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At least 109 records · Page 6

Varied farm-level carbon intensities of corn feedstock help reduce corn ethanol greenhouse gas emissions

Abstract A reduction in the overall carbon intensity (CI) of a crop-based biofuel can be achieved by cutting down the CI of the biofuel’s feedstock, which in turn correlates significantly to agricultural management practices. Proposals are being made to incentivize low-carbon biofuel feedstocks under U.S. fuel regulatory programs to promote sustainable farming practices by individual farms. For such an incentive scheme to function properly, robust data collection and verification are needed at the farm level. This study presents our collaboration with U.S. private sector companies to collect and verify the corn production data necessary for feedstock-specific CI calculation at the farm level, through a carefully designed questionnaire, to demonstrate the practicality and feasibility of data collection at scale. We surveyed 71 farms that produced 0.2 million metric tons of corn grain in 2018 in a Midwestern U.S. state to obtain information on key parameters affecting corn ethanol feedstock CI, such as grain yields, fertilizer/chemical application rates, and agronomic practices. Feedstock-specific CI was calculated in the unit of grams (g) CO 2 equivalent (CO 2 e) of greenhouse gases per kilogram (kg) of corn produced. Results showed large CI variations—from 119 to 407 g CO 2 e kg −1 of corn—due to the farm-level inventory, while the production-weighted average CI for all surveyed farms was 210 g CO 2 e kg −1 , comparable to the national average CI of 204 g CO 2 e kg −1 . The nitrogen fertilizer type applied and rate were identified as key factors contributing most to CI variations at the farm level. The estimated N 2 O emissions from fertilizer and biomass nitrogen inputs to soil accounted for 51% of the overall farm-level CI and therefore need to be better monitored at farm level with high resolution. We concluded that this feedstock-specific, farm-level CI evaluation has the potential to be used to incentivize low-carbon feedstock for biofuel production.

54 ENVIRONMENTAL SCIENCES↗

Denoising diffusion probabilistic models for generative alloy design

Inverse material design is an extremely challenging optimization task made difficult by, in part, the highly nonlinear relationship linking performance with composition. Quantitative approaches have improved significantly owing to advances in high throughput experimentation and computational thermodynamics. However, existing physics-based tools are mostly forward models; input a chemistry and obtain a prediction. More recently the materials community has leveraged advances in the machine learning community to establish novel inverse design frameworks. Very recently denoising diffusion probabilistic models have been shown to be extremely powerful generators producing synthetic data of various modalities e.g. images, text, audio, tables, etc.. In this work a novel framework for alloy design and optimization is proposed leveraging these class of models. Five key generative tasks are demonstrated (1) unconditional generation (2) composition conditioned generation (3) property conditioned generation (4) multi-feedstock conditioned generation and (5) generative optimization. These methods were tested on three case studies: high entropy alloy design, superalloy binder jet additive manufacturing, and in-situ dual-feedstock wire-arc additive manufacturing. Results indicate that the established models are extremely flexible, expressive, and robust. The architecture’s flexibility and training procedure empower the model to learn complex intra-compositional and composition-property relationships. Furthermore, the probabilistic nature of these models makes them well suited for addressing solution non-uniqueness and tackling uncertainty quantification tasks. While the fidelity and quantity of the underlying training data is paramount, we envision that future alloy design frameworks will make extensive use of these kinds of machine learning models as “search” tools bolstering the utility of experimental and computational approaches.

36 MATERIALS SCIENCE↗

Performance Testing of Moving Bed Gasifier Using Biomass and Waste Fuels to Generate Low-Cost Clean Hydrogen

Our need for hydrogen is growing as the world transitions toward a low-carbon future. Hydrogen provides long-term energy storage for grid stability in a solar- and wind-dominated power market and can be used to decarbonize other sectors. One promising process for generating low-cost hydrogen that produces net-negative carbon is to gasify biomass with a mixture of legacy coal wastes, waste plastics, and other wastes with carbon capture. Use of waste fuels lowers costs and diverts waste from landfills. EPRI is leading a project, funded by the U.S. Department of Energy, to conduct performance testing of modular, moving-bed gasification for the generation of low-cost, clean hydrogen from biomass mixed with legacy coal waste, waste plastic, and/or refuse derived fuels. The work scope includes preparation of multiple pellet feedstocks using biomass (both woody biomass and corn stover) with a mixture of legacy coal waste, plastic waste (wire insulation), and refuse-derived fuel (RDF). These pelletized feedstocks are being qualified based on performance testing of selected fuel blend compositions in updraft moving-bed gasifier located in Sardinia, Italy. Testing is being conducted to obtain relevant data to advance the modular design of the moving-bed gasification process, and successfully use these feedstocks to produce a high hydrogen content raw syngas that can be shifted to produce clean hydrogen. Testing results will be used to determine the effects of the various fuels on feedstock development, the resulting products (i.e., syngas compositions, organic condensate production, and ash characteristics), and impacts on gasifier operations.

08 HYDROGEN↗

Major element composition controls rare earth element solubility during leaching of coal fly ash and coal by-products

Coal combustion ash and pre-combustion coal refuse are currently under consideration as potential sources of rare earth elements. One of the early steps in recovering REEs from coal by-products is often acid leaching, which can result in low pH leachates with complex aqueous chemistry. The aim of this work was to understand the connection between REE solubility, pH, and major elemental components of leachates for coal by-products. To accomplish this, we investigated the effects of solids concentration (i.e., pulp density) and pH adjustment on REE solubility in acid leachates of coal fly ashes from the Powder River Basin (PRB) and Appalachian Basin in the United States, and a coal processing refuse from the Southwestern U.S. For PRB ashes, the concentrations of soluble REEs generally increased with increasing pulp density; however, at pulp density values above 80–100 g/L, the soluble REE concentrations in the leachates were markedly lower. Similarly, the soluble concentrations of other major solutes (Fe, Al, Si) that leached from PRB fly ashes were also non-linear with pulp density. These major elements tended to reach maximum concentration values at 60–70 g/L pulp density. In contrast, for the Appalachian fly ashes and the coal by-product, soluble concentrations of REE and major elements in leachates increased linearly with pulp density. Chemical equilibria calculations of mineral saturation indices indicated that trends in soluble REE concentrations could be explained by saturation conditions for Fe and Al-(hydr)oxides and possibly sulfate minerals, but not lanthanide hydroxides. Furthermore, pH adjustment of the acid leachates showed that REEs and many major solutes were removed from solution at pH values above 4.5, also consistent with Fe- and Al-(hydr)oxide precipitation. These results highlight the importance of understanding the chemical composition of leachates when designing REE recovery processes for low-grade geologic feedstocks and that precipitation of hydr(oxide) or sulfate minerals of major elements rather than discreet formation of REE mineral phases could be used for process optimization.

01 COAL, LIGNITE, AND PEAT↗

Metabolic engineering to improve production of 3-hydroxypropionic acid from corn-stover hydrolysate in Aspergillus species

Fuels and chemicals derived from non-fossil sources are needed to lessen human impacts on the environment while providing a healthy and growing economy. 3-hydroxypropionic acid (3-HP) is an important chemical building block that can be used for many products. Biosynthesis of 3-HP is possible; however, low production is typically observed in those natural systems. Biosynthetic pathways have been designed to produce 3-HP from a variety of feedstocks in different microorganisms. In this study, the 3-HP β-alanine pathway consisting of aspartate decarboxylase, β-alanine-pyruvate aminotransferase, and 3-hydroxypropionate dehydrogenase from selected microorganisms were codon optimized for Aspergillus species and placed under the control of constitutive promoters. The pathway was introduced into Aspergillus pseudoterreus and subsequently into Aspergillus niger, and 3-HP production was assessed in both hosts. A. niger produced higher initial 3-HP yields and fewer co-product contaminants and was selected as a suitable host for further engineering. Proteomic and metabolomic analysis of both Aspergillus species during 3-HP production identified genetic targets for improvement of flux toward 3-HP including pyruvate carboxylase, aspartate aminotransferase, malonate semialdehyde dehydrogenase, succinate semialdehyde dehydrogenase, oxaloacetate hydrolase, and a 3-HP transporter. Overexpression of pyruvate carboxylase improved yield in shake-flasks from 0.09 to 0.12 C-mol 3-HP C-mol -1 glucose in the base strain expressing 12 copies of the β-alanine pathway. Deletion or overexpression of individual target genes in the pyruvate carboxylase overexpression strain improved yield to 0.22 C-mol 3-HP C-mol -1 glucose after deletion of the major malonate semialdehyde dehydrogenase. Further incorporation of additional β-alanine pathway genes and optimization of culture conditions (sugars, temperature, nitrogen, phosphate, trace elements) for 3-HP production from deacetylated and mechanically refined corn stover hydrolysate improved yield to 0.48 C-mol 3-HP C-mol -1 sugars and resulted in a final titer of 36.0 g/L 3-HP. The results of this study establish A. niger as a host for 3-HP production from a lignocellulosic feedstock in acidic conditions and demonstrates that 3-HP titer and yield can be improved by a broad metabolic engineering strategy involving identification and modification of genes participated in the synthesis of 3-HP and its precursors, degradation of intermediates, and transport of 3-HP across the plasma membrane.

09 BIOMASS FUELS↗

Designing Harvesting and Hauling Cost Models for Energy Cane Production for Biorefineries

The harvesting and hauling operations of bioenergy feedstock is an important area in biofuel production. Production costs can be minimized by maintaining optimal machinery units for these operations. The objective of this study is to design an optimal harvesting unit for bioenergy refinery and estimate harvesting and hauling costs of energy cane. A biorefinery with the annual capacity of processing twenty-five million imp. gallons of ethanol were considered. Given the efficiency of harvesting, a two-row soldier system was considered. Considering the year-round supply of energy cane to the refinery, the optimal machinery unit was designed, and the combined operation costs were derived. The average estimated ownership, repair, labor and fuel and lubricant costs of biomass harvest unit were calculated to be $\$$0.50, $\$$0.54, $\$$1.78 and $\$$1.51/mt, respectively. The costs distribution generated showed harvesting and hauling costs could range between $\$$5.47–$\$$9.23/mt of energy cane. The methodology and the research output will provide guidelines for investors in designing harvesting and hauling units and estimating costs for different scales of operation.

09 BIOMASS FUELS↗

Trace Element Control in Master Alloys and Impact of Feedstock Purity on a Ta-containing Steel during Electroslag Remelting

Advanced materials design often leads to new complex compositions that can bring challenges to manufacturing processes. Melt processing of a novel alloy with a tight chemistry range revealed a 25% Ta loss after manufacturing. Vacuum induction melting (VIM) and electroslag remelting (ESR) were performed using industry practices at a laboratory scale and the loss occurred from the VIM electrode to the ESR ingot. Several tools were used for characterization, including combustion analysis, scanning electron microscopy, and electron probe microanalysis for observation of the precipitate and inclusion phases. Additionally, computational modeling of the ESR process was performed to predict macrosegregation and inclusion travel. It was found that a significant amount of Ta2O5 inclusions formed during VIM and were transferred to the slag during ESR. This led to a 95% decrease in density of inclusions explaining to Ta loss. Pathways to better control advanced alloy chemistries during melt processing will be discussed.

Detrois, Martin↗

Performance Testing of a Moving-Bed Gasifier Using Coal, Biomass, and Waste Plastic Blends with Washed and Unwashed Legacy Coals and Other Waste Fuels to Generate White Hydrogen

The objective of this effort, primarily funded by the United States Department of Energy (DOE), and led by the Electric Power Research Institute, Inc. (EPRI), with support by Hamilton Maurer International (HMI) and Sotacarbo S.p.A. (Sotacarbo), has been to qualify coal, biomass, and plastic waste blends based on performance testing of selected fuel pellet compositions in a pilot-scale updraft moving-bed (UDMB) gasifier. The testing provided relevant data to advance the commercial-scale design of the moving-bed gasifier to be able to successfully use these feedstocks to produce hydrogen. In particular, the effects of waste plastics on feedstock development (i.e., blending and pelletizing) and the resulting products (i.e., syngas compositions, organic condensate production, and ash characteristics) are the focus. The gasifier used for testing is HMI’s moving-bed gasifier, which has been proven capable of gasifying nearly all coal ranks. It has also shown the ability in prior testing work to gasify wood chips (biomass). However, mixtures of these fuels with plastic wastes have not been prepared and gasified together. The three feedstocks were densified and pelletized by California Pellet Mill (CPM) to meet the feedstock size required by Sotacarbo’s 30mm ID UDMB gasifier, under contract to HMI. The technical tasks and results from this two-year research project included: (1) Feed Procurement and Preparation: Nine different tri-fuel pellets were prepared from varying compositions of fresh mined PRB coal, corn stover biomass, and car fluff waste plastics. Tri-fuel pellets were produced by CPM and shipped to Sotacarbo’s test facility in Carbonia, Sardinia, Italy. (2) Test Plan Development: A test plan was created to define the test runs to be performed. The test plan detailed the different UDMB gasification tests to be performed in Sotacarbo’s 12-inch ID pilot scale gasifier, the process monitoring instrumentation used, and the extractive samples recovered for analysis of the total gasification process mass and energy balance. (3) Gasifier Testing: Nine different gasification runs were performed in the pilot-scale gasifier at Sotacarbo using nine different fuel feedstock compositions generated from varying mixtures of PRB coal, biomass, and plastic wastes. The testing generated performance data on gasification reaction efficiency and performance, yielding relevant data for models used to scale up the gasifier design. This task also included work to refurbish and reassemble the pilot gasifier at Sotacarbo and perform a baseline 100% PRB coal run. (4) Data Analysis and Reporting: Review of the data, determination of figures of merit, and interpretation of the results are reported in the project’s final report, published in March 2024. The results show that all tri-fuel pellets gasified well and maintained structural integrity throughout the gasification process. The syngas generated can be shifted to hydrogen by using commercial syngas shifting technologies. (5) High Fidelity computational fluid dynamics (CFD) Simulation: The National Energy Technology Laboratory (NETL) team performed CFD simulations of the UDMB gasifier for two of the tri-fuel pellets gasified in Sotacarbo’s pilot scale gasifier. The kinetic mechanisms for the pyrolysis of each constituent, PRB coal, corn stover biomass, and waste plastics are based on thermogravimetric analysis performed by Sotacarbo. The gasification model was validated by comparing the predicted syngas composition at the exit of the gasifier with the measured syngas composition. In addition, the reactor’s measured internal temperature profile agreed well with the predicted internal reactor temperature profile. These results validate that the model can be used to predict the performance of the updraft moving bed gasifier for different feedstocks and operating conditions. This paper summarizes the results of the completed work in which the pelletizing procedure was validated to ensure the viability of the tri-fuel pellets for the gasification runs performed at Sotacarbo’s 30 mm UDMB gasifier. The gasification performance data from this series of nine runs will enable modeling of a full-scale HMI industrial scale gasifier supporting both combined heat and power, and Hydrogen production from coal (both fresh mined and legacy) combined with various biomass and waste plastics. Additionally, plans and progress on a follow-up project, being executed by the same project team, will be presented. In this project, a total of twenty (20) different feedstocks are being prepared from varying compositions of biomass (both woody biomass and corn stover) with a mixture of legacy coal waste, plastic waste, and refuse-derived fuel (RDF). The testing will provide information on gasification reaction efficiency/performance, yielding relevant data for models used to scale up the gasifier design to 50 megawatt electric (MWe) (equivalent hydrogen production). Tests will also be performed on a bench-scale fluidized-bed gasifier for comparison purposes. The results of this testing will be used to specify the range of feedstock blends that can be successfully gasified as well as quantify gasifier outputs based on specific blends.

08 HYDROGEN↗

Performance Testing of a Moving-Bed Gasifier Using Coal, Biomass, and Waste Plastic Blends to Generate White Hydrogen

The objective of this DOE-funded project by the Electric Power Research Institute, Inc. (EPRI), Hamilton Maurer International (HMI) and Sotacarbo S.p.A. (Sotacarbo), is to qualify coal, biomass, and plastic waste blends based on performance testing of selected pellet recipes in a pilot-scale updraft moving-bed gasifier. The testing will provide relevant data to advance the commercial-scale design of the moving-bed gasifier to be able to successfully use these feedstocks to produce hydrogen. In particular, the effects of the waste plastics on feedstock development (i.e., blending and pelletizing) and the resulting products (i.e., syngas compositions, organic condensate production, and ash characteristics) are a focus. The gasifier is Hamilton Mauer International, Inc. (HMI)’s moving-bed gasifier, which has been proven capable of gasifying nearly all coal ranks. It has also shown the ability in prior testing work to gasify wood chips. However, mixtures of these fuels with plastic wastes have not been prepared and gasified together. The feedstocks will be prepared by California Pellet Mill (CPM) under contract to HMI. The technical tasks and current status for this two-year research project are: Feed Procurement and Preparation: Nine different feedstocks were prepared from varying compositions of PRB coal, corn stover biomass, and car fluff waste plastics. Fuel pellets were produced by California Pellet Mill and shipped to Sotacarbo’s test facility in Italy. Test Plan Development: A test plan was created to define the test runs to be performed. The test plan detailed the different tests that were run, instrumentation used, extractive samples taken, and relevant figures of merit. Gasifier Testing: Tests are currently being performed in the pilot-scale gasifier at Sotacarbo using nine different fuel feedstocks generated from varying mixtures of coal, biomass, and plastic wastes. The testing will provide information on gasification reaction efficiency/performance, yielding relevant data for models used to scale up the gasifier design. This task will also include work to reassemble the gasifier at Sotacarbo and perform a baseline 100% coal run. Data Analysis and Reporting: Review of the data, determination of figures of merit, and interpretation of the results to be reported. The results will be used to specify the range of feedstock blends that can be successfully gasified as well as quantify gasifier outputs based on specific blends. This paper will be summarize the pelletizing procedure that insures the viability of the tri-fuel pellets for the gasification runs that are being performed at Sotacarbo’s 30mm up draft moving bed gasifier. Initial gasification tests have been conducted, and all the lab scale tri-fuel pellet gasification runs will be completed by the fall of 2022. Performance data will enable modeling of a full-scale HMI industrial scale gasifier supporting both CHP and Hydrogen production.

01 COAL, LIGNITE, AND PEAT↗

Additive manufacturing of power poles using bamboo-polymer composites: A quick solution for power grid restoration

Polymer additive manufacturing (AM) technology is rapidly growing and its transition to become an advanced manufacturing technique now offers possibilities for unconventional applications. One of these potential applications is on site additive manufacturing of power poles. During natural incidents such as storms and hurricanes, power lines can be severely damaged and there may not be sufficient spare power poles to replace. In such situations, additive manufacturing of power poles on site can offer a fast, temporary solution during emergencies. The ability of freeform manufacturing of parts with complex geometries makes large scale AM attractive for various applications; however, due to residual stress build up and need for different mechanical requirements for different applications, reinforcement of the AM feedstock is necessary. In addition to carbon and glass fibers, bio-derived fibers can also be used as a reinforcing phase, both to utilize the natural resources at the emergency area and in terms of long-term sustainability. In this project, design and additive manufacturing of a 13.1 m power pole was demonstrated. Three different feedstock material options (polystyrene, cellulose ester and polycarbonate reinforced with glass and bamboo fibers) and two different pole designs were investigated. As-printed part tensile properties both in printing and interlayer directions were measured and a power pole was printed as six pieces and put together successfully. Finally, a conceptual design of a portable power pole manufacturing approach was proposed.

Tekinalp, Halil↗

Pilot-Scale Continuous Plug-Flow Hydrothermal Liquefaction of Food Waste for Biocrude Production

Pilot-scale hydrothermal liquefaction (HTL) of biowaste is a critical step toward commercialization of the HTL technology. Despite many HTL studies conducted with wet biomass, including food waste, few were performed with a pilot-scale continuous plug-flow reactor (PFR), with the biocrude yield and quality analysis based on dewatering (ASTM D2892 Annex X1). This paper describes the development and performance evaluation of a mobile pilot-scale HTL continuous PFR, with a processing capacity of 60 L/h of wet feedstock and 6 L/h of biocrude production. The reactor system was designed for reaction conditions of up to 325 °C and 17.25 MPa. The reactor has a volume of 28.88 L with an additional counterflow heat exchanger volume of 18.07 L. Two types of food wastes, from a food processing plant and grocery store, were processed at 280 °C for 30 min, producing biocrude oil yields of 52.19 and 47.06 wt %, energy recoveries of 68.17 and 70.77%, and carbon recoveries of 66.91 and 64.78%, respectively. Furthermore, due to its high feedstock capacity and reaction volume, large amounts of biocrude oil and post-HTL wastewater (PHW) were obtained from this pilot-scale reactor to allow downstream research on upgrading biocrude oil for transportation fuel as well as PHW treatment and nutrient recovery.

09 BIOMASS FUELS↗

Biosystem design of Corynebacterium glutamicum for bioproduction

Corynebacterium glutamicum, a natural glutamate-producing bacterium adopted for industrial production of amino acids, has been extensively explored recently for high-level biosynthesis of amino acid derivatives, bulk chemicals such as organic acids and short-chain alcohols, aromatics, and natural products, including polyphenols and terpenoids. Here, we review the recent advances with a focus on biosystem design principles, metabolic characterization and modeling, omics analysis, utilization of nonmodel feedstock, emerging CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) tools for Corynebacterium strain engineering, biosensors, and novel strains of C. glutamicum. Further, future research directions for developing C. glutamicum cell factories are also discussed.

59 BASIC BIOLOGICAL SCIENCES↗

Chapter 14: Sustainable Biomass Conversion Process Assessment

Process intensification and integration for sustainable design play an essential role in addressing and alleviating severe sustainable development challenges. It is prudent to evaluate a process design in its early stage development to ensure the process can achieve sustainability goals. The primary goal of a sustainability evaluation is to assess the impact of systems on areas sought to be protected and maintained over time, such as human well-being and ecosystems (Schaubroeck, T. and Rugani, B. (2017). A revision of what life cycle sustainability assessment should entail: towards modeling the net impact on human well-being. Journal of Industrial Ecology 21 (6): 1464-1477). Parenthetically, sustainable process design to convert biomass to fuels and chemicals plays an integral part in the bioeconomy sustainability. It is recognized that the integration of sustainability in process design is core to the mission in developing renewable fuels and should be considered a necessary practice in biorefinery design. This study demonstrates the use of Gauging Reaction Effectiveness for ENvironmental Sustainability of Chemistries with a multi-Objective Process Evaluator (GREENSCOPE) as a tool to help track progress on process sustainability performance attributed to conversion improvement. With the incorporation of a wide range of indicators, GREENSCOPE captures the multi-dimensional aspect of the process design and operation. As opposed to conventional techno-economic analysis and life cycle assessment (in which the focuses are limited to economic and environmental aspects), GREENSCOPE enables the consideration of process intensification and integration for sustainable design. A high-octane gasoline production from woody biomass feedstock is used as a case study to demonstrate the approach.

biomass↗

New strategy for liquefying corn stover pellets

The movement of solid material into and between unit operations within a biorefinery is a bottleneck in reaching design capacity, with formation of biomass slurries needed to introduce feedstock. Corn stover slurries have been achieved from dilute acid, pretreated materials resulting in slurry concentrations of up to about 150 g/L, above which flowability is compromised. We report a new strategy to liquefy corn stover at higher solids concentration (300 g/L) by initially cooking it with the enzyme mimetic maleic acid at 40 mM and 150°C. This is followed by 6 h of enzymatic modification at 1 FPU (2.2 mg protein)/g solids, resulting in a yield stress of 171 Pa after 6 h and 58 Pa in 48 h compared to 6806 Pa for untreated stover. Mimetic treatment of corn stover pellets minimizes the inhibitory effect of xylo-oligomers on hydrolytic enzymes. Furthermore, this strategy allows for the delivery of solid lignocellulosic slurry into a pretreatment reactor by pumping, improving operability of a biorefinery.

09 BIOMASS FUELS↗

Technical Information Needs and Regulatory Considerations for Front-End Transportation Activities of HALEU Fuel Feed Material

This report fulfills task order number 31310022F0033 Task 1 Assessment of technical information needs and considerations for front-end transportation activities for HALEU fuel feed material. This assessment primarily focuses on the transportation outlook and packaging requirements for transporting high-assay low-enriched uranium (HALEU) fuel feed material. The regulatory framework is described in section 3.0. In section 4.0 the specific issues associated with reviewing HALEU feed material packages are discussed. The rest of the report discusses the need for new packages and Department of Energy progress on HALEU production. Through this evaluation it is shown that the 5% enrichment barrier between low-enriched uranium and HALEU is a significant challenge to transportation packaging. However, there are licensed designs and concepts that can be used to transport HALEU feedstock. No significant issues were identified through the HALEU range up to 20% enriched.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Rapid and robust squashed spore/colony PCR of industrially important fungi

Fungi have been utilized for centuries in medical, agricultural, and industrial applications. Development of systems biology techniques has enabled the design and metabolic engineering of these fungi to produce novel fuels, chemicals, and enzymes from renewable feedstocks. Many genetic tools have been developed for manipulating the genome and creating mutants rapidly. However, screening and confirmation of transformants remain an inefficient step within the design, build, test, and learn cycle in many industrial fungi because extracting fungal genomic DNA is laborious, time-consuming, and involves toxic chemicals. In this study we developed a rapid and robust technique called “Squash-PCR” to break open the spores and release fungal genomic DNA as a template for PCR. The efficacy of Squash-PCR was investigated in eleven different filamentous fungal strains. Clean PCR products with high yields were achieved in all tested fungi. Spore age and type of DNA polymerase did not affect the efficiency of Squash-PCR. However, spore concentration was found to be the crucial factor for Squash-PCR in Aspergillus niger, with the dilution of starting material often resulting in higher PCR product yield. We then further evaluated the applicability of the squashing procedure for nine different yeast strains. We found that Squash-PCR can be used to improve the quality and yield of colony PCR in comparison to direct colony PCR in the tested yeast strains. The developed technique will enhance the efficiency of screening transformants and accelerate genetic engineering in filamentous fungi and yeast.

59 BASIC BIOLOGICAL SCIENCES↗

Enabling Entrained-Flow Gasification of Blends of Coal, Biomass and Plastics (Final Report)

This project demonstrated the technical feasibility of gasifying blends of coal, biomass and mixed waste plastics in a high pressure entrained-flow gasifier for production of hydrogen. This was achieved by creating slurries of coal, biomass pyrolysis liquids and liquefied plastic oil. Specific objectives of the project included (1) determining compositions of coal-biomass-plastic mixtures that produce a stable slurry suitable for pumping to high pressure, (2) designing and testing a novel burner to effectively atomize the mixed feedstock slurry in a pressurized gasifier, and (3) acquiring first-of-a-kind performance data for pressurized, oxygen-blown entrained-flow gasification of slurried blends of coal, biomass and plastic. A key focus of the project was preparation of mixtures of coal, biomass and plastic. To facilitate easy pumping to high pressure, slurries were created using (i) bioliquid resulting from a well-established rapid thermal treatment pyrolysis process, (ii) oil produced from waste plastic via an available thermal conversion process, and (iii) ground coal. Lab-scale tests were conducted to evaluate viscosity and stability/propensity for the mixtures to separate over time. Larger batches of the most promising slurry compositions were prepared and subjected to pumping studies. It was found that mixtures containing less than about 40% coal and less than about 30% plastic oil were stable and of low enough viscosity to facilitate pumping to high pressure. A high-performance hot oxygen burner (HOB) was designed and fabricated for use in an existing 1 ton/day pressurized entrained-flow gasifier. Ex-situ testing demonstrated that the HOB provides excellent atomization of the slurries and creates a very uniform and robust flame. Performance of the HOB in the gasifier was exceptional and resulted in better conversion to syngas than any previous generation of burner. Fabricating the HOB for this project was challenging due to the relatively small scale of the gasifier. Performance in larger-scale systems is expected to be even better. Gasification of mixtures of coal, bioliquid and plastic oil proved to be relatively easy. The gasifier was able to be started up, pressurized and transitioned to the blended slurry feedstock with no technical difficulty. Syngas rich in CO and H 2 , with relatively low concentrations of CO 2 , was produced. During startup and at low gasification temperatures, soot formation was observed, as is typical for gasification of petroleum-based feedstocks. No slag buildup was observed under the conditions tested. The project was successful in demonstrating the technical feasibility of the concept. Future development should broaden the range of feedstocks under consideration, more deeply explore performance of the gasifier, and evaluate scale-up and implementation strategies.

08 HYDROGEN↗

Virtual Engineering of Low-Temperature Conversion

In this work, we present the development of an overarching software framework and supporting multiphysics models to simulate the end-to-end process of biomass conversion. This virtual engineering (VE) software is designed with the goal of accelerating research and development and reducing risk for market-relevant biomass conversion processes. We currently support multiple models, computing paradigms, and fidelities representing the steps of feedstock pretreatment, enzymatic hydrolysis, and bioconversion. Although this VE approach was developed to support a biomass workflow, we have designed each component in a way that allows us to easily support new domains, unit models, and feedstocks. We begin by presenting the user-facing aspects of the VE software and highlight how simulated elements are defined and linked by VE functions. We then present an overview of the high-fidelity computational fluid dynamics (CFD) models developed to support our target domain before segueing into our efforts to develop accurate and fast surrogate models, capturing the salient outcomes from the CFD simulations in a significantly less computationally demanding manner. We then present how VE calculations interface with a commercial techno-economic analysis software, Aspen Plus. We conclude by presenting VE case studies that leverage these methods and discuss how our methods can be extended to support a wide variety of accelerated biofuel commercialization pathways in the future.

biofuel↗