Saccharomyces　cerevisiae　predominantly　ferments　sugar　to　ethanol,　irrespective　of　the　presence　of　oxygen,　which　is　known　as　the　Crabtree-effect.　Traditional　methods　rely　on　static　controls　of　glycolytic　flux　to　make　S.　cerevisiae　Crabtree-negative,　which　are　not　favorable　for　future　biomanufacturing　applications.　Considering　native　metabolic　pathways　typically　harness　dynamic　regulatory　networks,　we　therefore　aim　to　develop　an　alternative　strategy　using　dynamic　regulation　of　the　yeast　central　metabolism　to　generate　Crabtree-negative　S.　cerevisiae.　We　report　that　manipulating　a　single　step　at　sugar　phosphorylation　can　alter　the　mode　of　yeast　metabolism　with　an　attenuated　Crabtree-effect.　By　implementing　catabolite-regulated　sugar　phosphorylation,　the　diauxic　shift　in　budding　yeast　was　effectively　reduced.　The　Crabtree-attenuated　metabolism　in　the　engineered　yeast　was　confirmed　by　multidimensional　characterizations　such　as　cell　morphology,　the　measurements　of　sugar　utilization　rate　and　ethanol　production,　and　transcriptomics.　In　addition,　we　demonstrated　that　the　Crabtree-attenuated　metabolism　could　substantially　improve　the　mitochondrial　synthesis　of　short　branched-chain　fatty　acids　from　amino　acid　catabolism,　and　allow　the　synthesis　and　accumulation　of　retinaldehyde.　Taken　together,　we　present　for　the　first　time　that　manipulation　of　sugar　phosphorylation　can　alter　the　mode　of　yeast　metabolism,　and　the　synthetic　Crabtree-attenuated　yeast　factory　established　here　might　serve　as　a　non-fermentative　biomanufacturing　chassis.