Mekonnen M. Demeke, Heiko Dietz, Yingying Li, María R. Foulquié-Moreno, Sarma Mutturi, Sylvie Deprez, Tom Den Abt, Beatriz M. Bonini, Gunnar Liden, Françoise Dumortier, Alex Verplaetse, Eckhard Boles, Johan M. Thevelein
- Background: The production of bioethanol from lignocellulose hydrolysates requires a robust, D-xylose-fermenting and inhibitor-tolerant microorganism as catalyst. The purpose of the present work was to develop such a strain from a prime industrial yeast strain, Ethanol Red, used for bioethanol production.
Results: An expression cassette containing 13 genes including Clostridium phytofermentans XylA, encoding D-xylose isomerase (XI), and enzymes of the pentose phosphate pathway was inserted in two copies in the genome of Ethanol Red. Subsequent EMS mutagenesis, genome shuffling and selection in D-xylose-enriched lignocellulose hydrolysate, followed by multiple rounds of evolutionary engineering in complex medium with D-xylose, gradually established efficient D-xylose fermentation. The best-performing strain, GS1.11-26, showed a maximum specific D-xylose consumption rate of 1.1 g/g DW/h in synthetic medium, with complete attenuation of 35 g/L D-xylose in about 17 h. In separate hydrolysis and fermentation of lignocellulose hydrolysates of Arundo donax (giant reed), spruce and a wheat straw/hay mixture, the maximum specific D-xylose consumption rate was 0.36, 0.23 and 1.1 g/g DW inoculum/h, and the final ethanol titer was 4.2, 3.9 and 5.8% (v/v), respectively. In simultaneous saccharification and fermentation of Arundo hydrolysate, GS1.11-26 produced 32% more ethanol than the parent strain Ethanol Red, due to efficient D-xylose utilization. The high D-xylose fermentation capacity was stable after extended growth in glucose. Cell extracts of strain GS1.11-26 displayed 17-fold higher XI activity compared to the parent strain, but overexpression of XI alone was not enough to establish D-xylose fermentation. The high D-xylose consumption rate was due to synergistic interaction between the high XI activity and one or more mutations in the genome. The GS1.11-26 had a partial respiratory defect causing a reduced aerobic growth rate.
Conclusions: An industrial yeast strain for bioethanol production with lignocellulose hydrolysates has been developed in the genetic background of a strain widely used for commercial bioethanol production. The strain uses glucose and D-xylose with high consumption rates and partial cofermentation in various lignocellulose hydrolysates with very high ethanol yield. The GS1.11-26 strain shows highly promising potential for further development of an all-round robust yeast strain for efficient fermentation of various lignocellulose hydrolysates.
MetadatenAuthor: | Mekonnen M. Demeke, Heiko Dietz, Yingying Li, María R. Foulquié-Moreno, Sarma Mutturi, Sylvie Deprez, Tom Den Abt, Beatriz M. Bonini, Gunnar Liden, Françoise Dumortier, Alex Verplaetse, Eckhard BolesORCiD, Johan M. Thevelein |
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URN: | urn:nbn:de:hebis:30:3-315114 |
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DOI: | https://doi.org/10.1186/1754-6834-6-89 |
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ISSN: | 1754-6834 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/23800147 |
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Parent Title (English): | Biotechnology for biofuels |
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Publisher: | BioMed Central |
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Place of publication: | London |
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Document Type: | Article |
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Language: | English |
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Date of Publication (online): | 2013/06/21 |
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Date of first Publication: | 2013/06/21 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2013/08/23 |
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Tag: | Bioethanol; D-xylose fermentation; D-xylose isomerase; Evolutionary engineering; Inhibitor tolerance; Lignocellulose; Saccharomyces cerevisiae |
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Volume: | 6 |
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Issue: | 89 |
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Page Number: | 24 |
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Note: | © 2013 Demeke et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
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HeBIS-PPN: | 352730811 |
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Institutes: | Biowissenschaften / Biowissenschaften |
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Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
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Sammlungen: | Universitätspublikationen |
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| Sammlung Biologie / Sondersammelgebiets-Volltexte |
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Licence (German): | Creative Commons - Namensnennung 2.0 |
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