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Simulation results for "Integration of Plant and Microbial Oil Processing at Oilcane Biorefineries for More Sustainable Biofuel Production" publication.
High-throughput amplicon sequencing of the ITS, 16S and 18S regions from soil, roots and leaves DNAs of the 50 plots present in the BCSE (Biofuel Cropping System Experiment) experiment at Kellogg Biological Station.
Meta-amplicon sequencing of the soil bacterial community of Switchgrass (var. Cave-n-Rock) along with other biofuel grasses with the Great Lakes Bioenergy Research Center Marginal Lands Experiment. Samples were collected in 2018.
The invention provides methods for engineering plants to have reduced levels of acetylation by decreasing expression of one or more Cas1L genes. Such plants can be used, e.g., to increase yield for biofuel production.
The present invention relates to terpene synthases capable of degrading precursors into biofuel compounds, such as terpenoid compounds. In one instance, a transformed organism can include such terpene synthases, as well as vectors encoding such synthases. Methods of employing such synthases and organisms are also described herein.
To achieve the economic goal of producing biofuels that can compete with petroleum-based products, we increased titers of 2,3-butanediol (BDO) production. BDO has many advantages over ethanol; including lower toxicity, higher heat of capacity, and is a precursor to various valuable applications. To produce BDO for fuel upgrading, we have genetically modified Zymomonas mobilis (YC-1). Several materials and feed strategies were evaluated to determine the best titer and economic feasibility. Titers from DMR corn stover in a batch process were in the range of 70-80 g/L. The fully hydrolyzed material was easy to work with but suffered from dilution issues. We pushed the concentration of solids to above 25% w/w and were met with numerous challenges. This included an initial sugar concentration above 150 g/L, the high solid concentration altered the oxygen mass transfer rate and affected which products were produced, and the fermentation conditions were not optimal for the enzyme. Despite these challenges, our current results yielded 100 g/L of upgradeable products (70 g/L BDO and 30 g/L acetoin) and 83% fermentation process yield. However, the poor enzymatic hydrolysis yield of 64%, unconverted xylose, and the high acetoin levels warrant further exploration for optimization. Using unhydrolyzed solids as the feed material, we achieved titers in the range of 75-95 g/L BDO. The raw solids feed, though operationally more challenging to work with, proved to be promising as the rate of hydrolysis promoted co-utilization of the sugars. We reached 150 g/L BDO using concentrated DDR liquors under boules and continuous feeding strategies. The clarified liquors showed high titers, high productivity, high yield, and high sugar utilization with little drawback operationally. The target goal of under $3.50 GGE was achieved in several feeding strategies. Scale-up feasibility has also been demonstrated at the 100L scale using both whole slurry and liquor hydrolysates.
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Utilizing biomass resources, such as cellulosic biomass and waste, can greatly contribute to decarbonization efforts in the U.S. economy. The U.S. bioenergy sector includes corn ethanol production, biodiesel, renewable diesel production, and the utilization of biomass wastes for electricity generation being the primary applications. Within the electricity sector, biopower can play a crucial role as a stable low-carbon resource. Enhancing the electricity mix's diversity could enhance grid reliability. If the issues regarding hot gas cleanup can be resolved, flexible biopower resources like biomass gasification facilities could complement the integration of variable renewable energy sources due to their quick ramp-up and ramp-down times. The criticality of bioenergy deployment lies in its ability to decarbonize hard-to-electrify sectors, such as aviation, where alternative decarbonization options may not be viable in the short term. Moreover, bioenergy has the potential to be converted into process heating, building materials, and plastics, which are not considered in this study. A set of pathways was carefully chosen to represent viable options for converting ample herbaceous and woody cellulosic feedstocks into fuels, chemicals, and electricity in this study. In conclusion, biomass pathways provide flexibility by generating various types of bioenergy and bioproducts, including electricity, hydrogen, liquid fuels, biochemicals, and bioplastics. When paired with carbon dioxide capture and storage (CCS), specific bioenergy approaches can effectively extract carbon dioxide from the atmosphere, thus providing an effective decarbonization option for the transportation sector.
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Thermophilic microbes are an attractive bioproduction platform due to their inherently lower contamination risk and their ability to perform thermostable enzymatic processes which may be required for biomass processing and other industrial applications. The engineering of microbes for industrial scale processes requires a suite of genetic engineering tools to optimize existing biological systems as well as to design and incorporate new metabolic pathways within strains. Yet, such tools are often lacking and/or inadequate for novel microbes, especially thermophiles. This chapter focuses on genetic tool development and engineering strategies, in addition to challenges, for thermophilic microbes. We provide detailed instructions and techniques for tool development for an anaerobic thermophile, Caldanaerobacter subterraneus subsp. tengcongensis, including culturing, plasmid construction, transformation, and selection. This establishes a foundation for advanced genetic tool development necessary for the metabolic engineering of this microbe and potentially other thermophilic organisms.
This study evaluates cover crops in Washington State (WA) as a renewable feedstock for hydrothermal liquefaction (HTL), supported by techno-economic (TEA) and life cycle (LCA) analysis. In the Pacific Northwest, fallow land is the common winter practice, generating no income for farmers. Over three years, field trials at two representative sites—Puyallup (cool, wet) and Othello (irrigated, no-till)tested triticale, hairy vetch, crimson clover, winter pea, and fava bean preceding cash crops. Biomass yields, composition, soil impacts, and net revenue were assessed across removal treatments. HTL conversion produced 32–37 wt% biocrude, lower than 48 wt% for sewage sludge, with modeled cradle-to-grave carbon intensities of 26–31 g CO 2 e/MJ and minimum fuel selling prices (MFSP) of $\$$6.75–$\$$7.18 per gallon gasoline equivalent (GGE), comparable to other lignocellulosic feedstocks. No significant changes were observed in soil carbon, nitrogen, or subsequent cash crop yields after cover crop removal. Farmer profitability was possible, particularly for triticale and hairy vetch at $\$$60 per dry ton. To address seasonal variability, blending with wastewater sludge was evaluated. All scenarios delivered >70 % carbon intensity reductions relative to fossil fuels, and with Renewable Identification Number (RIN) credits, MFSPs below $\$$2.5 GGE were achievable. These results demonstrate that cover crops can generate new revenue streams for farmers while serving as a flexible, low-carbon feedstock. Integrating cover crops with waste biomass offers a practical pathway to expand the bioeconomy and support year-round renewable fuel production.
Advanced catalytic materials play an enabling role in producing renewable fuels and chemicals from biomass, thereby helping meet the global climate-change goals set forth by the Intergovernmental Panel on Climate Change. Herein, we present a multiscale approach to accelerate the catalyst-process development cycle for catalytic fast pyrolysis (CFP) of biomass over Mo2C. Mo2C has been shown to possess co-localized acidic and metallic sites and exhibit high activity for deoxygenation of biomass pyrolysis model compounds. However, critical knowledge gaps remain regarding the effectiveness of this catalyst for CFP of whole biomass. We address these knowledge gaps and demonstrate that Mo2C is effective at deoxygenating biomass-pyrolysis products in the presence of H2 but that it undergoes rapid selective and non-selective deactivation. The knowledge gaps addressed from this integrated study, targeting appropriate experiments across scales and feed types, enabled identification of critical modifications for advancing the CFP catalyst-process development cycle.
Hydrothermal liquefaction (HTL) is the process in which biological compounds can be converted into biocrude diesel; various feedstocks, such as algae and wet biowaste, can be utilized. Almost 20 million dry tons of wet biowaste is deposited in U.S. municipal wastewaters annually. If recovered and converted into biodiesel, it can act as a massive source of renewable energy. Preliminary experiments on a diesel engine show that HTL fuel can achieve similar performance and emission levels to diesel. In order to further understand the influence of the HTL fuel properties, the HTL fuel and blends with diesel are observed in a constant volume chamber. Previously, 10% and 20% of the HTL fuel were blended with diesel by volume and demonstrated properties that roughly matched diesel in terms of combustion. In this study, 50% and 100% HTL fuel blends are tested on the same apparatus. It was discovered that the workability of HTL is almost the same as that of diesel and is less sensitive to ambient temperatures. When the ambient temperature is reduced from 1200 K to 800 K, the change ratio of ignition delay is only 62% of diesel. By analyzing the combustion process, it can be seen that there is no need to make major modifications to the engine control strategy if HTL fuel is applied to diesel engines. Though, under low temperature and low oxygen conditions, the soot luminosity peak of HTL100 is 3.12 times than that of diesel.
Here, we present the results of a lab-scaled feasibility study to assess the performance of electrical resistivity tomography for detection, characterization, and monitoring of fuel grade ethanol releases to the subsurface. Further, we attempt to determine the concentration distribution of the ethanol from the electrical resistivity tomography data using mixing-models. Ethanol is a renewable fuel source as well as an oxygenate fuel additive currently used to replace the known carcinogen methyl tert-butyl ether; however, ethanol is preferentially biodegraded and a cosolvent. When introduced to areas previously impacted by nonethanol-based fuels, it will facilitate the persistence of carcinogenic fuel compounds like benzene and ethylbenzene, as well as remobilize them to the ground water. These compounds would otherwise be retained in the soil column undergoing active or passive remediation processes such as soil vapor extraction or natural attenuation. Here, we introduce ethanol to a saturated Ottawa sand in a tank instrumented for four-dimensional geoelectrical measurements. Forward model results suggest pure phase ethanol released into a water saturated silica sand should present a detectable target for electrical resistivity tomography relative to a saturated silica sand only. We observe the introduction of ethanol to the closed hydraulic system and subsequent migration over the duration of the experiment. One-dimensional and three–dimensional temporal data are assessed for the detection, characterization, and monitoring of the ethanol release. Results suggest one-dimensional geoelectrical measurements may be useful for monitoring a release, while three-dimensional geoelectrical field imaging would be useful to characterize, monitor, and design effective remediation approaches for an ethanol release, assuming field conditions do not preclude the application of geoelectrical methods. We then attempt to use predictive mixing models to calculate the distribution of ethanol concentration within the measurement domain. For this study we examine four different models: a nested parallel mixing model, a nested cubic mixing model, the complex refractive index model (CRIM), and the Lichtenecker-Rother (L-R) model. The L-R model, modified to include an electrical formation factor geometry term, provided the best agreement with expected EtOH concentrations.
Cyclopropane-functionalized hydrocarbons are excellent fuels due their high energy density. However, the organic synthesis of these molecules is challenging and harmful to the environment. In this work we produced polycyclopropanated fatty acids in bacteria. These molecules can be easily converted into renewable fuels for high energy applications such as shipping, long-haul transport, aviation, and rocketry. We explored the chemical diversity encoded in the genome of thousands of bacteria to identify and repurpose naturally occurring cyclopropanated molecules. We identified a set of candidate iterative polyketide synthases (iPKSs) predicted to produce polycyclopropanated fatty acids (POP-FAs), expressed these PKSs in Streptomyces coelicolor and produced the POP-FAs. We determined the structure of the molecules and increased their production 22-fold. Polycyclopropanated fatty acid methyl esters (POP-FAMEs) were obtained by methyl esterifying the POP-FAs. Finally, we calculated that our POP fuel candidates can have energy densities of more than 50 MJ/L. Our research shows that POP-FAMEs and other polyketide derived POPs have the energetic properties for energy-demanding applications for which sustainable alternatives are scarce.