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Mutations in PMR1 s...
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Verhoeven, Maarten D.
(author)
Mutations in PMR1 stimulate xylose isomerase activity and anaerobic growth on xylose of engineered Saccharomyces cerevisiae by influencing manganese homeostasis
- Article/chapterEnglish2017
Publisher, publication year, extent ...
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2017-04-12
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Nature Publishing Group,2017
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Numbers
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LIBRIS-ID:oai:DiVA.org:kth-206684
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https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-206684URI
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https://doi.org/10.1038/srep46155DOI
Supplementary language notes
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Language:English
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Summary in:English
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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QC 20170510
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Combined overexpression of xylulokinase, pentose-phosphate-pathway enzymes and a heterologous xylose isomerase (XI) is required but insufficient for anaerobic growth of Saccharomyces cerevisiae on d-xylose. Single-step Cas9-assisted implementation of these modifications yielded a yeast strain expressing Piromyces XI that showed fast aerobic growth on d-xylose. However, anaerobic growth required a 12-day adaptation period. Xylose-adapted cultures carried mutations in PMR1, encoding a Golgi Ca2+/Mn2+ ATPase. Deleting PMR1 in the parental XI-expressing strain enabled instantaneous anaerobic growth on d-xylose. In pmr1 strains, intracellular Mn2+ concentrations were much higher than in the parental strain. XI activity assays in cell extracts and reconstitution experiments with purified XI apoenzyme showed superior enzyme kinetics with Mn2+ relative to other divalent metal ions. This study indicates engineering of metal homeostasis as a relevant approach for optimization of metabolic pathways involving metal-dependent enzymes. Specifically, it identifies metal interactions of heterologous XIs as an underexplored aspect of engineering xylose metabolism in yeast.
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Lee, Misun
(author)
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Kamoen, Lycka
(author)
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van den Broek, Marcel
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Janssen, Dick B.
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Daran, Jean-Marc G.
(author)
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van Maris, Antonius J. A.KTH,Industriell bioteknologi,Department of Biotechnology, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands;(Swepub:kth)u1n330r1
(author)
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Pronk, Jack T.
(author)
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KTHIndustriell bioteknologi
(creator_code:org_t)
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In:Scientific Reports: Nature Publishing Group72045-2322
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