TY - JOUR
T1 - Pathway swapping
T2 - Toward modular engineering of essential cellular processes
AU - Kuijpers, Niels G.A.
AU - Solis-Escalante, Daniel
AU - Luttik, Marijke A.H.
AU - Bisschops, Markus M.M.
AU - Boonekamp, Francine J.
AU - Van Den Broek, Marcel
AU - Pronk, Jack T.
AU - Daran, Jean Marc
AU - Daran-Lapujade, Pascale
PY - 2016/12/27
Y1 - 2016/12/27
N2 - Recent developments in synthetic biology enable one-step implementation of entire metabolic pathways in industrial microorganisms. A similarly radical remodelling of central metabolism could greatly accelerate fundamental and applied research, but is impeded by the mosaic organization of microbial genomes. To eliminate this limitation, we propose and explore the concept of "pathway swapping," using yeast glycolysis as the experimental model. Construction of a "single-locus glycolysis" Saccharomyces cerevisiae platform enabled quick and easy replacement of this yeast's entire complement of 26 glycolytic isoenzymes by any alternative, functional glycolytic pathway configuration. The potential of this approach was demonstrated by the construction and characterization of S. cerevisiae strains whose growth depended on two nonnative glycolytic pathways: a complete glycolysis from the related yeast Saccharomyces kudriavzevii and a mosaic glycolysis consisting of yeast and human enzymes. This work demonstrates the feasibility and potential of modular, combinatorial approaches to engineering and analysis of core cellular processes.
AB - Recent developments in synthetic biology enable one-step implementation of entire metabolic pathways in industrial microorganisms. A similarly radical remodelling of central metabolism could greatly accelerate fundamental and applied research, but is impeded by the mosaic organization of microbial genomes. To eliminate this limitation, we propose and explore the concept of "pathway swapping," using yeast glycolysis as the experimental model. Construction of a "single-locus glycolysis" Saccharomyces cerevisiae platform enabled quick and easy replacement of this yeast's entire complement of 26 glycolytic isoenzymes by any alternative, functional glycolytic pathway configuration. The potential of this approach was demonstrated by the construction and characterization of S. cerevisiae strains whose growth depended on two nonnative glycolytic pathways: a complete glycolysis from the related yeast Saccharomyces kudriavzevii and a mosaic glycolysis consisting of yeast and human enzymes. This work demonstrates the feasibility and potential of modular, combinatorial approaches to engineering and analysis of core cellular processes.
KW - Glycolysis
KW - Modular genomes
KW - Pathway swapping
KW - Saccharomyces cerevisiae
UR - http://www.scopus.com/inward/record.url?scp=85007416022&partnerID=8YFLogxK
U2 - 10.1073/pnas.1606701113
DO - 10.1073/pnas.1606701113
M3 - Article
SN - 0027-8424
VL - 113
SP - 15060
EP - 15065
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 52
ER -