{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Castano-Cerezo S"],"funding":["Agence Nationale de la Recherche (ANR)","Agence Nationale de la Recherche"],"pagination":["5169-5185"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11535393"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["43(21)"],"pubmed_abstract":["Enzymatic parameters are classically determined in vitro, under conditions that are far from those encountered in cells, casting doubt on their physiological relevance. We developed a generic approach combining tools from synthetic and systems biology to measure enzymatic parameters in vivo. In the context of a synthetic carotenoid pathway in Saccharomyces cerevisiae, we focused on a phytoene synthase and three phytoene desaturases, which are difficult to study in vitro. We designed, built, and analyzed a collection of yeast strains mimicking substantial variations in substrate concentration by strategically manipulating the expression of geranyl-geranyl pyrophosphate (GGPP) synthase. We successfully determined in vivo Michaelis-Menten parameters (K<sub>M</sub>, V<sub>max</sub>, and k<sub>"],"journal":["The EMBO journal"],"pubmed_title":["Combining systems and synthetic biology for in vivo enzymology."],"pmcid":["PMC11535393"],"funding_grant_id":["ANR-16-CE11-0022"],"pubmed_authors":["Tholey A","Heux S","Garcia-Alles LF","Bellvert F","Kulyk H","Truan G","Galeote V","Camarasa C","Treitz C","Castano-Cerezo S","Chamas A","Millard P"],"additional_accession":[]},"is_claimable":false,"name":"Combining systems and synthetic biology for in vivo enzymology.","description":"Enzymatic parameters are classically determined in vitro, under conditions that are far from those encountered in cells, casting doubt on their physiological relevance. We developed a generic approach combining tools from synthetic and systems biology to measure enzymatic parameters in vivo. In the context of a synthetic carotenoid pathway in Saccharomyces cerevisiae, we focused on a phytoene synthase and three phytoene desaturases, which are difficult to study in vitro. We designed, built, and analyzed a collection of yeast strains mimicking substantial variations in substrate concentration by strategically manipulating the expression of geranyl-geranyl pyrophosphate (GGPP) synthase. We successfully determined in vivo Michaelis-Menten parameters (K<sub>M</sub>, V<sub>max</sub>, and k<sub>","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Nov","modification":"2026-06-01T20:32:20.416Z","creation":"2025-04-04T11:43:22.654Z"},"accession":"S-EPMC11535393","cross_references":{"pubmed":["39322757"],"doi":["10.1038/s44318-024-00251-w"]}}