Print Email Facebook Twitter A versatile in situ cofactor enhancing system for meeting cellular demands for engineered metabolic pathways Title A versatile in situ cofactor enhancing system for meeting cellular demands for engineered metabolic pathways Author Jaroensuk, Juthamas (Vidyasirimedhi Institute of Science and Technology) Sutthaphirom, Chalermroj (Vidyasirimedhi Institute of Science and Technology) Phonbuppha, Jittima (Vidyasirimedhi Institute of Science and Technology) Chinantuya, Wachirawit (Vidyasirimedhi Institute of Science and Technology; Mahidol University) Kesornpun, Chatchai (Vidyasirimedhi Institute of Science and Technology) Akeratchatapan, Nattanon (Vidyasirimedhi Institute of Science and Technology) Kittipanukul, Narongyot (Vidyasirimedhi Institute of Science and Technology) Phatinuwat, Kamonwan (Chulabhorn Graduate Institute, Bangkok) Hollmann, F. (TU Delft BT/Biocatalysis) Date 2024 Abstract Cofactor imbalance obstructs the productivities of metabolically engineered cells. Herein, we employed a minimally perturbing system, xylose reductase and lactose (XR/lactose), to increase the levels of a pool of sugar phosphates which are connected to the biosynthesis of NAD(P)H, FAD, FMN, and ATP in Escherichia coli. The XR/lactose system could increase the amounts of the precursors of these cofactors and was tested with three different metabolically engineered cell systems (fatty alcohol biosynthesis, bioluminescence light generation, and alkane biosynthesis) with different cofactor demands. Productivities of these cells were increased 2-4-fold by the XR/lactose system. Untargeted metabolomic analysis revealed different metabolite patterns among these cells, demonstrating that only metabolites involved in relevant cofactor biosynthesis were altered. The results were also confirmed by transcriptomic analysis. Another sugar reducing system (glucose dehydrogenase) could also be used to increase fatty alcohol production but resulted in less yield enhancement than XR. This work demonstrates that the approach of increasing cellular sugar phosphates can be a generic tool to increase in vivo cofactor generation upon cellular demand for synthetic biology. Subject alkanebioluminescencefatty alcoholmetabolic engineeringsugar phosphatesynthetic biologyxylose reductase To reference this document use: http://resolver.tudelft.nl/uuid:01775ed6-97c1-4264-9ec8-a686e195a771 DOI https://doi.org/10.1016/j.jbc.2023.105598 ISSN 0021-9258 Source Journal of Biological Chemistry, 300 (2) Part of collection Institutional Repository Document type journal article Rights © 2024 Juthamas Jaroensuk, Chalermroj Sutthaphirom, Jittima Phonbuppha, Wachirawit Chinantuya, Chatchai Kesornpun, Nattanon Akeratchatapan, Narongyot Kittipanukul, Kamonwan Phatinuwat, F. Hollmann, More Authors Files PDF PIIS0021925823026273.pdf 4.37 MB Close viewer /islandora/object/uuid:01775ed6-97c1-4264-9ec8-a686e195a771/datastream/OBJ/view