Allosteric Effectors Outcompete Transcript Levels and Substrate Concentration in Regulating Central Carbon Flux During the Crabtree Effect Transition

ABSTRACT While the metabolic shift in Saccharomyces cerevisiae across the Crabtree effect is well‐documented, the role of allosteric regulation in this transition remains unclear. Here, we investigated allosteric regulation by inducing a growth rate shift from 0.2 to 0.35 h. Our results revealed 35 regulatory interactions across 23 central carbon metabolism reactions, with allosteric effectors explaining 29% of flux changes—surpassing the contributions of transcript abundance and substrate concentration. Additionally, key Crabtree‐responsive reactions' flux changes were co‐regulated by allosteric effectors, transcript abundance, and enzyme turnover numbers. These underscore the significance of allosteric regulation in metabolic adaptation during growth rate transitions in Saccharomyces cerevisiae .