NDLI: Development of a D-xylose fermenting and inhibitor tolerant industrial Saccharomyces cerevisiae strain with high performance in lignocellulose hydrolysates using metabolic and evolutionary engineering

Content Provider	Springer Nature : BioMed Central
Author	Demeke, Mekonnen M Dietz, Heiko Li, Yingying Foulquié-Moreno, María R Mutturi, Sarma Deprez, Sylvie Den Abt, Tom Bonini, Beatriz M Liden, Gunnar Dumortier, Françoise Verplaetse, Alex Boles, Eckhard Thevelein, Johan M
Abstract	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.
Related Links	https://biotechnologyforbiofuels.biomedcentral.com/counter/pdf/10.1186/1754-6834-6-89.pdf
Ending Page	24
Page Count	24
Starting Page	1
File Format	HTM / HTML
ISSN	27313654
DOI	10.1186/1754-6834-6-89
Journal	Biotechnology for Biofuels and Bioproducts
Issue Number	1
Volume Number	6
Language	English
Publisher	BioMed Central
Publisher Date	2013-06-21
Access Restriction	Open
Subject Keyword	Biotechnology Plant Breeding Environmental Engineering Renewable and Green Energy Microbiology Saccharomyces cerevisiae Bioethanol Lignocellulose D-xylose fermentation D-xylose isomerase Inhibitor tolerance Evolutionary engineering Plant Breeding/Biotechnology Environmental Engineering/Biotechnology
Content Type	Text
Resource Type	Article
Subject	Energy Management, Monitoring, Policy and Law Renewable Energy, Sustainability and the Environment Applied Microbiology and Biotechnology Biotechnology
Journal Impact Factor	6.1/2023
5-Year Journal Impact Factor	6/2023

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