Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae

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Papapetridis, IoannisDelft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands. (author)

Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae

Article/chapterEnglish2018

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2018-05-15
Oxford University Press,2018
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LIBRIS-ID:oai:DiVA.org:kth-239500
https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-239500URI
https://doi.org/10.1093/femsyr/foy056DOI

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Language:English
Summary in:English

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QC 20181126
Simultaneous fermentation of glucose and xylose can contribute to improved productivity and robustness of yeast-based processes for bioethanol production from lignocellulosic hydrolysates. This study explores a novel laboratory evolution strategy for identifying mutations that contribute to simultaneous utilisation of these sugars in batch cultures of Saccharomyces cerevisiae. To force simultaneous utilisation of xylose and glucose, the genes encoding glucose-6-phosphate isomerase (PGI1) and ribulose-5-phosphate epimerase (RPE1) were deleted in a xylose-isomerase-based xylose-fermenting strain with a modified oxidative pentose-phosphate pathway. Laboratory evolution of this strain in serial batch cultures on glucose-xylose mixtures yielded mutants that rapidly co-consumed the two sugars. Whole-genome sequencing of evolved strains identified mutations in HXK2, RSP5 and GAL83, whose introduction into a non-evolved xylose-fermenting S. cerevisiae strain improved co-consumption of xylose and glucose under aerobic and anaerobic conditions. Combined deletion of HXK2 and introduction of a GAL83(G673T) allele yielded a strain with a 2.5-fold higher xylose and glucose co-consumption ratio than its xylose-fermenting parental strain. These two modifications decreased the time required for full sugar conversion in anaerobic bioreactor batch cultures, grown on 20 g L-1 glucose and 10 g L-1 xylose, by over 24 h. This study demonstrates that laboratory evolution and genome resequencing of microbial strains engineered for forced co-consumption is a powerful approach for studying and improving simultaneous conversion of mixed substrates.

Subject headings and genre

TEKNIK OCH TEKNOLOGIER Industriell bioteknik Annan industriell bioteknik hsv//swe
ENGINEERING AND TECHNOLOGY Industrial Biotechnology Other Industrial Biotechnology hsv//eng
biofuels
yeast
industrial biotechnology
redox engineering
fermentation
pentoses

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Verhoeven, Maarten D.Delft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands. (author)
Wiersma, Sanne J.Delft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands. (author)
Goudriaan, MaaikeDelft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands. (author)
van Maris, Antonius J. A.KTH,Industriell bioteknologi,Delft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands.(Swepub:kth)u1n330r1 (author)
Pronk, Jack T.Delft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands. (author)
Delft Univ Technol, Dept Biotechnol, Maasweg 9, NL-2629 HZ Delft, Netherlands.Industriell bioteknologi (creator_code:org_t)

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In:FEMS yeast research (Print): Oxford University Press18:61567-13561567-1364

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