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Biochemical pathway and LC-based in vivo screen for thioesterases with improved substrate specificity. The pathway in E. coli for the biosynthesis of SCFAs contains enzymes Pct (Megaphaera elsdenii), BktB (Cupriavidus necator), PhaB (Cupriavidus necator), PhaJ4 (Cupriavidus necator), and Ter (Treponema denticola) with varied thioesterases. The dashed bond indicates the additional carbon incorporated into the fatty acids with feeding of isobutyrate in place of propionate.
