Maria C. Cuellar

Microbial advanced biofuels production: overcoming emulsification challenges for large-scale operation

Isoprenoids and alkanes produced and secreted by microorganisms are emerging as an alternative biofuel for diesel and jet fuel replacements. In a similar way as for other bioprocesses comprising an organic liquid phase, the presence of microorganisms, medium composition, and process conditions may result in emulsion formation during fermentation, hindering product recovery. At the same time, a low-cost production process overcoming this challenge is required to make these advanced biofuels a feasible alternative. We review the main mechanisms and causes of emulsion formation during fermentation, because a better understanding on the microscale can give insights into how to improve large-scale processes and the process technology options that can address these challenges.

Microbial community-based polyhydroxyalkanoates (PHAs) production from wastewater: Techno-economic analysis and ex-ante environmental assessment.

This work investigates the potential for polyhydroxybutyrate (PHB) production from wastewater, from a techno-economic and an environmental perspective, examining scale-up opportunities and bottlenecks prior to commercialisation. Conceptual process design, economic, environmental impacts and sensitivity analysis are developed for one fermentation process and three downstream processing routes, based on alkali, surfactant-hypochlorite and solvent treatments. Environmentally and cost-wise, the alkali treatment is the most favourable with production costs of 1.40€/kg PHB, global warming potential of 2.4kgCO2-eq/kg PHB and non-renewable energy use of 106MJ/kg PHB. The solvent-based process yields the highest costs and environmental burdens: 1.95€/kg PHB, 4.30kgCO2-eq/kg PHB and 156MJ/kg PHB. The production of PHB from wastewater is identified as an interesting alternative to pure culture-polyhydroxyalkanoates production from sugars. However, these results are not yet competitive with those for the petrochemical counterparts. Additional performance improvements may be possible, through process integration and optimisation.

Recent advances in the microbial production and recovery of apolar molecules

Several apolar molecules of interest for the production of fuels and chemicals can nowadays be produced by fermentation. Those secreted from the microbial cell are of particular interest for large scale bioprocessing, since they allow for cell reuse, in situ product recovery and competitive production levels. So far, however, bioprocess strategies for fermentation and product recovery have been developed for addressing needs at the laboratory scale, rather than the process scale. Most commonly used strategies include extractive fermentations, product stripping in the gas phase, and off-line de-emulsification followed by intensive centrifugation. At the same time, current techno-economic studies at process scale have demonstrated the absolute need for significant improvements in both microorganism and process technology, for these processes to become competitive.

Breaking oil-in-water emulsions stabilized by yeast

Several biotechnological processes can show an undesirable formation of emulsions making difficult phase separation and product recovery. The breakup of oil-in-water emulsions stabilized by yeast was studied using different physical and chemical methods. These emulsions were composed by deionized water, hexadecane and commercial yeast (Saccharomyces cerevisiae). The stability of the emulsions was evaluated varying the yeast concentration from 7.47 to 22.11% (w/w) and the phases obtained after gravity separation were evaluated on chemical composition, droplet size distribution, rheological behavior and optical microscopy. The cream phase showed kinetic stability attributed to mechanisms as electrostatic repulsion between the droplets, a possible Pickering-type stabilization and the viscoelastic properties of the concentrated emulsion. Oil recovery from cream phase was performed using gravity separation, centrifugation, heating and addition of demulsifier agents (alcohols and magnetic nanoparticles). Long centrifugation time and high centrifugal forces (2 h/150,000×g) were necessary to obtain a complete oil recovery. The heat treatment (60°C) was not enough to promote a satisfactory oil separation. Addition of alcohols followed by centrifugation enhanced oil recovery: butanol addition allowed almost complete phase separation of the emulsion while ethanol addition resulted in 84% of oil recovery. Implementation of this method, however, would require additional steps for solvent separation. Addition of charged magnetic nanoparticles was effective by interacting electrostatically with the interface, resulting in emulsion destabilization under a magnetic field. This method reached almost 96% of oil recovery and it was potentially advantageous since no additional steps might be necessary for further purifying the recovered oil.

Fermentation broth components influence droplet coalescence and hinder advanced biofuel recovery during fermentation

Developments in synthetic biology enabled the microbial production of long chain hydrocarbons, which can be used as advanced biofuels in aviation or transportation. Currently, these fuels are not economically competitive due to their production costs. The current process offers room for improvement: by utilizing lignocellulosic feedstock, increasing microbial yields, and using cheaper process technology. Gravity separation is an example of the latter, for which droplet growth by coalescence is crucial. The aim of this study was to study the effect of fermentation broth components on droplet coalescence. Droplet coalescence was measured using two setups: a microfluidic chip and regular laboratory scale stirred vessel (2 L). Some fermentation broth components had a large impact on droplet coalescence. Especially components present in hydrolysed cellulosic biomass and mannoproteins from the yeast cell wall retard coalescence. To achieve a technically feasible gravity separation that can be integrated with the fermentation, the negative effects of these components on coalescence should be minimized. This could be achieved by redesign of the fermentation medium or adjusting the fermentation conditions, aiming to minimize the release of surface active components by the microorganisms. This way, another step can be made towards economically feasible advanced biofuel production.

Microbial Hydrocarbon Formation from Biomass

Fossil carbon sources mainly contain hydrocarbons, and these are used on a huge scale as fuel and chemicals. Producing hydrocarbons from biomass instead is receiving increased attention. Achievable yields are modest because oxygen atoms need to be removed from biomass, keeping only the lighter carbon and hydrogen atoms. Microorganisms can perform the required conversions, potentially with high selectivity, using metabolic pathways that often end with decarboxylation. Metabolic and protein engineering are used successfully to achieve hydrocarbon production levels that are relevant in a biorefinery context. This has led to pilot or demo processes for hydrocarbons such as isobutene, isoprene, and farnesene. In addition, some non-hydrocarbon fermentation products are being further converted into hydrocarbons using a final chemical step, for example, ethanol into ethene. The main advantage of direct microbial production of hydrocarbons, however, is their potentially easy recovery because they do not dissolve in fermentation broth.