College of Chemistry

Sunscreen has been used for thousands of years to protect skin from ultraviolet radiation. However, the use of modern commercial sunscreen containing oxybenzone, ZnO, and TiO₂ has raised concerns due to their negative effects on human health and the environment. In this study, we aim to establish an efficient microbial platform for production of shinorine, a UV light absorbing compound with anti-aging properties. First, we methodically selected an appropriate host for shinorine production by analyzing central carbon flux distribution data from prior studies alongside predictions from genome-scale metabolic models (GEMs). We enhanced shinorine productivity through CRISPRi-mediated downregulation and utilized shotgun proteomics to pinpoint potential competing pathways. Simultaneously, we improved the shinorine biosynthetic pathway by refining its design, optimizing promoter usage, and altering the strength of ribosome binding sites. Finally, we conducted amino acid feeding experiments under various conditions to identify the key limiting factors in shinorine production. The study combines meta-analysis of ¹³C-metabolic flux analysis, GEMs, synthetic biology, CRISPRi-mediated gene downregulation, and omics analysis to improve shinorine production, demonstrating the potential of Pseudomonas putida KT2440 as platform for shinorine production.

QS-21 is a potent vaccine adjuvant and remains the only saponin-based adjuvant that has been clinically approved for use in humans1,2. However, owing to the complex structure of QS-21, its availability is limited. Today, the supply depends on laborious extraction from the Chilean soapbark tree or on low-yielding total chemical synthesis3,4. Here we demonstrate the complete biosynthesis of QS-21 and its precursors, as well as structural derivatives, in engineered yeast strains. The successful biosynthesis in yeast requires fine-tuning of the hosts native pathway fluxes, as well as the functional and balanced expression of 38 heterologous enzymes. The required biosynthetic pathway spans seven enzyme families-a terpene synthase, P450s, nucleotide sugar synthases, glycosyltransferases, a coenzyme A ligase, acyl transferases and polyketide synthases-from six organisms, and mimics in yeast the subcellular compartmentalization of plants from the endoplasmic reticulum membrane to the cytosol. Finally, by taking advantage of the promiscuity of certain pathway enzymes, we produced structural analogues of QS-21 using this biosynthetic platform. This microbial production scheme will allow for the future establishment of a structure-activity relationship, and will thus enable the rational design of potent vaccine adjuvants.

Disordered rock salt oxides (DRX) have shown great promise as high-energy-density and sustainable Li-ion cathodes. While partial substitution of oxygen for fluorine in the rock salt framework has been related to increased capacity, lower charge-discharge hysteresis, and longer cycle life, fluorination is poorly characterized and controlled. This work presents a multistep method aimed at assessing fluorine incorporation into DRX cathodes, a challenging task due to the difficulty in distinguishing oxygen from fluorine using X-ray and neutron-based techniques and the presence of partially amorphous impurities in all DRX samples. This method is applied to Li1.25Mn0.25Ti0.5O1.75F0.25 prepared by solid-state synthesis and reveals that the presence of LiF impurities in the sample and F content in the DRX phase is well below the target. Those results are used for compositional optimization, and a synthesis product with drastically reduced LiF content and a DRX stoichiometry close to the new target composition (Li1.25Mn0.225Ti0.525O1.85F0.15) is obtained, demonstrating the effectiveness of the strategy. The analytical method is also applied to Li1.33Mn0.33Ti0.33O1.33F0.66 obtained via mechanochemical synthesis, and the results confirm that much higher fluorination levels can be achieved via ball-milling. Finally, a simple and rapid water washing procedure is developed to reduce the impurity content in as-prepared DRX samples: this procedure results in a ca. 10% increase in initial discharge capacity and a ca. 11% increase in capacity retention after 25 cycles for Li1.25Mn0.25Ti0.50O1.75F0.25. Overall, this work establishes new analytical and material processing methods that enable the development of more robust design rules for high-energy-density DRX cathodes.

Nanoparticle organic hybrid materials (NOHMs) have been proposed as excellent electrolytes for combined CO₂ capture and electrochemical conversion due to their conductive nature and chemical tunability. However, CO₂ capture behavior and transport properties of these electrolytes after CO₂ capture have not yet been studied. Here, we use a variety of nuclear magnetic resonance (NMR) techniques to explore the carbon speciation and transport properties of branched polyethylenimine (PEI) and PEI-grafted silica nanoparticles (denoted as NOHM-I-PEI) after CO₂ capture. Quantitative ¹³C NMR spectra collected at variable temperatures reveal that absorbed CO₂ exists as carbamates (RHNCOO^- or RR'NCOO^-) and carbonate/bicarbonate (CO₃^2-/HCO₃^-). The transport properties of PEI and NOHM-I-PEI studied using ¹H pulsed-field-gradient NMR, combined with molecular dynamics simulations, demonstrate that coulombic interactions between negatively and positively charged chains dominate in PEI, while the self-diffusion in NOHM-I-PEI is dominated by silica nanoparticles. These results provide strategies for selecting adsorbed forms of carbon for electrochemical reduction.

Understanding Li⁺ transport in organic-inorganic hybrid electrolytes, where Li⁺ has to lose its organic solvation shell to enter and transport through the inorganic phase, is crucial to the design of high-performance batteries. As a model system, we investigate a range of Li⁺-conducting particles suspended in a concentrated electrolyte. We show that large Li_1.3Al_0.3Ti_1.7P₃O₁₂ and Li₆PS₅Cl particles can enhance the overall conductivity of the electrolyte. When studying impedance using a cell with a large cell constant, the Nyquist plot shows two semicircles: a high-frequency semicircle related to ion transport in the bulk of both phases and a medium-frequency semicircle attributed to Li⁺ transporting through the particle/liquid interfaces. Contrary to the high-frequency resistance, the medium-frequency resistance increases with particle content and shows a higher activation energy. Furthermore, we show that small particles, requiring Li⁺ to overcome particle/liquid interfaces more frequently, are less effective in facilitating Li⁺ transport. Overall, this study provides a straightforward approach to study the Li⁺ transport behavior in hybrid electrolytes.

Moving toward a circular plastics economy is a vital aspect of global resource management. Chemical recycling of plastics ensures that high-value monomers can be recovered from depolymerized plastic waste, thus enabling circular manufacturing. However, to increase chemical recycling throughput in materials recovery facilities, the present understanding of polymer transport, diffusion, swelling, and heterogeneous deconstruction kinetics must be systematized to allow industrial-scale process design, spanning molecular to macroscopic regimes. To develop a framework for designing depolymerization processes, we examined acidolysis of circular polydiketoenamine elastomers. We used magnetic resonance to monitor spatially resolved observables in situ and then evaluated these data with a fractal method that treats nonlinear depolymerization kinetics. This approach delineated the roles played by network architecture and reaction medium on depolymerization outcomes, yielding parameters that facilitate comparisons between bulk processes. These streamlined methods to investigate polymer hydrolysis kinetics portend a general strategy for implementing chemical recycling on an industrial scale.

Isolated platinum(II) ions anchored at acid sites in the pores of zeolite HZSM-5, initially introduced by aqueous ion exchange, were reduced to form platinum nanoparticles that are stably dispersed with a narrow size distribution (1.3 ± 0.4 nm in average diameter). The nanoparticles were confined in reservoirs within the porous zeolite particles, as shown by electron beam tomography and the shape-selective catalysis of alkene hydrogenation. When the nanoparticles were oxidatively fragmented in dry air at elevated temperature, platinum returned to its initial in-pore atomically dispersed state with a charge of +2, as shown previously by X-ray absorption spectroscopy. The results determine the conditions under which platinum is retained within the pores of HZSM-5 particles during redox cycles that are characteristic of the reductive conditions of catalyst operation and the oxidative conditions of catalyst regeneration.

Perfluorosulfonic acid (PFSA) ionomers serve a vital role in the performance and stability of fuel-cell catalyst layers. These properties, in turn, depend on the colloidal processing of precursor inks. To understand the colloidal structure of fuel-cell catalyst layers, we explore the aggregation of PFSA ionomers dissolved in water/alcohol solutions and relate the predicted aggregation to experimental measurements of solution pH. Not all side chains contribute to measured pH because of burying inside particle aggregates. To account for the measured degree of dissociation, a new description is developed for how PFSA aggregates interact with each other. The developed single-counterion electrostatic repulsive pair potential from Part I is incorporated into the Smoluchowski collision-based kinetics of interacting aggregates with buried side chains. We demonstrate that the surrounding solvent mixture affects the degree of aggregation as well as the pH of the system primarily through the solution dielectric permittivity, which drives the strength of the interparticle repulsive energies. Successful pH prediction of Nafion ionomer dispersions in water/n-propanol solutions validates the numerical calculations. Nafion-dispersion pH measurements serve as a surrogate for Nafion particle-size distributions. The model and framework can be leveraged to explore different ink formulations.

Charged colloidal particles neutralized by a single counterion are increasingly important for many emerging technologies. Attention here is paid specifically to hydrogen fuel cells and water electrolyzers whose catalyst layers are manufactured from a perfluorinated sulfonic acid polymer (PFSA) suspended in aqueous/alcohol solutions. Partially dissolved PFSA aggregates, known collectively as ionomers, are stabilized by the electrostatic repulsion of overlapping diffuse double layers consisting of only protons dissociated from the suspended polymer. We denote such double layers containing no added electrolyte as "single ion". Size-distribution predictions build upon interparticle interaction potential energies from the Derjaguin-Landau-Verwey-Overbeek (DLVO) formalism. However, when only a single counterion is present in solution, classical DLVO electrostatic potential energies no longer apply. Accordingly, here a new formulation is proposed to describe how single-counterion diffuse double layers interact in colloidal suspensions. Part II (Srivastav, H.; Weber, A. Z.; Radke, C. J. Langmuir 2024 DOI: 10.1021/acs.langmuir.3c03904) of this contribution uses the new single-ion interaction energies to predict aggregated size distributions and the resulting solution pH of PFSA in mixtures of n-propanol and water. A single-counterion diffuse layer cannot reach an electrically neutral concentration far from a charged particle. Consequently, nowhere in the dispersion is the solvent neutral, and the diffuse layer emanating from one particle always experiences the presence of other particles (or walls). Thus, in addition to an intervening interparticle repulsive force, a backside osmotic force is always present. With this new construction, we establish that single-ion repulsive pair interaction energies are much larger than those of classical DLVO electrostatic potentials. The proposed single-ion electrostatic pair potential governs dramatic new dispersion behavior, including dispersions that are stable at a low volume fraction but unstable at a high volume fraction and finite volume-fraction dispersions that are unstable with fine particles but stable with coarse particles. The proposed single-counterion electrostatic pair potential provides a general expression for predicting colloidal behavior for any charged particle dispersion in ionizing solvents with no added electrolyte.

The evolution of pathway enzymes enhances the biosynthesis of high-value chemicals, crucial for pharmaceutical, and agrochemical applications. However, unpredictable evolutionary landscapes of pathway genes often hinder successful evolution. Here, the presence of complex epistasis is identifued within the representative naringenin biosynthetic pathway enzymes, hampering straightforward directed evolution. Subsequently, a biofoundry-assisted strategy is developed for pathway bottlenecking and debottlenecking, enabling the parallel evolution of all pathway enzymes along a predictable evolutionary trajectory in six weeks. This study then utilizes a machine learning model, ProEnsemble, to further balance the pathway by optimizing the transcription of individual genes. The broad applicability of this strategy is demonstrated by constructing an Escherichia coli chassis with evolved and balanced pathway genes, resulting in 3.65 g L-1 naringenin. The optimized naringenin chassis also demonstrates enhanced production of other flavonoids. This approach can be readily adapted for any given number of enzymes in the specific metabolic pathway, paving the way for automated chassis construction in contemporary biofoundries.