title: Pathway swapping: Toward modular engineering of essential cellular processes.
abstract: 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.
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Niels G A
authors.initials: NG
authors.lastname: Kuijpers
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Daniel
authors.initials: D
authors.lastname: Solis-Escalante
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Marijke A H
authors.initials: MA
authors.lastname: Luttik
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Markus M M
authors.initials: MM
authors.lastname: Bisschops
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Francine J
authors.initials: FJ
authors.lastname: Boonekamp
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: van den Broek
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Jack T
authors.initials: JT
authors.lastname: Pronk
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Daran
authors.affiliation: Department of Biotechnology, Delft University of Technology, 2629 HZ Delft, The Netherlands p.a.s.daran-lapujade@tudelft.nl.
authors.forename: Pascale
authors.identifier: 0000-0002-4097-7831
authors.initials: P
authors.lastname: Daran-Lapujade
mesh_terms: D000431:Ethanol / Q000378:metabolism; D018628:Gene Dosage; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D006019:Glycolysis; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways; D005810:Multigene Family; D010641:Phenotype; D015202:Protein Engineering; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D058615:Synthetic Biology
chemical_list: D000431:Ethanol
keywords: Saccharomyces cerevisiae; glycolysis; modular genomes; pathway swapping