Print Email Facebook Twitter Full humanization of the glycolytic pathway in Saccharomyces cerevisiae Title Full humanization of the glycolytic pathway in Saccharomyces cerevisiae Author Boonekamp, F.J. (TU Delft BT/Industriele Microbiologie) Knibbe, E. (TU Delft BT/Industriele Microbiologie) Vieira Lara, M.A. (University Medical Center Groningen) Wijsman, M. (TU Delft BT/Industriele Microbiologie) Luttik, M.A.H. (TU Delft BT/Industriele Microbiologie) van Eunen, Karen (University Medical Center Groningen) den Ridder, M.J. (TU Delft BT/Industriele Microbiologie) Pabst, Martin (TU Delft BT/Environmental Biotechnology) Daran, J.G. (TU Delft BT/Industriele Microbiologie) Daran-Lapujade, P.A.S. (TU Delft BT/Industriele Microbiologie) Date 2022 Abstract Although transplantation of single genes in yeast plays a key role in elucidating gene functionality in metazoans, technical challenges hamper humanization of full pathways and processes. Empowered by advances in synthetic biology, this study demonstrates the feasibility and implementation of full humanization of glycolysis in yeast. Single gene and full pathway transplantation revealed the remarkable conservation of glycolytic and moonlighting functions and, combined with evolutionary strategies, brought to light context-dependent responses. Human hexokinase 1 and 2, but not 4, required mutations in their catalytic or allosteric sites for functionality in yeast, whereas hexokinase 3 was unable to complement its yeast ortholog. Comparison with human tissues cultures showed preservation of turnover numbers of human glycolytic enzymes in yeast and human cell cultures. This demonstration of transplantation of an entire essential pathway paves the way for establishment of species-, tissue-, and disease-specific metazoan models. Subject adaptive laboratory evolutionCP: Metabolismglycolysishexokinasehumanized yeast modelmoonlighting functionpathway transplantationSaccharomyces cerevisiaesynthetic biology To reference this document use: http://resolver.tudelft.nl/uuid:77cfe001-7fc5-4d09-bf10-2604917b40c3 DOI https://doi.org/10.1016/j.celrep.2022.111010 ISSN 2211-1247 Source Cell Reports, 39 (13), 111010 Part of collection Institutional Repository Document type journal article Rights © 2022 F.J. Boonekamp, E. Knibbe, M.A. Vieira Lara, M. Wijsman, M.A.H. Luttik, Karen van Eunen, M.J. den Ridder, Martin Pabst, J.G. Daran, P.A.S. Daran-Lapujade Files PDF 1_s2.0_S2211124722007999_main.pdf 4.92 MB Close viewer /islandora/object/uuid:77cfe001-7fc5-4d09-bf10-2604917b40c3/datastream/OBJ/view