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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae: a platform strain for engineering sucrose metabolism

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Author(s):
Marques, Wesley Leoricy ; Mans, Robert ; Marella, Eko Roy ; Cordeiro, Rosa Lorizolla ; van den Broek, Marcel ; Daran, Jean-Marc G. ; Pronk, Jack T. ; Gombert, Andreas K. ; van Maris, Antonius J. A.
Total Authors: 9
Document type: Journal article
Source: FEMS Yeast Research; v. 17, n. 1 JAN 2017.
Web of Science Citations: 8
Abstract

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. (AU)

FAPESP's process: 12/05548-1 - Metabolic engineering of Saccharomyces cerevisiae aimed at improving the energetic yield of sucrose metabolism
Grantee:Wesley Leoricy Marques
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 14/07962-5 - Role of isomaltases during sucrose metabolism in Saccharomyces cerevisiae
Grantee:Rosa Lorizolla Cordeiro
Support Opportunities: Scholarships in Brazil - Scientific Initiation