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This lecture covers the quantitative analysis of metabolic pathways, focusing on calculating and estimating missing fluxes, simulating the effects of environmental or genetic changes, identifying critical reactions, and understanding pathway branching points. It delves into metabolic flux balance analysis, steady-state assumptions, central carbon metabolism of E. coli, and the reconstruction of genome-scale metabolic models. The lecture also discusses underdetermined systems, constraint-based modeling, and the knowledge gaps in metabolic networks, emphasizing the importance of genome-scale model reconstruction in understanding complex microbiomes. Additionally, it explores the concept of achieving objectives in metabolic systems and the costs of biosynthesis of cellular components.