ATHENA: a dynamic metabolic hub balancing carbon economy and energy supply in microbial synthesis

Diverse chemicals biosynthesis demand distinct precursor acetyl-CoA and energy inputs, yet strategies balancing carbon atom economy and energy generation for varied products remain scarce. Here, we design an autonomous hybrid energy-carbon balancing apparatus (ATHENA) to establish a paradigm for redox-driven, dynamically adaptive regulation of carbon conservation and energy synthesis in Escherichia coli. ATHENA features modularization of acetyl-CoA and NADH synthesis pathways for carbon conservation and energy generation, paired with NADH-responsive positive and negative genetic circuits constructed using the BsRex sensor and antisense RNA. This system enables intracellular redox state-dependent rational metabolic flux allocation to balance carbon atom economy and energy generation. Ultimately, ATHENA significantly improves yields and titers of five products with differential acetyl-CoA and energy demands. In the 5-L bioreactor, acetate yield markedly exceeds its native theoretical yield, while that of mevalonate nearly approaches this theoretical upper limit. This work provides a universal strategy for constructing high carbon-yield microbial chassis cells. Metabolic engineering strategies that balance carbon atom economy and energy generation remain scarce. Here the authors engineer E. coli with an NADH-responsive genetic circuit to regulate carbon-conserving and energy-generating modules, improving yields and titres of diverse products.