Biomanufacturing offers a potentially sustainable alternative to deriving chemicals from fossil fuels. However, traditional biomanufacturing, which uses sugars as feedstocks, competes with food production and yields unfavourable land use changes, so more sustainable options are necessary. Cupriavidus necator is a chemolithoautotrophic bacterium capable of consuming carbon dioxide and hydrogen as sole carbon and energy sources, or formate as the source of both. This autotrophic metabolism potentially makes chemical production using C. necator sustainable and attractive for biomanufacturing. Additionally, C. necator natively fixes carbon in the form of poly-3-hydroxybutyrate, which can be processed to make biodegradable plastic. Recent progress in development of modelling and synthetic biology tools have made C. necator much more usable as a biomanufacturing chassis. However, these tools and applications are often limited by a lack of consideration for the unique physiology and metabolic features of C. necator. As such, further work is required to better understand the intricate mechanisms that allow it to prioritise generalization over specialization. In this review, progress toward physiology-informed engineering of C. necator across several dimensions is critically discussed, and recommendations for moving toward a physiological approach are presented. Arguments for metabolic specialization, more focus on autotrophic fermentation, C. necator-specific synthetic biology tools, and modelling that goes beyond constraints are presented based on analysis of existing literature.

Plastics are indispensable to modern life, but their widespread use has created an environmental crisis due to inefficient waste management. Mixed plastic waste, comprising diverse polymers, presents significant recycling challenges due to the high costs of sorting and processing, leading to ecosystem accumulation and harmful by-product generation. This study addresses this issue by engineering a synthetic bacterial consortium (SBC) designed to degrade mixed plastic monomers. The consortium pairs Escherichia coli Nissle 1917, which uses ethylene glycol (EG), a monomer derived from polyethylene terephthalate (PET), as a carbon source, with Pseudomonas putida KT2440, which metabolizes hexamethylenediamine (HD), a monomer from nylon-6,6, as a nitrogen source. Adaptive evolution of the SBC revealed a novel metabolic interaction where P. putida developed the ability to degrade both EG and HD, while E. coli played a critical role in degrading glycolate, mitigating its by-product toxicity. The evolved cross-feeding pattern enhanced biomass production, metabolic efficiency, and community stability compared to monocultures. The consortium’s performance was validated through flux balance analysis (FBA), high-performance liquid chromatography (HPLC), and growth assays. These findings highlight the potential of cross-feeding SBCs in addressing complex plastic waste, offering a promising avenue for sustainable bioremediation and advancing future polymer degradation strategies.

The growing plastics end-of-life crisis threatens ecosystems and human health globally. Microbial plastic degradation and upcycling have emerged as potential solutions to this complex challenge, but their industrial feasibility and limitations thereon have not been fully characterized. In this perspective paper, we review literature describing both plastic degradation and transformation of plastic monomers into value-added products by microbes. We aim to understand the current feasibility of combining these into a single, closed-loop process. Our analysis shows that microbial plastic degradation is currently the rate-limiting step to “closing the loop”, with reported rates that are orders of magnitude lower than those of pathways to upcycle plastic degradation products. We further find that neither degradation nor upcycling have been demonstrated at rates sufficiently high to justify industrialization at present. As a potential way to address these limitations, we suggest more investigation into mixotrophic approaches, showing that those which leverage the unique properties of plastic degradation products such as ethylene glycol might improve rates sufficiently to motivate industrial process development.

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.

In biology, data are conceptualized using diagrams that capture protein-protein, enzyme-substrate, and regulator-target interactions, among many others. These interaction diagrams are every bit as complicated as wiring diagrams in modern electronic circuits—in many cases, even more so. Yet, in contrast to electronic circuits, living systems must also autonomously reproduce; some part of the “wiring diagram” of life must be devoted to reproducing itself. A greatly simplifying principle in the analysis of these biological wiring diagrams is that of “balanced growth,” in which network flows are balanced according to the requirements of biomass production, leading to the exponential accumulation of cells. In microbial cells, exponential growth greatly simplifies the underlying biochemical networks, because when its mathematical description is combined with kinetic descriptions of underlying enzyme-mediated reactions, macroscale constraints on physiology emerge. As is demonstrated in this tutorial, these constraints are mathematically and conceptually equivalent to Kirchhoff's circuit laws and Ohm's constitutive equation. Consequently, bacterial growth physiology can be approached with the same quantitative rigour as electrical circuit analysis. In this tutorial, this “Ohmics” approach is developed in detail, and its power in simplifying complex physiology is demonstrated through two case studies.