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

Autor(es)

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.

• Tipo de documento: Artigo Científico
• Fonte: FEMS Yeast Research; v. 17, n. 1 JAN 2017.
• Citações Web of Science: 8

Resumo

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)

• Processo FAPESP: 12/05548-1 - Engenharia metabólica de Saccharomyces cerevisiae para o aumento do rendimento energético do metabolismo da sacarose
• Beneficiário: Wesley Leoricy Marques
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 14/07962-5 - Investigação do papel das isomaltases no metabolismo de sacarose em Saccharomyces cerevisiae
• Beneficiário: Rosa Lorizolla Cordeiro
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica