Matching Items (13)
Description
Synechocystis sp PCC 6803 is a photosynthetic cyanobacterium that can be easily transformed to produce molecules of interest; this has increased Synechocystis’ popularity as a clean energy platform. Synechocystis has been shown to produce and excrete molecules such as fatty acids, isoprene, etc. after appropriate genetic modification. Challenges faced for large–scale growth of modified Synechocystis include abiotic stress, microbial contamination and high processing costs of product and cell material. Research reported in this dissertation contributes to solutions to these challenges. First, abiotic stress was addressed by overexpression of the heat shock protein ClpB1. In contrast to the wild type, the ClpB1 overexpression mutant (Slr1641+) tolerated rapid temperature changes, but no difference was found between the strains when temperature shifts were slower. Combination of ClpB1 overexpression with DnaK2 overexpression (Slr1641+/Sll0170+) further increased thermotolerance. Next, we used a Synechocystis strain that carries an introduced isoprene synthase gene (IspS+) and that therefore produces isoprene. We attempted to increase isoprene yields by overexpression of key enzymes in the methyl erythritol phosphate (MEP) pathway that leads to synthesis of the isoprene precursor. Isoprene production was not increased greatly by MEP pathway induction, likely because of limitations in the affinity of the isoprene synthase for the substrate. Finally, two extraction principles, two–phase liquid extraction (e.g., with an organic and aqueous phase) and solid–liquid extraction (e.g., with a resin) were tested. Two–phase liquid extraction is suitable for separating isoprene but not fatty acids from the culture medium. Fatty acid removal required acidification or surfactant addition, which affected biocompatibility. Therefore, improvements of both the organism and product–harvesting methods can contribute to enhancing the potential of cyanobacteria as solar–powered biocatalysts for the production of petroleum substitutes.
ContributorsGonzalez Esquer, Cesar Raul (Author) / Vermaas, Willem (Thesis advisor) / Chandler, Douglas (Committee member) / Bingham, Scott (Committee member) / Nielsen, David (Committee member) / Arizona State University (Publisher)
Created2013
Description
The consumption of feedstocks from agriculture and forestry by current biofuel production has raised concerns about food security and land availability. In the meantime, intensive human activities have created a large amount of marginal lands that require management. This study investigated the viability of aligning land management with biofuel production on marginal lands. Biofuel crop production on two types of marginal lands, namely urban vacant lots and abandoned mine lands (AMLs), were assessed. The investigation of biofuel production on urban marginal land was carried out in Pittsburgh between 2008 and 2011, using the sunflower gardens developed by a Pittsburgh non-profit as an example. Results showed that the crops from urban marginal lands were safe for biofuel. The crop yield was 20% of that on agricultural land while the low input agriculture was used in crop cultivation. The energy balance analysis demonstrated that the sunflower gardens could produce a net energy return even at the current low yield. Biofuel production on AML was assessed from experiments conducted in a greenhouse for sunflower, soybean, corn, canola and camelina. The research successfully created an industrial symbiosis by using bauxite as soil amendment to enable plant growth on very acidic mine refuse. Phytoremediation and soil amendments were found to be able to effectively reduce contamination in the AML and its runoff. Results from this research supported that biofuel production on marginal lands could be a unique and feasible option for cultivating biofuel feedstocks.
ContributorsZhao, Xi (Author) / Landis, Amy (Thesis advisor) / Fox, Peter (Committee member) / Chester, Mikhail (Committee member) / Arizona State University (Publisher)
Created2013
Description
Overall, biofuels play a significant role in future energy sourcing and deserve thorough researching and examining for their best use in achieving sustainable goals. National and state policies are supporting biofuel production as a sustainable option without a holistic view of total impacts. The analysis from this research connects to policies based on life cycle sustainability to identify other environmental impacts beyond those specified in the policy as well as ethical issues that are a concern. A Life cycle assessment (LCA) of switchgrass agriculture indicates it will be challenging to meet U.S. Renewable Fuel Standards with only switchgrass cellulosic ethanol, yet may be used for California's Low Carbon Fuel Standard. Ethical dilemmas in food supply, land conservation, and water use can be connected to biofuel production and will require evaluation as policies are created. The discussions around these ethical dilemmas should be had throughout the process of biofuel production and policy making. Earth system engineering management principles can help start the discussions and allow anthropocentric and biocentric viewpoints to be heard.
ContributorsHarden, Cheyenne (Author) / Landis, Amy E. (Thesis advisor) / Allenby, Braden (Committee member) / Khanna, Vikas (Committee member) / Arizona State University (Publisher)
Created2014
Description
The production of monomer compounds for synthesizing plastics has to date been largely restricted to the petroleum-based chemical industry and sugar-based microbial fermentation, limiting its sustainability and economic feasibility. Cyanobacteria have, however, become attractive microbial factories to produce renewable fuels and chemicals directly from sunlight and CO2. To explore the feasibility of photosynthetic production of (S)- and (R)-3-hydroxybutyrate (3HB), building-block monomers for synthesizing the biodegradable plastics polyhydroxyalkanoates and precursors to fine chemicals, synthetic metabolic pathways have been constructed, characterized and optimized in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis 6803). Both types of 3HB molecules were produced and readily secreted from Synechocystis cells without over-expression of transporters. Additional inactivation of the competing PHB biosynthesis pathway further promoted the 3HB production. Analysis of the intracellular acetyl-CoA and anion concentrations in the culture media indicated that the phosphate consumption during the photoautotrophic growth and the concomitant elevated acetyl-CoA pool acted as a key driving force for 3HB biosynthesis in Synechocystis. Fine-tuning of the gene expression levels via strategies, including tuning gene copy numbers, promoter engineering and ribosome binding site optimization, proved critical to mitigating metabolic bottlenecks and thus improving the 3HB production. One of the engineered Synechocystis strains, namely R168, was able to produce (R)-3HB to a cumulative titer of ~1600 mg/L, with a peak daily productivity of ~200 mg/L, using light and CO2 as the sole energy and carbon sources, respectively. Additionally, in order to establish a high-efficiency transformation protocol in cyanobacterium Synechocystis 6803, methyltransferase-encoding genes were cloned and expressed to pre-methylate the exogenous DNA before Synechocystis transformation. Eventually, the transformation efficiency was increased by two orders of magnitude in Synechocystis. This research has demonstrated the use of cyanobacteria as cell factories to produce 3HB directly from light and CO2, and developed new synthetic biology tools for cyanobacteria.
ContributorsWang, Bo (Author) / Meldrum, Deirdre R (Thesis advisor) / Zhang, Weiwen (Committee member) / Sandrin, Todd R. (Committee member) / Nielsen, David R (Committee member) / Arizona State University (Publisher)
Created2014
Description
Second-generation biofuel feedstocks are currently grown in land-based systems that use valuable resources like water, electricity and fertilizer. This study investigates the potential of near-shore marine (ocean) seawater filtration as a source of planktonic biomass for biofuel production. Mixed marine organisms in the size range of 20µm to 500µm were isolated from the University of California, Santa Barbara (UCSB) seawater filtration system during weekly backwash events between the months of April and August, 2011. The quantity of organic material produced was determined by sample combustion and calculation of ash-free dry weights. Qualitative investigation required density gradient separation with the heavy liquid sodium metatungstate followed by direct transesterification and gas chromatography with mass spectrometry (GC-MS) of the fatty acid methyl esters (FAME) produced. A maximum of 0.083g/L of dried organic material was produced in a single backwash event and a study average of 0.036g/L was calculated. This equates to an average weekly value of 7,674.75g of dried organic material produced from the filtration of approximately 24,417,792 liters of seawater. Temporal variations were limited. Organic quantities decreased over the course of the study. Bio-fouling effects from mussel overgrowth inexplicably increased production values when compared to un-fouled seawater supply lines. FAMEs (biodiesel) averaged 0.004% of the dried organic material with 0.36ml of biodiesel produced per week, on average. C16:0 and C22:6n3 fatty acids comprised the majority of the fatty acids in the samples. Saturated fatty acids made up 30.71% to 44.09% and unsaturated forms comprised 55.90% to 66.32% of the total chemical composition. Both quantities and qualities of organics and FAMEs were unrealistic for use as biodiesel but sample size limitations, system design, geographic and temporal factors may have impacted study results.
ContributorsPierre, Christophe (Author) / Olson, Larry (Thesis advisor) / Sommerfeld, Milton (Committee member) / Brown, Albert (Committee member) / Arizona State University (Publisher)
Created2011
Description
On average, our society generates ~0.5 ton of municipal solid waste per person annually. Biomass waste can be gasified to generate synthesis gas (syngas), a gas mixture consisting predominantly of CO, CO2, and H2. Syngas, rich in carbon and electrons, can fuel the metabolism of carboxidotrophs, anaerobic microorganisms that metabolize CO (a toxic pollutant) and produce biofuels (H2, ethanol) and commodity chemicals (acetate and other fatty acids). Despite the attempts for commercialization of syngas fermentation by several companies, the metabolic processes involved in CO and syngas metabolism are not well understood. This dissertation aims to contribute to the understanding of CO and syngas fermentation by uncovering key microorganisms and understanding their metabolism. For this, microbiology and molecular biology techniques were combined with analytical chemistry analyses and deep sequencing techniques. First, environments where CO is commonly detected, including the seafloor, volcanic sand, and sewage sludge, were explored to identify potential carboxidotrophs. Since carboxidotrophs from sludge consumed CO 1000 faster than those in nature, mesophilic sludge was used as inoculum to enrich for CO- and syngas- metabolizing microbes. Two carboxidotrophs were isolated from this culture: an acetate/ethanol-producer 99% phylogenetically similar to Acetobacterium wieringae and a novel H2-producer, Pleomorphomonas carboxidotrophicus sp. nov. Comparison of CO and syngas fermentation by the CO-enriched culture and the isolates suggested mixed-culture syngas fermentation as a better alternative to ferment CO-rich gases. Advantages of mixed cultures included complete consumption of H2 and CO2 (along with CO), flexibility under different syngas compositions, functional redundancy (for acetate production) and high ethanol production after providing a continuous supply of electrons. Lastly, dilute ethanol solutions, typical of syngas fermentation processes, were upgraded to medium-chain fatty acids (MCFA), biofuel precursors, through the continuous addition of CO. In these bioreactors, methanogens were inhibited and Peptostreptococcaceae and Lachnospiraceae spp. most likely partnered with carboxidotrophs for MCFA production. These results reveal novel microorganisms capable of effectively consuming an atmospheric pollutant, shed light on the interplay between syngas components, microbial communities, and metabolites produced, and support mixed-culture syngas fermentation for the production of a wide variety of biofuels and commodity chemicals.
ContributorsEsquivel Elizondo, Sofia Victoria (Author) / Krajmalnik-Brown, Rosa (Thesis advisor) / Rittmann, Bruce E. (Committee member) / Delgado, Anca G. (Committee member) / Torres, Cesar I. (Committee member) / Arizona State University (Publisher)
Created2017
Description
To date, the production of algal biofuels is not economically sustainable due to the cost of production and the low cost of conventional fuels. As a result, interest has been shifting to high value products in the algae community to make up for the low economic potential of algal biofuels. The economic potential of high-value products does not however, eliminate the need to consider the environmental impacts. The majority of the environmental impacts associated with algal biofuels overlap with algal bioproducts in general (high-energy dewatering) due to the similarities in their production pathways. Selecting appropriate product sets is a critical step in the commercialization of algal biorefineries.
This thesis evaluates the potential of algae multiproduct biorefineries for the production of fuel and high-value products to be economically self-sufficient and still contribute to climate change mandates laid out by the government via the Energy Independence and Security Act (EISA) of 2007. This research demonstrates:
1) The environmental impacts of algal omega-3 fatty acid production can be lower than conventional omega-3 fatty acid production, depending on the dewatering strategy.
2) The production of high-value products can support biofuels with both products being sold at prices comparable to 2016 prices.
3) There is a tradeoff between revenue and fuel production
4) There is a tradeoff between the net energy ratio of the algal biorefinery and the economic viability due to the lower fuel production in a multi-product model that produces high-value products and diesel vs. the lower economic potential from a multi-product model that just produces diesel.
This work represents the first efforts to use life cycle assessment and techno-economic analysis to assess the economic and environmental sustainability of an existing pilot-scale biorefinery tasked with the production of high-value products and biofuels. This thesis also identifies improvements for multiproduct algal biorefineries that will achieve environmentally sustainable biofuel and products while maintaining economic viability.
This thesis evaluates the potential of algae multiproduct biorefineries for the production of fuel and high-value products to be economically self-sufficient and still contribute to climate change mandates laid out by the government via the Energy Independence and Security Act (EISA) of 2007. This research demonstrates:
1) The environmental impacts of algal omega-3 fatty acid production can be lower than conventional omega-3 fatty acid production, depending on the dewatering strategy.
2) The production of high-value products can support biofuels with both products being sold at prices comparable to 2016 prices.
3) There is a tradeoff between revenue and fuel production
4) There is a tradeoff between the net energy ratio of the algal biorefinery and the economic viability due to the lower fuel production in a multi-product model that produces high-value products and diesel vs. the lower economic potential from a multi-product model that just produces diesel.
This work represents the first efforts to use life cycle assessment and techno-economic analysis to assess the economic and environmental sustainability of an existing pilot-scale biorefinery tasked with the production of high-value products and biofuels. This thesis also identifies improvements for multiproduct algal biorefineries that will achieve environmentally sustainable biofuel and products while maintaining economic viability.
ContributorsBarr, William James (Author) / Landis, Amy E. (Thesis advisor) / Westerhoff, Paul (Thesis advisor) / Rittmann, Bruce (Committee member) / Khanna, Vikas (Committee member) / Arizona State University (Publisher)
Created2016
Description
Unmanned aerial vehicles have received increased attention in the last decade due to their versatility, as well as the availability of inexpensive sensors (e.g. GPS, IMU) for their navigation and control. Multirotor vehicles, specifically quadrotors, have formed a fast growing field in robotics, with the range of applications spanning from surveil- lance and reconnaissance to agriculture and large area mapping. Although in most applications single quadrotors are used, there is an increasing interest in architectures controlling multiple quadrotors executing a collaborative task. This thesis introduces a new concept of control involving more than one quadrotors, according to which two quadrotors can be physically coupled in mid-flight. This concept equips the quadro- tors with new capabilities, e.g. increased payload or pursuit and capturing of other quadrotors. A comprehensive simulation of the approach is built to simulate coupled quadrotors. The dynamics and modeling of the coupled system is presented together with a discussion regarding the coupling mechanism, impact modeling and additional considerations that have been investigated. Simulation results are presented for cases of static coupling as well as enemy quadrotor pursuit and capture, together with an analysis of control methodology and gain tuning. Practical implementations are introduced as results show the feasibility of this design.
ContributorsLarsson, Daniel (Author) / Artemiadis, Panagiotis (Thesis advisor) / Marvi, Hamidreza (Committee member) / Berman, Spring (Committee member) / Arizona State University (Publisher)
Created2016
Description
The basic scheme for photosynthesis suggests the two photosystems existing in parity with one another. However, cyanobacteria typically maintain significantly more photosystem I (PSI) than photosystem II (PSII) complexes. I set out to evaluate this disparity through development and analysis of multiple mutants of the genetically tractable cyanobacterium Synechocystis sp. PCC 6803 that exhibit a range of expression levels of the main proteins present in PSI (Chapter 2). One hypothesis was that the higher abundance of PSI in this organism is used to enable more cyclic electron flow (CEF) around PSI to contribute to greater ATP synthesis. Results of this study show that indeed CEF is enhanced by the high amount of PSI present in WT. On the other hand, mutants with less PSI and less cyclic electron flow appeared able to maintain healthy levels of ATP synthesis through other compensatory mechanisms. Reduction in PSI abundance is naturally associated with reduced chlorophyll content, and mutants with less PSI showed greater primary productivity as light intensity increased due to increased light penetration in the cultures. Another question addressed in this research project involved the effect of deletion of flavoprotein 3 (an electron sink for PSI-generated electrons) from mutant strains that produce and secrete a fatty acid (Chapter 3). Removing Flv3 increased fatty acid production, most likely due to increased abundance of reducing equivalents that are key to fatty acid biosynthesis. Additional components of my dissertation research included examination of alkane biosynthesis in Synechocystis (Chapter 4), and effects of attempting to overexpress fibrillin genes for enhancement of stored compounds (Chapter 5). Synechocystis is an excellent platform for metabolic engineering studies with its photosynthetic capability and ease of genetic alteration, and the presented research sheds light on multiple aspects of its fundamental biology.
ContributorsMoore, Vickie (Author) / Vermaas, Willem (Thesis advisor) / Wang, Xuan (Committee member) / Roberson, Robert (Committee member) / Gaxiola, Roberto (Committee member) / Bingham, Scott (Committee member) / Arizona State University (Publisher)
Created2017
Description
ABSTRACT
Sustainable global energy production is one of the grand challenges of the 21st century. Next-generation renewable energy sources include using photosynthetic microbes such as cyanobacteria for efficient production of sustainable fuels from sunlight. The cyanobacterium Synechocystis PCC 6803 (Synechocystis) is a genetically tractable model organism for plant-like photosynthesis that is used to develop microbial biofuel technologies. However, outside of photosynthetic processes, relatively little is known about the biology of microbial phototrophs such as Synechocystis, which impairs their development into market-ready technologies. My research objective was to characterize strategic aspects of Synechocystis biology related to its use in biofuel production; specifically, how the cell surface modulates the interactions between Synechocystis cells and the environment. First, I documented extensive biofouling, or unwanted biofilm formation, in a 4,000-liter roof-top photobioreactor (PBR) used to cultivate Synechocystis, and correlated this cell-binding phenotype with changes in nutrient status by developing a bench-scale assay for axenic phototrophic biofilm formation. Second, I created a library of mutants that lack cell surface structures, and used this biofilm assay to show that mutants lacking the structures pili or S-layer have a non-biofouling phenotype. Third, I analyzed the transcriptomes of cultures showing aggregation, another cell-binding phenotype, and demonstrated that the cells were undergoing stringent response, a type of conserved stress response. Finally, I used contaminant Consortia and statistical modeling to test whether Synechocystis mutants lacking cell surface structures could reduce contaminant growth in mixed cultures. In summary, I have identified genetic and environmental means of manipulating Synechocystis strains for customized adhesion phenotypes, for more economical biomass harvesting and non-biofouling methods. Additionally, I developed a modified biofilm assay and demonstrated its utility in closing a key gap in the field of microbiology related to axenic phototrophic biofilm formation assays. Also, I demonstrated that statistical modeling of contaminant Consortia predicts contaminant growth across diverse species. Collectively, these findings serve as the basis for immediately lowering the cost barrier of Synechocystis biofuels via a more economical biomass-dewatering step, and provide new research tools for improving Synechocystis strains and culture ecology management for improved biofuel production.
Sustainable global energy production is one of the grand challenges of the 21st century. Next-generation renewable energy sources include using photosynthetic microbes such as cyanobacteria for efficient production of sustainable fuels from sunlight. The cyanobacterium Synechocystis PCC 6803 (Synechocystis) is a genetically tractable model organism for plant-like photosynthesis that is used to develop microbial biofuel technologies. However, outside of photosynthetic processes, relatively little is known about the biology of microbial phototrophs such as Synechocystis, which impairs their development into market-ready technologies. My research objective was to characterize strategic aspects of Synechocystis biology related to its use in biofuel production; specifically, how the cell surface modulates the interactions between Synechocystis cells and the environment. First, I documented extensive biofouling, or unwanted biofilm formation, in a 4,000-liter roof-top photobioreactor (PBR) used to cultivate Synechocystis, and correlated this cell-binding phenotype with changes in nutrient status by developing a bench-scale assay for axenic phototrophic biofilm formation. Second, I created a library of mutants that lack cell surface structures, and used this biofilm assay to show that mutants lacking the structures pili or S-layer have a non-biofouling phenotype. Third, I analyzed the transcriptomes of cultures showing aggregation, another cell-binding phenotype, and demonstrated that the cells were undergoing stringent response, a type of conserved stress response. Finally, I used contaminant Consortia and statistical modeling to test whether Synechocystis mutants lacking cell surface structures could reduce contaminant growth in mixed cultures. In summary, I have identified genetic and environmental means of manipulating Synechocystis strains for customized adhesion phenotypes, for more economical biomass harvesting and non-biofouling methods. Additionally, I developed a modified biofilm assay and demonstrated its utility in closing a key gap in the field of microbiology related to axenic phototrophic biofilm formation assays. Also, I demonstrated that statistical modeling of contaminant Consortia predicts contaminant growth across diverse species. Collectively, these findings serve as the basis for immediately lowering the cost barrier of Synechocystis biofuels via a more economical biomass-dewatering step, and provide new research tools for improving Synechocystis strains and culture ecology management for improved biofuel production.
ContributorsAllen, Rebecca Custer (Author) / Curtiss Iii, Roy (Thesis advisor) / Krajmalnik-Brown, Rosa (Thesis advisor) / Rittmann, Bruce E. (Committee member) / Vermaas, Willem (Committee member) / Arizona State University (Publisher)
Created2016