Third-generation (3G) biorefineries harnessing industrial off-gases have received significant attention in the transition towards a sustainable circular economy. However, uncertainties surrounding their techno-economic feasibility are hampering widespread commercialization to date. This study investigates the production of single-cell protein (SCP), a sustainable alternative food and feed protein, from steel mill off-gas through an efficient coupled fermentation approach utilizing acetate as an intermediate. A comprehensive model that comprises both the gas-to-acetate and the acetate-to-SCP fermentation processes, as well as gas pretreatment and downstream processing (DSP) operations, was developed and used to perform a techno-economic analysis (TEA). Sensitivity analyses demonstrated that significant cost reductions can be achieved by the process intensification of the gas-to-acetate fermentation. As such, an increase in the acetate concentration to 45 g/L and productivity to 4 g/L/h could lead to a potential cost reduction from 4.15 to 2.78 USD/kg. In addition, the influence of the production scale and other economic considerations towards the commercialization of off-gas-based SCPs are discussed. Conclusively, this research sheds light on the practical viability of a coupled fermentation process for SCP production by identifying key cost-influencing factors and providing targets for further optimization of the acetate platform, fostering sustainable and economically feasible bio-based innovations.

Over the past decade, formic acid and acetic acid have gained increasing attention as alternative feedstocks for poly-3-hydroxybutyrate (PHB) production as these potentially CO₂-derived molecules are naturally assimilated by Cupriavidus necator. Both organic acids were individually evaluated in fed-batch fermentations at bioreactor scale. Acetic acid was revealed as the most promising carbon source yielding 42.3 g L^-1 PHB, whereas no significant amount of PHB was produced from formic acid. Hence, acetic acid was further used as the substrate during process intensification. Key performance characteristics, including process stability, PHB titer, and productivity were optimized by introducing NH₄-acetate as the nitrogen source, extending the growth phase, and implementing a repeated fed-batch procedure, respectively. These advanced fermentation strategies resulted in the establishment of a stable fermentation process reaching 58.5 g L^-1 PHB, while doubling the productivity to 0.93 g L^-1 h^-1 PHB.

CO₂-based poly(3-hydroxybutyrate) (PHB) can be produced by the versatile bacterium Cupriavidus necator through chemolithoautotrophic fermentation, using a gas mixture consisting of CO₂, H₂, and O₂. Despite offering a propitious route for carbon-neutral bioplastic manufacturing, its adoption is currently hampered by the wide explosive range of the required gas mixture, as well as the limited gas-to-liquid mass transfer rates. To address these challenges, pressure fermentation was applied as a robust and effective strategy, while ensuring safe operation by adhering to the limiting O₂ concentration, utilizing state-of-the-art bioreactors. Consequently, exponential growth could be prolonged, boosting CO₂-based PHB production from 10.8 g/L at 1.5 bar up to 29.6 g/L at 3 bar. The production gain closely aligns with the theoretical calculations, except for when the pressure was increased up to 4 bar. Overall, the demonstrated increase in PHB production underscores the potential of pressure fermentation to enhance aerobic gas fermentation.

Gas fermentation is gaining attention as a crucial technology for converting gaseous feedstocks into value-added chemicals. Despite numerous efforts over the past decade to investigate these innovative processes at a lab scale, to date, the evaluation of the technologies in relevant industrial environments is scarce. This study examines the fermentative production of acetate from biomass-derived syngas using Moorella thermoacetica. A mobile gas fermentation pilot plant was coupled to a bubbling fluidized-bed gasifier with syngas purification to convert crushed bark-derived syngas. The syngas purification steps included hot filtration, catalytic reforming, and final syngas cleaning. Different latter configurations were evaluated to enable a simplified syngas cleaning configuration for microbial syngas conversion compared to conventional catalytic synthesis. Fermentation tests using ultra-cleaned syngas showed comparable microbial growth (1.3 g/L) and acetate production (22.3 g/L) to the benchmark fermentation of synthetic gases (1.2 g/L of biomass and 25.2 g/L of acetate). Additional fermentation trials on partially purified syngas streams identified H₂S and HCN as the primary inhibitory compounds. They also indicated that caustic scrubbing is an adequate and simplified final gas cleaning step to facilitate extended microbial fermentation. Overall, this study shows the potential of gas fermentation to valorize crude gaseous feedstocks, such as industrial off-gases, into platform chemicals.

Poly(3-hydroxybutyrate) (PHB) is a microbially produced biopolymer that is emerging as a propitious alternative to petroleum-based plastics owing to its biodegradable and biocompatible properties. However, to date, the relatively high costs related to the PHB production process are hampering its widespread commercialization. Since feedstock costs add up to half of the total production costs, ample research has been focusing on the use of inexpensive industrial side streams as carbon sources. While various industrial side streams such as second-generation carbohydrates, lignocellulose, lipids, and glycerol have been extensively investigated in liquid fermentation processes, also gaseous sources, including carbon dioxide, carbon monoxide, and methane, are gaining attention as substrates for gas fermentation. In addition, recent studies have investigated two-stage processes to convert waste gases into PHB via organic acids or alcohols. In this review, a variety of different industrial side streams are discussed as more sustainable and economical carbon sources for microbial PHB production. In particular, a comprehensive overview of recent developments and remaining challenges in fermentation strategies using these feedstocks is provided, considering technical, environmental, and economic aspects to shed light on their industrial feasibility. As such, this review aims to contribute to the global shift towards a zero-waste bio-economy and more sustainable materials.

Abundant biomass, including industrial waste streams and second-generation (2G) and third-generation (3G) feedstocks, offers significant potential for sustainable bioconversion, nevertheless challenges such as fermentation inhibitors, CO₂ losses and substrate selectivity of traditional microbial hosts hinder process efficiency. In this study, we address these challenges by exploring acetogenic bacteria as alternative microbial hosts. Using a newly established high-throughput method, acetogens were evaluated for their capacity to hydrolyse and metabolize variety of substrates derived from 2G and 3G feedstocks and industrial waste streams. Our findings demonstrate metabolic versatility of acetogens in converting biomass-derived substrates into a wide array of products while also exhibiting resilience to common fermentation inhibitors. These unique capabilities position acetogens as promising alternatives that could potentially outperform conventional production hosts in achieving 100% biomass valorization while underscoring the need for further research into critical areas, such as the utilization of mixed substrates under industrially relevant conditions.