cerevisiae strain comparison in glucose–xylose fermentations on defined substrates and in high-gravity sscf: convergence in strain performance despite differences in genetic and evolutionary engineering history
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584037/
https://www.ncbi.nlm.nih.gov/pubmed/28878820
http://dx.doi.org/10.1186/s13068-017-0887-9
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