The study of metabolic pathways is becoming increasingly important to exploit an integrated, systems-level approach for optimizing a desired cellular property or phenotype. In this context, the integration of genomics data with genetic, metabolic and regulatory models is essential because the systematic design of artificial, biological systems requires the identification of robust building blocks like gene promoters, metabolic pathways or genetic circuits taken from natural organisms, and manipulated to develop ad hoc features. Computational tools allowing precise descriptions of natural pathways might thus allow improving the performance of artificial routes. In this review, we introduce the most recent bioinformatics tools enabling detailed characterizations of metabolic pathways in bacteria from different perspectives.