Esters are important solvents in multiple industries including adhesives, food, and pharmaceuticals. Although esters are biodegradable solvents, the conventional process of producing them is not eco-friendly because they are largely derived from petrochemicals. This has led scientists to consider implementing biological routes in their production process by incorporating heterologous or improving inherent esterification pathways. However, due to inequality in the biosynthesis of esters and their precursors (organic acid and alcohol), a significant amount of precursors are left unconverted, thereby lowering overall esterification efficiency. Therefore, the primary goal of the current research is to improve the ester titers by incorporating one more step of in vitro esterification with the culture broth, thereby esterifying the unconverted precursors using high efficiency commercial enzymes in the presence of compatible organic solvent. In principle, the medium containing the precursors will be treated with the enzyme in presence of organic solvent, where the precursors will be distributed in both the phases, aqueous and organic, based on their polarity, and the enzymatic esterification will happen at the interface. Hence, as a first step, efforts were made to optimize the reaction conditions, beginning with choosing the most efficient organic solvent and corresponding enzyme candidate. Our results showed that, for production of ethyl acetate through this reactive extraction approach, Novozyme435 exhibited significant esterification with chloroform, with almost 85% conversion efficiency. Further optimizations with phase ratios, pH and incubation time showed that the pH 6.0 (3.1 g/L) was the most optimum where ethyl acetate titer was found to improve 10 times than that at pH 7.0 (0.164 g/L) with the phase ratio of 1:1. The kinetic studies further added that the incubation at 37oC gives the maximum ethyl acetate production within 8h. After initial optimization studies, cell broth from E. coli cells transformed to overproduce an esterase was also tested with the reactive extraction method. It was found that there was a ~7.5X decrease in ethyl acetate production in the cell media versus synthetic samples with the same concentration of reactants. Such a large decrease indicates that enzymatic promiscuity or inhibition currently prevent the cell samples from reaching the same conversion as synthetic studies. To characterize the maximum reaction rate (Vmax) and affinity constants of the substrates to Novozym 435, further kinetic studies were performed with one minute of reaction. The mathematical model employed assumes that enzyme kinetics rather than diffusion was the rate limiting step, that the concentrations of reactants at the interface are equivalent to the initial concentration of reactants, and that neither substrate is an inhibitor. Vmax was found to be 18.5 Mmol min-1g-1 (of catalyst used), and the affinity constants were 0.957 M and 0.00557 M for acetic acid and ethanol respectively. Vmax was similar to literature values with Novozym 435, and the affinity constants indicate a much higher binding efficiency of ethanol in comparison to acetic acid, indicating that a cocktail of esters are likely produced from Novozym 435 in cell broth. Overall, moving away from fossil-fuel dependence is necessary to promote sustainable industry standards, and microbial cell factories combined with reactive extraction, if optimized for industrial applications, can replace harmful environmental procedures. By optimizing the reactive extraction process for ester production, biorefineries could become more competitive and economically feasible for numerous applications.

There has been a recent push to examine the materials that nature is able to synthesize and consider whether the materials that humans have invented are geomimetic in nature, and whether designing nature-inspired materials is economically and environmentally beneficial. Mesoporous silica represents a class of materials with pore sizes of 2-50 nm and has been studied in catalysis, separations, and drug delivery. It has generally been made using organosilicon precursors, but in this work, we demonstrate for the first time the successful synthesis of mesoporous silica with uniform mesoporosity of 10 nm using the mineral forsterite (Mg2SiO4) as a silica source, providing a potentially cheaper and more Earth-friendly route to making this technologically important material. Forsterite was synthesized by a solid-state chemistry route and underwent dissolution-reprecipitation in an aqueous acid solution containing the soft template surfactant, Pluronic P123. The formation of forsterite was confirmed with X-ray diffraction (XRD), the successful templating of surfactant was demonstrated with thermogravimetric analysis (TGA) and Fourier-transform infrared spectroscopy (FTIR), the surface area was determined through Brunauer-Emmett-Teller (BET) analysis, and pore size and distribution were demonstrated with Barrett-Joyner-Halenda (BJH) analysis. The synthesized mesoporous silica at optimal conditions has surface area of 740 m2/g and pore volume of 1.4 mL/g.

Realtime understanding of one’s complete metabolic state is crucial to controlling weight and managing chronic illnesses, such as diabetes. This project represents the development of a novel breath acetone sensor within the Biodesign Institute’s Center for Bioelectronics and Biosensors. The purpose is to determine if a sensor can be manufactured with the capacity to measure breath acetone concentrations typical of various levels of metabolic activity. For this purpose, a solution that selectively interacts with acetone was embedded in a sensor cartridge that is permeable to volatile organic compounds. After 30 minutes of exposure to a range of acetone concentrations, a color change response was observed in the sensors. Requiring only exposure to a breath, these novel sensor configurations may offer non-trivial improvements to clinical and at-home measurement of lipid metabolic rate.