Combining systems and synthetic biology for in vivo enzymology.

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_M, V_max, and k

SUBMITTER: Castano-Cerezo S 

PROVIDER: S-EPMC11535393 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature