Engineering Commons^™

Exploring Versatile Glycosyltransferases From Microorganisms For Natural Product Biosynthesis, Jie Ren

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This dissertation is mainly focused on exploring versatile glycosyltransferases from different microorganisms with the purpose of biosynthesizing bioactive natural product glycosides through microbial fermentation. This engineered biosynthesis approach is environmentally friendly, safe, and efficient compared with plant extraction, chemical synthesis, and enzymatic synthesis. Additionally, we applied these characterized glycosyltransferases as efficient biocatalytic tools to produce novel glycosides to expand the chemical repertoire of nature. These novel compounds showed improved water solubility and some of them exhibited enhanced stability and biological activity, which laid the foundation for the development of new therapeutic agents.

In my dissertation research, I have discovered two …

Surface-Functionalized Silica Nanocarriers For Mitigating Water Stress In Wheat And Benefiting The Root Microbiome, Anthony Cartwright

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Changes in climate and shifting patterns of drought threaten the growth of important cash crops like wheat. The element silicon serves as a plant nutrient and shows promise for strengthening wheat against drought while remaining safe to both the crop and the positive bacteria that grow on its roots. Silicon can be added to wheat in the form of silicon-dioxide nanoparticles featuring protective coatings made from plant-beneficial nutrients. These nanoparticles can be engineered with high surface area or porous structures allowing them to be loaded with additional nutrients that can be delivered to crops. In a laboratory setting, such nanoparticles …

Increasing Production Of Therapeutic Mabs In Cho Cells Through Genetic Engineering, Charles Barentine

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Between 2014 and 2018, the global market for therapeutic monoclonal antibodies (mAbs) rose from $60 billion to $115.2 billion with a projected value of $300 billion by 2025. These molecules are used to effectively treat some of the most challenging illnesses from auto-immune diseases to cancer. While mAbs are highly valuable with potent applications, their production at scale remains an outstanding challenge. These molecules are largely produced in Chinese Hamster Ovary (CHO) cells that require highly specific conditions to produce a useful product.

Genetic engineering presents one solution to overcome productivity limits. With the advent of CRISPR (clustered regularly interspaced …

Low Resource Assay For Tracking Sars-Cov-2 In Wastewater, Julissa Van Renselaar

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Wastewater based epidemiology (WBE) allows for the tracking of nucleic acid of SARS-CoV-2 in wastewater which gives the opportunity for the public and government officials to be informed about the infectivity of the virus in a community. Advances have been made in WBE that have allowed for higher performance, lower resource use, and faster turnaround time. An adapted concentration method of spin column direct extraction has yielded a proxy virus recovery of 83%, consumable cost of $2.01 per sample, and a turnaround time of 0.33 hour/sample. Other concentration methods have lower proxy virus recoveries, comparable cost, and comparable turnaround time. …

Upstream Methods For Enhancing Engineered Curcumin Biosynthesis, Caleb D. Barton

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Curcumin is a bright orange compound with myriad applications for human health and wellness. Curcumin occurs naturally in the plant Curcuma longa (commonly known as turmeric) but must be extracted from the roots in an environmentally unfriendly fashion to obtain commercially relevant amounts of the compound. In addition, extraction of curcumin from turmeric spice yields a mixture of various curcuminoids, presenting an issue for isolating it in its pure form and complicating its use in clinical settings.

Heterologous biosynthetic production of curcumin in Escherichia coli has been used extensively as a viable alternative to plant extraction but suffers from poor …

Biomethanation And Alkaline Wet Air Oxidation Of Water Hyacinth (Pontederia Crassipes) From Ozama River, Dominican Republic, Yessica A. Castro

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Obtaining valuable products from environmental remediation waste is a sustainable approach that contributes to the ecological well-being of developing countries. In the present work, the feasibility of the water hyacinth anaerobic digestion as a post-weed management practice in the Ozama River (Dominican Republic) was demonstrated. The estimated energy required for harvesting was lower than that produced during digestion. The biomethanation of water hyacinth was improved by Alkaline Wet Air Oxidation (AWAO), a thermochemical pretreatment process that almost doubled the methane production rate and increased the yield by 24% when conducted at high temperatures. At lower temperatures, the methane yield of …

Cytomegalovirus Inhibition By Pluronic-Encapsulated Quercetin And Synergy With Ganciclovir, Ian Wadsworth

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Congenital cytomegalovirus (CMV) is the leading viral cause of birth defects worldwide and leads to a variety of complications. Ganciclovir is the most common anti-CMV drug used but often causes severe side effects. Quercetin, a plant pigment, shows anti-CMV activity that may provide a less toxic alternative to ganciclovir, or when co-administered, reduce ganciclovir treatments. However, a drug carrier is needed to deliver quercetin for CMV treatment. In this study, three Pluronic polymers (P123, F127, and F68) were investigated as a quercetin drug delivery system (DDS) for CMV treatment.

All selected Pluronics successfully encapsulated quercetin at varying degrees of efficacy. …

Characterization Of Chinese Hamster Ovary Cell Performance With Pentanoic Acid And N-Acetyl Cysteine Treatment, Joseph Camire

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

To improve the cost of producing protein therapeutics manufacturers can attempt to increase the amount of protein produced per unit volume. Some small fatty-acids have shown the ability to increase protein concentrations in some manufacturing systems. An example system is in the industrially important cell line derived from a Chinese Hamster Ovary (CHO). A drawback of adding these fatty-acids is they can increase the rate of cell death.

A potential way to minimize the cell death rate, caused by adding fatty-acids, is to also add additional anti-oxidants. Through a series of studies, we demonstrate production with different fatty-acids, butanoic acid …

Protecting Crops From Abiotic Stress: Copper Oxide Nanoparticle Effects On Wheat And A Beneficial Rhizobacterium, Matthew Potter

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Nanoparticles (NPs) are defined as particles less than 100 nm in at least one dimension. CuO NPs have possible applications in agriculture as micronutrient sources, pesticides, and enhancers of crop stress tolerance. Here, three aspects of CuO NP agricultural applications are studied: 1) the potential of CuO NPs to prevent wheat lodging-when crops irreversibly fall over; 2) CuO NP-induced drought tolerance in wheat seedlings; and 3) the effects of CuO NPs on outer membrane vesicle (OMV) production by Pseudomonas chlororaphis O6 (PcO6), a plant-health promoting bacterium.

Wheat grown 7 d exposed to CuO NPs in the growth matrix …

Determination Of Gas Emission Characteristics From Animal Wastes Using A Multiplexed Portable Ftir-Surface Chamber System, Pakorn Sutitarnnontr

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Livestock production is a growing source of air pollution at regional, national, and global scales. Improved livestock manure management has the potential to reduce environmental impacts; however, there is an urgent need for cost-efficient, reliable, and easy to maintain measurement and monitoring capabilities to precisely quantify emissions from livestock manure. This research describes and evaluates a novel measurement method based on the multiplexed portable Fourier Transform Infrared (FTIR) spectroscopy analyzer - surface chamber techniques for continuous measurements and monitoring gas emissions from manure sources. The multiplexing system was designed and developed to automate the chamber network, controlling the movement of …

Observation Of Struvite In The Mixed Microalgae Biofilm Matrix Of A Rotating Algal Biofilm Reactor During Nutrient Removal From Municipal Anaerobic Digester Filtrate, Kyle M. Hillman

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Central Valley Water Reclamation Facility (CVWRF) in Salt Lake City is the largest municipal wastewater treatment plant in Utah and must meet new and rigorous nutrient effluent standards–over 95% reduction in phosphorus output by 2025. Filtrate from CVWRF anaerobic digesters contains high levels of nitrogen, phosphorus, and magnesium. Supersaturation of these constituents leads to nuisance struvite precipitation that clogs belts, pumps, and pipes downstream of anaerobic digesters. Struvite is a mineral precipitate composed of equimolar magnesium, ammonium, and phosphate. Controlled precipitation of struvite helps prevent clogging and scaling, removes phosphate from wastewater, and generates a marketable fertilizer product.

Struvite was …

Expression Systems For Synthetic Spider Silk Protein Production, Michaela R. Hugie

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silk is a biodegradable and biocompatible natural material that is stronger than steel and more elastic than nylon. These properties make spider silk a desirable material for many commercial products, ranging from textiles to biomedical materials. Due to spiders’ cannibalistic and territorial nature it is impossible to farm them to produce spider silk at a high enough yield to meet product demands. Therefore, a bioengineered synthetic process is necessary to produce spider silk. Synthetic spider silk has been produced in bacteria, goats, yeast, plants, mammalian cells and silkworms, but none of these processes provided a commercially viable yield or …

Spiderworms: Using Silkworms As Hosts To Produce A Hybrid Silkworm-Spider Silk Fiber, Ana Laura Licon

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silk has received significant attention due to its fascinating mechanical properties. Given the solitary and cannibalistic behavior of spiders, spider silk farming is impractical. Unlike spiders, silkworms are capable of producing large quantities of a fibrous product in a manner mimetic to spiders, and there already exists an industry to process cocoons into threads and textiles for many applications. The combination of silk farming (sericulture), a millennia old practice, and modern advancements in genetic engineering has given rise to an innovative biomaterial inspired by nature; transgenic silkworm silk.

This project focuses on the creation of chimeric silkworm-spider silk fibers …

Optimization Of Biogas Production By Use Of A Microbially Enhanced Inoculum, Anna Doloman

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A renewable energy source, biogas, comprises of methane (80%) and carbon dioxide (15%), and is a great alternative to the conventional fossil-based fuels, such as coal, gas and oil. Biogas is created during anaerobic biological digestion of waste materials, such as landfill material, animal manure, wastewater, algal biomass, industrial organic waste etc. A biogas potential from organic waste in the United States is estimated at about 9 million tons per year and technology allows capture of greenhouse gases, such as methane and carbon dioxide, into a form of a fuel. In the light of global climate change and efforts to …

Characterization Of Biofilms In A Synthetic Rhizosphere Using Hollow Fiber Root-Mimetic Systems, Michelle Bonebrake

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The area around a plant’s roots hosts a complex and diverse microbial community. This environment can include a large number of bacteria that live on the surface of the root and benefit from the nutrients that the roots exude into the soil. These microbes can in turn be beneficial to the plant by protecting the roots from harmful fungi or stressful environmental conditions such as drought. In this thesis, several root-mimetic systems (RMSs) were developed for the study and growth of plant-beneficial bacteria in the laboratory environment. The RMS uses a porous hollow fiber used in hemodialysis as a surface …

Exploring The Capacity Of Bacteria For Natural Product Biosynthesis, Ozkan Fidan

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This dissertation is focused on exploring the potential of bacteria for the biosynthesis of natural products with the purposes of generating novel natural product derivatives and of improving the titer of pharmaceutically important natural products.

A wide variety of compounds from various sources have been historically used in the treatment and prevention of diseases. Natural products as a major source of new drugs are extensively explored due to their huge structural diversity and promising biological activities such as antimicrobial, anticancer, antifungal, antiviral and antioxidant properties. For instance, penicillin as an early-discovered antimicrobial agent has saved millions of lives, indicating the …

Hydrodeoxygenation Of Pinyon-Juniper Catalytic Pyrolysis Oil, Hossein Jahromi

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Catalytic hydrodeoxygenation (HDO), is an effective process to convert oxygenated compounds to hydrocarbons. This process is widely used for improving the negative properties of biomass-derived pyrolysis oils (bio-oils) such as high acidity, poor stability, and low heating value. During this process oxygen is removed from the bio-oil in the form of water, thus the liquid product of HDO process consists of aqueous phase and hydrocarbon phase that can be easily separated. Synthesis of efficient HDO catalyst has been a major challenge in the field of bio-oil upgrading. Red mud, which is an alkaline waste from alumina industry was used to …

In Vitro Simulation Of Microgravity Induced Muscle Loss Successfully Increases Expression Of Key In Vivo Atrophy Markers, Charles P. Harding

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Muscle loss from lack of activity is a serious issue for immobilized patients on Earth and in human spaceflight, where the low gravity environment prevents normal muscle activity. Simulating muscle loss in cultured cells is an important step in understanding how this condition occurs. This work evaluates different means of simulating muscle loss and selects the one that most closely mimics the cellular responses seen in animals and humans.

To simulate the microgravity environment of spaceflight, mouse skeletal muscle cells were grown in a rotary cell culture system (RCCS). Growing the cells within a natural gelled substrate was compared against …

Production And Biocompatibility Of Spider Silk Proteins In Goat Milk, Richard E. Decker Jr

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Due to its strength, flexibility, and biocompatibility, spider silk is a highly appealing material for applications in the medical field. Unfortunately, natural spider silk is difficult to obtain in large quantities because spiders are territorial and cannibalistic, making them impractical to farm. Synthetic spider silk proteins produced by transgenic hosts such as bacteria and goats have made it possible to obtain the quantities of spider silk needed to study it more fully and to investigate its potential uses. The spider silk proteins produced in our laboratory do not have an optimal purification method to remove all of the non-biocompatible contaminants …

Identification And Engineering Of Nonribosomal Peptide Biosynthetic Systems, Fuchao Xu

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This research focuses on the understanding and engineering of nonribosomal peptide biosynthetic pathways in Streptomyces coelicolor CH999, Escherichia coli BAP1 and Saccharomyces cerevisiae BJ5464-NpgA. The biosynthetic systems of indigoidine from bacteria and beauvericin/bassianolide from fungi were studied in this research. The production of these valuble products was significantly increased by enhancing their synthetic pathway with metabolic engineering approaches.

Indigoidine is a bacterial natural product with antioxidant and antimicrobial activities. Its bright blue color resembles the industrial dye indigo, thus representing a new natural blue dye that may find uses in industry. Indigo is a dark blue crystalline powder and has …

Development And Characterization Of Aqueous-Based Recombinant Spider Silk Protein Biomaterials With Investigations Into Potential Applications, Thomas I. Harris

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silks are incredible natural materials that possess desirable combinations of strength, elasticity, weight, and robustness. Other properties such as biocompatibility and biodegradability further increase the worth of these materials. The possibility of farming spiders is impractical due to spiders’ natural behaviors. Modern biotechnologies have allowed for recombinant spider silk proteins (rSSps) to be produced without the use of spiders. However, the features responsible for spider silks impressive properties can cause difficulties with producing silk materials. A recently developed water-based and biomimetic solvation method has provided a solution to such difficulties and has also led to novel silk biomaterials. Most …

Biomass And Phycocyanin From Oil And Natural Gas Extraction Produced Water Utilizing A Cyanobacteria Dominated Rotating Algal Biofilm Reactor (Rabr), Jonathan L. Wood

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The production of cyanobacterial biofilm biomass and phycocyanin from Rotating Algal Biofilm Reactors utilizing undiluted produced water from oil and natural gas extraction as a culture medium was investigated in this study. Produced water is the largest waste stream generated by the oil and natural gas industries and represents a large volume of non-potable water that may be available for algae culture with minimal impact on freshwater resources. Combining the use of produced wastewater as culture medium with the production of high value algal pigments, such as phycocyanin, may increase the economic viability of algae culture and wastewater purification. High …

Development And Optimization Of A Produced Water, Biofilm Based Microalgae Cultivation System For Biocrude Conversion With Hydrothermal Liquefaction, Benjamin L. Peterson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Extraction of oil and gas in Utah’s Uintah Basin results in large quantities of wastewater, or produced water, with nutrients and residual organic chemical that represent a significant resource for producing energy-related and value-added products. Produced water was obtained as a biomass producing nutrient source from industries operating in Utah’s Uintah Basin. Within the Uintah Basin (defined as Uintah and Duchesne Counties within Utah) approximately 93 million barrels of water were produced in 2013 while only 11% of the water was disposed of through evaporation, with the national average at 2%. The rest is reinjected into the subsurface.

The goal …

Novel Methods To Produce Large Recombinant Spider Silk Proteins Via Polymerization, Nathan L. Hebert

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Spider silk has long been a subject of scientific research due to its remarkable mechanical properties. Until recently, there has been no way to effectively obtain spider silk except by harvesting it from individual spiders. With advances in technology, the genes that code for the individual spider silk proteins have been isolated and genetically engineered into other hosts to produce recombinant spider silk proteins (rSSp) of varying sizes, Larger rSSp have correspondingly greater mechanical properties in any resulting materials. Using current production methods, larger rSSp cannot be produced in commercially viable quantities while simultaneously being economically viable. The current production …

Silane Modulation Of Protein Conformation And Self-Assembly, Abul Bashar Mohammad Giasuddin

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This research focused on development of nanoparticle- based therapeutics against amyloid fibrils. Amyloid fibrils are associated with various diseases such as Parkinson’s, Huntington’s, mad cow disease, Alzheimer’s, and cataracts. Amyloid fibrils develop when proteins change their shape from a native form to a pathogenic “misfolded” form. The misfolded proteins have the ability to recruit more native proteins into the pathogenic forms, which self-assemble into amyloid fibrils that are hallmarks of the various protein-misfolding diseases listed above. Amyloid fibrils are highly resistant to degradation, which may contribute to the symptoms of amyloid diseases. Synthetic drugs, natural compounds, and antibodies are widely …

The Use Of Microfluidics And Dielectrophoresis For Separation, Concentration, And Identification Of Bacteria, Cynthia Hanson

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Typical bacterial analysis involves culturing and visualizing colonies on an array of agar plates. The growth patterns and colors among the array are used to identify the bacteria. For fast growing bacteria such as Escherichia coli, analysis will take one to two days. However, slow growing bacteria such as mycobacteria can take weeks to identify. In addition, there are some species of bacteria that are viable but nonculturable. This lengthy analysis time is unacceptable for life-threatening infections and emergency situations. It is clear that to decrease the analysis of the bacteria, the culturing and growth steps must be avoided. …

Investigations Of Polyhydroxyalkanoate Secretion And Production Using Sustainable Carbon Sources, Chad L. Nielsen

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Polyhydroxyalkanoates (PHAs) are a type of biologically-produced plastic known for their biocompatibility and biodegradability. They have the potential to replace petroleum-based plastics as an environmentally-friendly alternative. This is beneficial because the release of plastics into environments such as the ocean and the buildup of plastics in landfills are major concerns facing society today. Currently, however, PHAs are significantly more expensive than their petroleum-based counterparts. This is largely due to the cost of carbon sources and of extracting the bioplastics from bacteria. The goal of these studies was to examine replacing traditional carbon sources used in PHA production like sugar and …

Investigation And Engineering Of Polyketide Biosynthetic Pathways, Lei Sun

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This research is focused on investigation and engineering of natural product biosynthetic pathways for efficient production of pharmaceutically important molecules or generation of new bioactive molecules for drug development.

Natural products are an important source of therapeutics, such as chromomycin (anti-cancer), emodin (anti-inflammatory and anti-tumor) and sprolaxine (anti-Helicobacter pylori). Metabolic engineering of natural product biosynthetic pathways shows its promise for creating and producing valuable compounds with chemical diversity for drug discovery. One goal of this research is to create highly efficient strains to biosynthesize valuable natural products. The engineered Streptomyces roseiscleroticus strain constructed in this work showed higher …

Alternative Treatment Technologies For Low-Cost Industrial And Municipal Wastewater Management, Alan J. Hodges

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Roughly the same volume of water that rushes over the Niagara Falls is produced as wastewater in North America. This wastewater is treated through a variety of means to ensure that it can be safely returned to the natural ecosystem. This thesis examines two novel means for this treatment, one biological and one physical-chemical in nature, namely, Rotating Algae Biofilm Reactor treatment and expanded shale augmented coagulation-flocculation.

Rotating algae biofilm reactors (RABRs) support biofilm algae growth, and in turn, the algae take up harmful contaminants from the wastewater. This system was tested in wastewater from petroleum refining operations. The efficacy …

Algae-Based Biofilm Productivity And Treatment Of Dairy Wastewater: Effects Of Temperature And Organic Carbon Concentration, Zachary T. Fica

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Production of dairy and associated products is a source of millions of gallons of wastewater every year. Water used in cleaning feeding stalls as well as the liquid component of the animal waste are two of the major volumetric components of this wastewater. This water is nutrient rich, often limiting the viability as a land applied fertilizer. However, these same nutrients could be used as an inexpensive feedstock for the cultivation of algae, which can then be used to produce downstream products including animal feed and aquaculture.

As part of this study, algal biomass was cultivated on dairy wastewater from …