The study investigates the role of lysine acetylation in regulating metabolic enzymes in *Salmonella*. It finds that central metabolism enzymes in *Salmonella* are extensively and differentially acetylated in response to various carbon sources, affecting cell growth and metabolic flux. The acetylation status of these enzymes is controlled by a pair of lysine acetyltransferase and deacetylase, Pat and CobB, respectively. Reversible acetylation ensures that cells can quickly sense and adapt to environmental changes by altering reaction rates or directions. The study identifies 235 acetylated peptides in 191 proteins, with about 90% of central metabolic enzymes being acetylated. Acetylation levels vary depending on the carbon source, with greater acetylation observed in glucose-grown cells compared to citrate-grown cells. The physiological impact of acetylation is evident in the growth properties of wild-type, *Δpat*, and *ΔcobB* strains under different carbon sources. In vitro studies demonstrate that acetylation regulates the activities of key enzymes such as GapA, AceA, and AceK, affecting glycolysis and gluconeogenesis. The expression of *pat* and *cobB* genes is also regulated in response to different carbon sources, with higher expression during log phase growth. This work highlights a conserved metabolic regulatory mechanism from bacteria to mammals, where reversible lysine acetylation plays a crucial role in coordinating carbon source utilization and metabolic flux.The study investigates the role of lysine acetylation in regulating metabolic enzymes in *Salmonella*. It finds that central metabolism enzymes in *Salmonella* are extensively and differentially acetylated in response to various carbon sources, affecting cell growth and metabolic flux. The acetylation status of these enzymes is controlled by a pair of lysine acetyltransferase and deacetylase, Pat and CobB, respectively. Reversible acetylation ensures that cells can quickly sense and adapt to environmental changes by altering reaction rates or directions. The study identifies 235 acetylated peptides in 191 proteins, with about 90% of central metabolic enzymes being acetylated. Acetylation levels vary depending on the carbon source, with greater acetylation observed in glucose-grown cells compared to citrate-grown cells. The physiological impact of acetylation is evident in the growth properties of wild-type, *Δpat*, and *ΔcobB* strains under different carbon sources. In vitro studies demonstrate that acetylation regulates the activities of key enzymes such as GapA, AceA, and AceK, affecting glycolysis and gluconeogenesis. The expression of *pat* and *cobB* genes is also regulated in response to different carbon sources, with higher expression during log phase growth. This work highlights a conserved metabolic regulatory mechanism from bacteria to mammals, where reversible lysine acetylation plays a crucial role in coordinating carbon source utilization and metabolic flux.