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Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism
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2017 (English)In: FEMS yeast research (Print), ISSN 1567-1356, E-ISSN 1567-1364, Vol. 17, no 1, fox006Article in journal (Refereed) Published
Abstract [en]

Many relevant options to improve efficacy and kinetics of sucrose metabolism in Saccharomyces cerevisiae and, thereby, the economics of sucrose-based processes remain to be investigated. An essential first step is to identify all native sucrose-hydrolysing enzymes and sucrose transporters in this yeast, including those that can be activated by suppressor mutations in sucrose-negative strains. A strain in which all known sucrose-transporter genes (MAL11, MAL21, MAL31, MPH2, MPH3) were deleted did not grow on sucrose after 2 months of incubation. In contrast, a strain with deletions in genes encoding sucrose-hydrolysing enzymes (SUC2, MAL12, MAL22, MAL32) still grew on sucrose. Its specific growth rate increased from 0.08 to 0.25 h(-1) after sequential batch cultivation. This increase was accompanied by a 3-fold increase of in vitro sucrose-hydrolysis and isomaltase activities, as well as by a 3- to 5-fold upregulation of the isomaltase-encoding genes IMA1 and IMA5. One-step Cas9-mediated deletion of all isomaltase-encoding genes (IMA1-5) completely abolished sucrose hydrolysis. Even after 2 months of incubation, the resulting strain did not grow on sucrose. This sucrose-negative strain can be used as a platform to test metabolic engineering strategies and for fundamental studies into sucrose hydrolysis or transport.

Place, publisher, year, edition, pages
Oxford University Press, 2017. Vol. 17, no 1, fox006
Keyword [en]
disaccharide, isomaltase, laboratory evolution, reverse engineering, multiple gene deletion, real-time PCR
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:kth:diva-211631DOI: 10.1093/femsyr/fox006ISI: 000405634600015OAI: oai:DiVA.org:kth-211631DiVA: diva2:1130434
Note

QC 20170809

Available from: 2017-08-09 Created: 2017-08-09 Last updated: 2017-12-07Bibliographically approved

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van Maris, Antonius J. A.

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