%0 Generic
%A Knibbe, Ewout
%A Boonekamp, Francine J.
%A Stuij, Rachel
%A Pelsma, Koen A.J.
%A Jansen, Liset
%A Flores, Carmen-Lisset
%A Daran-Lapujade, Pascale
%D 2022
%T Supplementary information to: Simpler is not always better: transplanting the Yarrowia lipolytica glycolytic pathway into Saccharomyces cerevisiae reveals essential synergetic regulatory mechanisms
%U https://data.4tu.nl/articles/dataset/Supplementary_information_to_Simpler_is_not_always_better_transplanting_the_Yarrowia_lipolytica_glycolytic_pathway_into_Saccharomyces_cerevisiae_reveals_essential_synergetic_regulatory_mechanisms/19228347/1
%R 10.4121/19228347.v1
%K Saccharomyces cerevisiae
%K Pathway transplantation
%K Glycolysis
%K Metabolic regulation
%K Bistability
%K hexokinase
%K Yarrowia lipolytica
%X This document contains supplementary Tables and Figures pertaining to the research article entitled: Simpler is not always better: transplanting the <i>Yarrowia lipolytica</i> glycolytic pathway into<i> Saccharomyces cerevisiae</i> reveals essential synergetic regulatory mechanisms.<br><h2>Abstract </h2>

The Embden-Meyerof-Parnas pathway of glycolysis
is a widely distributed and intensively investigated metabolic route. While allosteric
regulation is thought to be essential for glycolytic flux dynamics in many
organisms including yeast, to date single enzyme complementation studies with
non-allosteric glycolytic enzymes have failed to experimentally demonstrate
this essentiality and quantify the overall contribution of allosteric
regulation in tuning the glycolytic flux. This study brings new insight in the
synergetic metabolic role of allosteric regulation by implementing pathway
swapping, a strategy enabling to remodel, in two simple genetic interventions,
the entire glycolytic pathway of <i>Saccharomyces
cerevisiae.</i> <i>S. cerevisiae</i> equipped
with the full set of non-allosteric glycolytic enzymes from the oleaginous yeast
<i>Y. lipolytica</i> lost the ability to
grow on media containing 2% glucose and displayed dynamic responses suggesting metabolic
imbalance between upper and lower glycolysis. Single and combined gene
complementation demonstrated that this phenotype was caused by the simultaneous
deregulation of the three key kinases: hexokinase, phosphofructokinase and
pyruvate kinase. ‘Deregulated glycolysis’ <i>S.
cerevisiae</i> strains could naturally restore glycolytic stability and growth
on glucose by evolving mutations in the <i>Y.
lipolytica</i> glucokinase, causing a strong decrease in glucokinase activity
and glycolytic flux. This solution could be recapitulated in non-evolved
deregulated glycolysis <i>S. cerevisiae</i>
strains by experimentally tuning glucose import. Supported by kinetic
modelling, the present work demonstrates the major synergetic role played by
allosteric regulations in preventing metabolic imbalance in glycolysis and
highlights the power of synthetic biology in addressing long-standing questions
in systems biology.<br>
%I 4TU.ResearchData