Over the last decade, a significant amount of research has been done on sphingosine-1-phosphate (S1P). S1P regulates survival, proliferation and migration of breast cancer cells45, as well as playing an important but still unknown role in ovarian46, prostate, and glioma47 cell carcinomas. S1P plays a significant role in the regulation, proliferation and angiogenesis, and is therefore an area of considerable interest for cancer research. Previous research in our group has shown that two stilbenoid sphingosine analogs (1 & 2) were effective at sphingosine kinase (SphK) inhibition in addition to showing down-regulation of Vascular Endothelial Growth Factor (VEGF), a novel and distinctly different mechanism of action compared to reported sphingosine kinase inhibitors. Both compounds showed novel toxicity toward the parasite Toxoplasma gondii. T. gondii causes toxoplasmosis; a disease carried by up to 1/3rd of the world’s population and one that has been implicated in schizophrenia. Unfortunately, use of 1 & 2 as fluorescent probes is limited; their emission wavelengths fall between 360 and 380 nm, within the range of background fluorescence. The synthesis of five fluorescent sphingosine analogs is described herein, three of which have emission wavelengths above 420 nm. These compounds show single digit µM cytotoxicity towards T. gondii. The second generation synthesis of Fingolimod (FTY720) is also described. The process was optimized for large-scale production. Improvements to the synthesis included increased atom economy, purification and overall reaction efficiency. Yield was increased to 36%, from 33% in the first generation synthesis. Lastly, a synthesis of p-divinylbenzene is described, using “green” solvents and easy sample work-up. The product contains no meta impurities, an issue that arises when p-divinylbenzene is purchased commercially. / Thesis / Candidate in Philosophy

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No description available.

近年発酵生産が注目されているコハク酸の抽出分離法についてイオン液体を含む液膜法及び水性２相抽出法を用いて研究した．イオン液体を含む含浸型液膜により，受容相に塩酸水溶液を用いることでコハク酸の上り坂輸送を達成できた．次にイオン液体を含む高分子溶媒膜によるコハク酸の透過を検討した．最適な条件は含浸液膜と異なり，透過にはコハク酸の各化学種が関与していた．最後に水溶性有機溶媒やイオン液体を用いた水性２相抽出法によるコハク酸の抽出を行った．塩および糖を相分離剤とすることで，発酵液で想定されるコハク酸の80％以上の抽出が達成できた． / Succinic acid is widely used in many industrial sectors; pharmacy, textile, food, etc. The bio-production of succinic acid from renewable resource was desired because the petrochemical production of succinic acid became costly. In this study, the extraction and separation techniques of succinic acid by liquid membranes (LMs) and aqueous two-phase system (ATPS) containing ionic liquids (ILs) were exploited. The permeation mechanisms and optical conditions were elucidated. / 博士(工学) / Doctor of Philosophy in Engineering / 同志社大学 / Doshisha University

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No description available.

Aqueous electrolyte solutions at temperature and pressure conditions near the critical point of water are difficult to describe using traditional equations of state based upon the excess Gibbs energy. Models based upon the residual Helmholtz energy have proven more effective. Anderko and Pitzer1 developed a residual Helmholtz energy model (AP) for aqueous electrolyte solutions in which the electrolyte is assumed to be fully associated. The model has been effectively used in describing densities and vapor-liquid equilibria for simple electrolyte systems. The model is less effective for describing enthalpic properties such as heats of dilution. Oscarson and coworkers2, 3 modified the AP model for NaCl solutions by adding a term accounting for the change in Helmholtz energy as a result of aqueous dissociation reactions. This new model, called the RI model, is more accurate than the AP model at conditions where the NaCl dissociates more fully into ions. Liu et. al4, 5 modified the RI model by adding a term to describe interactions between ions in solution and by regressing new model parameter values. This new model, called the RIII model, is more accurate than both the AP model and the RI model and may be used to predict species concentrations in solution as a result of aqueous phase reactions. The RIII model has substantial thermodynamic inconsistencies, however, and is poorly suited for describing mixed solute solutions. This dissertation presents the RIV model which is an electrolyte solution model for solutions in the ranges of 350 °C to 400 °C and 18 MPa to 40 MPa. The RIV model has been applied to aqueous NaCl solutions and aqueous LiCl solutions. The RIV model is a modification of the AP model and includes aqueous phase reactions implicitly through fundamental species interactions. The RIV model is thermodynamically consistent. It is capable of describing densities and heats of dilution. Density predictions from the RIV model are less accurate than the AP model predictions (6.66 % error vs. 3.51 % error) but are reasonable. The heats of dilution predictions from the RIV model are much more accurate than those from the AP model (25.16 % error vs. 78.78 % error). Predictions of the ionic species concentration from the RIV model are likely to be poor as indicated by the poor agreement between experimental values and calculated values of equilibrium constants valid at infinite dilution. In order to provide the necessary data from which to regress the parameters of the RIV model, experimental heat of dilution values were determined using flow calorimetry techniques. These values are also reported in this dissertation.

The food industry is increasingly concerned with operational sustainability and food waste reduction. In the United States, the tomato industry was worth $1 billion in 2020, and tomatoes are currently ranked second as the most consumed vegetable after potatoes. Tomato processors have striven to valorize pomace by-products, which contain seeds with valuable compounds such as 45% fiber, 30% oil, and 26% protein. The U.S. peach industry, which was worth $599 million in 2017, is also looking for alternative ways to utilize their seeds, made up of 52% oil. Both tomato and peach seeds are rich in bioactive compounds such as carotenoids and polyphenols, respectively. Meanwhile, global edible oil production is forecasted to reach 632 million tons in 2022, and there is increasing interest to produce specialty oils. Organic solvent extractions are commonly used to extract oils from various commodities in the food industry, but this method comes with some environmental concerns, such as toxicity and flammability. Enzyme-assisted aqueous extractions (EAEP) have been proposed as a green alternative to solvent extractions of oilseeds. However, research on the economic feasibility of this process has been limited. There is a need for a better understanding of the potential of EAEP and performing such analysis on peach and tomato seeds would be valuable based on the importance of these commodities in California and in the U.S. in general. To determine whether applying such a process to tomato and peach seeds would be valuable, data on oil yields from EAEP, quality of the extracted oils, and economics associated with EAEP is needed. The U.S. snack food industry was worth approximately $42 billion as of 2019, and this market continues to grow as an increasing number of people eat snacks as meal replacements. There is also growing consumer demand for natural and functional foods, which offer health benefits beyond basic nutritional value, such as reducing risk of disease. Therefore, this consumer landscape provides excellent opportunities for by-products from vegetable and fruit processing to be transformed into functional ingredients. With these key ideas in mind, the objectives of this research were 1) to determine the impact of various extraction factors on oil yields from EAEP of tomato and peach seeds; 2) to evaluate the effect of aqueous extractions on oil quality; 3) study whether these processing steps are economically feasible for industrial commercialization and 4) identify another potential use for tomato pomace as a functional ingredient in snack food application. Tomato and peach seeds were each isolated and ground into a flour, then analyzed for fiber, fat, protein, moisture, and ash content. The effects of pH (set to 3, 9, and sequential adjustment of 3 followed by 9), time (2–8 h), and addition of cellulase, protease, and 1:1 enzymatic cocktail at a concentration of 4% were evaluated during aqueous extractions of tomato oils. Peach oil extractions were evaluated for the same parameters except for time which was fixed at 2 h. The impact of the pH of aqueous extractions on the oxidative stability and nutritional composition of tomato and peach oils were also determined. Techno-economic analyses were conducted using Superpro software to estimate operational costs and profits from this process. To assess the value of tomato by-product as a functional ingredient, pomace flour was added to crisp snacks at usage levels of 0 and 7% (w/w flour basis). Crisp snacks were stored at ambient room temperature and frozen temperatures (-23 °C) for ten weeks. Proximate analyses on the crisp snacks were conducted. Sensory evaluation was performed over time using hedonic scoring surveys. For tomato seeds, the highest oil yield (41%) was obtained during 2 h extractions at pH 9, which were 68% higher than from 2 h extractions performed at pH 3. Enzyme addition was only beneficial during 8 h extractions of tomato seeds using 4% cellulase at pH 3, which led to 53% higher oil yield compared to the control performed in the same conditions without enzymes. Increasing incubation time from 2 to 8 h improved oil yields by 63% for cellulase extractions at pH 3 and 69% for protease extractions at pH 9. Peach oil yield of the controls at pH 9 (53%) and the yield obtained with protease extractions at pH 9 (45%) were approximately eight times higher than all extractions set to pH 3. Once extracted, some properties of the tomato and peach oils were determined. The pH of the aqueous extractions did not have a significant effect on quality parameters on both tomato and peach oils, except for polyphenol content, DPPH production, peroxide and TBARS values. Overall, both tomato and peach oils had peroxide and free fatty acid values comparable to other specialty oils, such as sunflower and sesame oils. From a techno-economic point of view, tomato oil production at flow rate of 1,752,000 kg pomace/yr garnered $13 million in profit (equivalent to $7.42/kg pomace) after a payback time of 1.6 years, which was more profitable than the industry’s current pomace disposal practice. Peach oil processing became profitable once the production scale reached 65,700,000 kg pits/yr, which led to $15.5 million in profit ($0.24/kg pits). Overall, production of tomato oil was more profitable than peach oil. When tomato pomace was added into crisp snacks at 7% concentration, fiber increased by 35.2% compared to the control. Moisture content and water activity for the control and pomace crisp snacks remained constant during room temperature and frozen storage for ten weeks. However, the addition of tomato pomace led to a bitter aftertaste of the crisp snacks and reduced overall liking scores by panelists compared to scores for the control crisp snacks. Overall, this research took multiple, innovative approaches to valorize tomato and peach seeds, while also studying the economic, environmental, and industrial implications of such approaches. It highlighted alternative, sustainable strategies of how tomato and peach by-products could be repurposed to reduce waste and make value-added food products.

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Elastin-like polypeptides (ELP) are artificially designed protein biopolymers that can be produced by living organisms. These proteins have the unique ability to undergo reversible inverse phase transition, in response to changes in temperature and/or addition of chaotropic salts. Below the transition temperature (T1) , ELP is soluble in water. Increasing the temperature above Ti, ELP coacervates into an aqeous ELP-rich phase. In this thesis, this unique feature of ELP was used in for recombinant protein purification and for the formation of aqueous multiple-phase systems. For protein purification, ELP was fused with an intein and a model protein (thioredoxin), to demonstrate a simple and inexpensive approach for recombinant protein purification. The ELP tags replace the chromatographic media and the intein replaces the use of the protease in conventional methods. Using ELP tags was found to be consistent with large -scale recombinant protein production/purification by purifying an ELP tagged protein using a stirred cell equipped with a microfiltration membrane. When the temperature and/or salt concer.tration is increased for mixtures containing free ELP and ELP tagged proteins, simultaneous phase transition takes place. This served as the basis for the development of a method suitable for selectively recovering molecules from complex mixtures with high specificity, full reversibility, and virtually unlimited affinity. The second parts of this thesis focus on the ability of ELP to form aqueous twophase systems (A TPS) in vitro and most importantly, in vivo- with the formation of aqueous microcompartments in living cells. These compartments exclude the protein making machinery of the cell, acting as depots for newly expressed protein. It is also shown (in vitro) that ELP bastd droplets exclude proteases, protecting proteins from degradation. These observations are important for high-level production of recombinant proteins. Also described, is the formation of protein based aqueous multiphasic systems, with tunable morphologies. / Thesis / Doctor of Philosophy (PhD)