Core metabolic model for Cupriavidus necator.

Modern chemical manufacturing, on which human quality of life depends, is unsustainable; alternative production routes must be developed. Electrochemical and biological processes offer promise for upgrading waste streams, including recalcitrant carbon dioxide and plastic-derived wastes. However, the inherent heterogeneity and high energy requirements of upcycling the chemical endpoints of the “take-make-waste" economy remain challenging. Cupriavidus necator is an emergent catalyst for complex feedstock valorization because of its extreme metabolic flexibility, which allows it to utilize a wide array of substrates, and its ability to use carbon dioxide via the Calvin-Benson-Bassham cycle. C. necator natively oxidizes hydrogen to power carbon utilization, but its flexibility offers an as-yet unexplored opportunity to couple waste stream oxidation with carbon dioxide utilization instead, potentially enabling carbon conservative waste upcycling. Here, we uncover the constraints on carbon conservative chemical transformation using C. necator as a model. We systematically examine the carbon yield and thermodynamic feasibility of mixotrophic scenarios combining waste-derived carbon sources with hydrogen oxidation to power carbon reassimilation. Then, we evaluate carbon-carbon mixotrophic scenarios, with one carbon source providing electrons in place of hydrogen oxidation. We show that both hydrogen and ethylene glycol are feasible electron sources to drive carbon-neutral or carbon-negative mixotrophic upgrading of waste streams such as acetate or butyrate. In contrast, we find that carbon conservation is likely infeasible for most other waste-derived carbon sources. This work provides a roadmap to establishing novel C. necator strains capable of carbon efficient waste upcycling.