P. putida displays a versatile metabolism, rapid growth rate with simple nutrient requirements, low energy demand for cell maintenance, high robustness against adverse environmental conditions and toxic compounds, and absence of by-products formation. These properties, which position this microorganism as an excellent chassis for metabolic engineering, are intrinsically tied to the particular architecture of the central carbon metabolism. By applying multi-omic systems biology approaches, we aim at unraveling the underlying rationale behind the wiring of central carbon metabolism in P. putida. The ultimate goal of this research line is twofold: on one hand, we are interested in understanding how this bacterium responds to environmental cues, and, on the other hand, we aim at exploiting and improving its biochemical network for biotechnological applications. To this end, we undertake a deep genetic and metabolic engineering of P. putida KT2440 with the overarching objective of designing a reference chassis for whole-cell biocatalysis.