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DOE OSTI2022

A tailored fast thioacidolysis method incorporating multi-reaction monitoring mode of GC-MS for higher sensitivity on lignin monomer quantification

Abstract Thioacidolysis is widely used for lignin structural characterization by cleaving β-aryl ethers to release syringyl (S), guaiacyl (G), and p -hydroxyphenyl (H) monomers followed by GC analysis. However, the traditional thioacidolysis method requires tedious extraction steps with chlorinated solvent underlying harmful to health, limiting its efficiency and application. Herein, an improved thioacidolysis method with high sensitivity for the quantitation of lignin-derived monomers was developed. The improved protocol used a quick, streamlined procedure to recover the monomeric products using ethyl acetate as extracting solvent and MS detector in multi-reaction monitoring mode to enhance its ability to detect extremely low concentration (0.1 ppb with signal-to-noise higher than 2) of monomeric products. Additionally, a fast GC program was established to speed up the GC quantitation. Several representative lignocellulose samples, including gymnosperm, angiosperm, and poaceae, were used to test this tailored method. The results demonstrated that the ratios of lignin monomer compositions determined by this method were consistent with that of traditional procedure despite the slightly higher monomer yields measured. More importantly, this method uses non-chlorinated solvent for microscale extraction and requires no evaporation step for workup, which is a green and efficient way for the quantification of lignin monomer compositions.

Forestry↗

DOE OSTI2020

Training Population Optimization for Genomic Selection in Miscanthus

Miscanthus is a perennial grass with potential for lignocellulosic ethanol production. To ensure its utility for this purpose, breeding efforts should focus on increasing genetic diversity of the nothospecies Miscanthus × giganteus (M×g) beyond the single clone used in many programs. Germplasm from the corresponding parental species M. sinensis (Msi) and M. sacchariflorus (Msa) could theoretically be used as training sets for genomic prediction of M×g clones with optimal genomic estimated breeding values for biofuel traits. To this end, we first showed that subpopulation structure makes a substantial contribution to the genomic selection (GS) prediction accuracies within a 538-member diversity panel of predominately Msi individuals and a 598-member diversity panels of Msa individuals. We then assessed the ability of these two diversity panels to train GS models that predict breeding values in an interspecific diploid 216-member M×g F2 panel. Low and negative prediction accuracies were observed when various subsets of the two diversity panels were used to train these GS models. To overcome the drawback of having only one interspecific M×g F2 panel available, we also evaluated prediction accuracies for traits simulated in 50 simulated interspecific M×g F2 panels derived from different sets of Msi and diploid Msa parents. The results revealed that genetic architectures with common causal mutations across Msi and Msa yielded the highest prediction accuracies. Ultimately, these results suggest that the ideal training set should contain the same causal mutations segregating within interspecific M×g populations, and thus efforts should be undertaken to ensure that individuals in the training and validation sets are as closely related as possible.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

DOE Energy Frontier Research Centers Center for Direct Catalytic Conversion of Biomass to Biofuels (C3Bio)

New capabilities to predict, design and control the chemistries of carbon could answer a global imperative to transition from fossil-based to sustainable transportation fuels. While the use of inexpensive hydrocarbons has been an unparalleled achievement and enabler of economic prosperity for many nations, singular dependence upon crude oil has given rise to systemic vulnerabilities in climate, energy, economic, and national security. Lignocellulosic biomass, a renewable and carbon-neutral resource, has the potential to displace an estimated annual equivalent of three billion barrels of oil in the U.S. alone (National Research Council 2009, U. S. Department of Energy, 2011). However, biomass has only one-third the energy density of crude oil (Agrawal and Singh 2009, Richard 2010) and lacks petroleum’s versatility as a feedstock for fuels and chemicals. These limitations keep biomass conversion below the efficiency level needed for strategic impact while the scientific challenge of routing carbon from one molecular context to another remains unmet. In 2009, the Center for Direct Catalytic Conversion of Biomass to Biofuels (C3Bio) recognized the potential of chemical catalysis and fast pyrolysis to overcome such limitations by transforming the main components of biomass (cellulose, xylan, and lignin) from grasses and trees directly to liquid hydrocarbons and aromatic co-products. Enabled by the EFRC high-risk, high-reward approach to grand challenge science, C3Bio researchers have been key national players in disrupting the conventional paradigm of the cellulosic biorefinery into a new future of “no carbon left behind”—the full utilization of carbon from plant cell walls in energy-dense fuels (Fig. 1). We identified catalytic and fast-pyrolytic pathways that utilize cellulose, xylan and, most significantly, lignin. We developed catalytic processes that deoxygenate and transform monomers and isolated polymers into useful products and tested their use with intact biomass. We gained control of lignin synthesis within plants and initiated tailoring biomass to its end-use through the tools of plant molecular biology and genetic engineering. C3Bio breakthroughs have increased the energy density of biomass-derived substrates via catalytic and pyrolytic conversions into products such as benzoquinones, furfural and hydroxymethylfurfural, levoglucosan and levulinic acid, methoxypropylphenols and propylbenzene. Such advances in biomass conversion would not have been possible without simultaneous advances in analytical instrumentation and methodologies, and imaging technologies and applications. The legacy science developed by C3Bio enables design and control strategies for achieving a targeted product portfolio of fuel and chemical feedstocks from a diverse range of native and tailored biomass. Our research provides the knowledge base required for a bio-economy with product streams as diverse in functionality as those of the petrochemical industry. Coupling targeted computational modeling with experimentation, we achieved: (1) fundamental understanding of biopolymers and cell wall architecture assembly, (2) discovery of new chemistries that allow the development of highly selective pathways to fuels and desirable chemicals, and (3) an integrated systems-level understanding to control catalytic and pyrolytic pathways.

09 BIOMASS FUELS↗

DOE OSTI2023

Small Business Voucher Program: Scaling-Up of Bio-Based C5 Building Block Production: Cooperative Research and Development Final Report, CRADA Number CRD-16-00611

Visolis, Inc. is an early stage industrial biotechnology company developing next generation of processes for producing building blocks for bulk polymers and fuels. The company is developing low-cost routes to the chemicals of the future in support of DOE, EERE and BETO goals to grow the U.S. bioeconomy. NREL has capabilities in scale-up of fermentation processes from bench top through pilot scales and also has expertise and technologies related to production of lignocellulosic sugars from cellulosic feedstocks. NREL's Integrated Biorefinery Research Facility (IBRF) houses multiple fermentation vessels/systems ranging in size from 0.5 to 9,000 L along with ancillary capabilities in compositional analysis, separations, and purification. NREL's role will be to scale-up the Visolis process to produce an intermediate product, mevalonic acid, from both dextrose and biomass derived sugars. The goals of the project are to: 1) transfer and reproduce one of Visolis's proprietary fermentation processes at bench top and pilot scale at NREL's IBRF using dextrose as the feedstock, 2) carry out bench-scale fermentations using Visolis's organism and cellulosic sugars, and 3) produce mevalonic acid from the bench top and pilot scale runs for catalyst lifetime testing and conversion to isoprene.

09 BIOMASS FUELS↗

DOE OSTI2020

Woody Feedstocks 2019 State of Technology Report

The U.S. Department of Energy promotes production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement with this overall mission, Idaho National Laboratory completes annual SOT reports for nth-plant biomass feedstock logistics. The purpose of the SOTs is to provide the status of feedstock supply system technology development for biomass to biofuels, based on actual data and experimental results relative to technical targets and cost goals from specific design cases. The 2019 Woody Feedstock SOT presents the State of Technology for feedstock supply to three individual thermochemical conversion pathways that utilize woody feedstocks: Indirect Liquefaction (IDL), Catalytic Fast Pyrolysis (CFP), and Algal-blend Hydrothermal liquefaction (AHTL). The 2019 reactor throat delivered feedstock costs were found to be $63.54/dry ton, $70.15/dry ton and $70.31/dry ton, respectively (2016$).

09 BIOMASS FUELS↗

DOE OSTI2020

Herbaceous Feedstock 2019 State of Technology Report

The U.S. Department of Energy (DOE) promotes the production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the State of Technology (SOT). As part of its involvement with this mission, Idaho National Laboratory (INL) completes an annual SOT report for biomass feedstock logistics. This report summarizes supply system impacts of Bioenergy Technologies Office (BETO)-funded research and development efforts at INL and elsewhere (such as the High-Tonnage Feedstock Logistics projects (Webb et al. 2013a, Webb et al. 2013b, Webb et al. 2013c, Webb and Sokhansanj 2014, Sokhansanj et al. 2014)) that lead to improvements in feedstock supply systems. These include improvements to and observed performance of innovative harvest and collection methods, storage technologies, transportation and handling approaches, and advanced preprocessing technologies. Biomass quality and variability, and the interface between feedstock quality and conversion performance are key drivers in addition to delivered feedstock cost. In this report, we estimate the benefits of R&D improvements to individual supply system unit operations and present the status of feedstock logistics technology development for converting biomass into biofuels. These analyses are supported by experimental data where possible and help to align the SOT relative to the cost goals defined in the Multi-Year Program Plan. The 2019 Herbaceous SOT incorporates several technology changes in feedstock preprocessing and introduces opportunities from the integrated landscape management (ILM) strategy and increased grower participation to reduce biomass access costs, while maintaining or improving grower profitability. During FY18 uneven flow from the horizontal bale grinder was identified as a significant issue limiting preprocessing system throughput. Based on FSL-funded research at INL, the 2019 Herbaceous SOT replaces the horizontal bale grinder used in the first stage size reduction with a bale processor. The improved uniformity of biomass flow entering the PDU eliminated slugging flow from the first stage size reduction and improved the throughput of downstream operations. In order to achieve moisture reduction through frictional heating during grinding (which allowed elimination of the costly rotary drum dryer in previous SOTs), the second stage grinder was changed from a rotary shear, which does not remove moisture, back to a hammer mill. Finally, the 2019 Herbaceous SOT introduces modified three-pass and two-pass corn stover supply curves derived from the BT16 resource assessment, based on FY19 modeling results (WBS 4.2.1.20) quantifying economic benefits of ILM in the supply area, together with modeling results (WBS 1.2.1.5) identifying ILM strategies to increase grower participation. The 2019 Herbaceous SOT report documents the current modeled cost of an herbaceous feedstock supply system from harvest to the pretreatment reactor throat for hydrocarbon fuel production via biochemical conversion, based on equipment and processes now available or potentially available in the near term. The modeled cost also considers both the required quality and the availability of the biomass resources. The 2019 Herbaceous SOT predicts a modeled delivered feedstock cost of $81.37 /dry ton (2016$); this is a $2.30/dry ton (2016$) decrease from the 2018 Herbaceous SOT. Technology improvements that contributed to this modeled cost reduction include reduced cost for the new preprocessing design and quantification of the opportunities of the integrated landscape management (ILM) strategy and an increased grower participation rate to reduce the grower payment portion of biomass access costs, while maintaining or improving grower profitability. A greenhouse gas emissions (GHG) assessment was completed by Argonne National Laboratory using the 2019 Greenhouse Gases, Regulated Emissions, and Energy use in Transportation model, estimating an increase of 14.89 kg CO2e/ton from the 2018 SOT (69.27 kg CO2e/ton in 2018 to 84.16 kg CO2e/ton in 2019). The increase of energy consumption during preprocessing along with higher transportation distance to access low cost biomass from further distance contributed to the increase of GHG emissions in the 2019 Herbaceous SOT. The reason for the increased transportation distances was the cost tradeoff of going farther from the biorefinery to access the cheaper ILM-derived counties (the cheaper price outweighed the cost of increased supply radius).

09 BIOMASS FUELS↗

DOE OSTI2020

Herbaceous Feedstock 2018 State of Technology Report

The U.S. Department of Energy (DOE) promotes the production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement with this mission, Idaho National Laboratory (INL) completes an annual SOT report for biomass feedstock logistics. This report summarizes supply system impacts of Bioenergy Technologies Office (BETO)-funded research and development efforts at INL and elsewhere (such as the High-Tonnage Feedstock Logistics projects (Webb et al. 2013a, Webb et al. 2013b, Webb et al. 2013c, Webb and Sokhansanj 2014, Sokhansanj et al. 2014) that lead to improvements in feedstock supply systems. These include improvements to and observed performance of innovative harvest and collection methods, storage technologies, transportation and handling approaches, and advanced preprocessing technologies. Biomass quality and variability, and the interface between feedstock quality and conversion performance are key drivers in addition to delivered feedstock cost. In this report, we estimate the benefits of R&D improvements to individual supply system unit operations, and present the status of feedstock logistics technology development for converting biomass into biofuels. These analyses are supported by experimental data where possible, and help to align the SOT relative to the cost goals defined in the Multi-Year Program Plan. The 2018 Herbaceous SOT aligned feedstock logistic design with current biorefinery’s design capacity utilized by biochemical conversion platform. Currently biochemical conversion platform utilizes a 725,000 dry ton/year biorefiney design for the techno economic analysis. Hence, feedstock delivered cost in the 2018 Herbaceous SOT is calculated based on biorefinery’s 725,000 dry ton design capacity instead of 800, 000 dry ton capacity utilized in the 2017 Herbaceous SOT. Biomass availabilities in this SOT were updated to year 2018 data from the 2016 Billion-Ton Report (BT16) (DOE 2016a), with the exception of switchgrass, for which the 2018 Herbaceous SOT utilized the 2019 switchgrass availability data from BT16. The BT16 report (DOE 2016a) does not project switchgrass availability in 2018; the soonest switchgrass is available in the BT16 report is 2019. Therefore, availability of switchgrass for this analysis was that projected for 2019. The 2018 Herbaceous SOT incorporates same technologies utilized in the 2017 Herbaceous SOT. However, a sensitivity analysis is performed to understand the impact of variation of process parameters on those technologies on feedstock logistic cost. New R&D data that shows the variations of process parameters affecting process performance is incorporated in the 2018 SOT to measure the variations in delivered feedstock cost. The 2018 Herbaceous SOT has also provided projected delivered feedstock of 2022 design case based on near term technical target under BETO funded R&D project. Finally, updated biorefinery size of 725,000 dry ton/year was incorporated within least-cost formulation model to select optimal siting and depot scales during optimization of the least cost blend. This modification to the optimization algorithm allows the trade-off between the cost of increased supply radius and the savings from selecting biomass from higher producing counties to be assessed. Such optimization has also showed the economic benefit of decentralized depots in comparison to centralized preprocessing co-located with the biorefinery by decoupling the biorefinery and feedstock locations. The 2018 Herbaceous SOT report documents the current modeled cost of a herbaceous feedstock supply system (from harvest to the pretreatment reactor throat, including grower payment) for hydrocarbon fuel production via biochemical conversion, based on equipment and processes now available or potentially available in the near term. The modeled cost also considers both the required quality and the availability of the biomass resources. The 2018 Herbaceous SOT predicts a modeled delivered feedstock cost of $83.67/dry ton (2016$); this is a $0.23/dry ton (2016$) decrease from the 2017 Herbaceous SOT. The modification of biorefinery’s designed capacity and increased projected biomass availability in the same supply shed contributed to this modeled cost reduction. The least-cost formulation model to optimally site and scale local distributed preprocessing depots also contributed to the cost reduction by considering county-level grower payment and distance from the biorefinery as variables in the optimization algorithm. Sensitivity analysis on various process parameters that affect delivered feedstock cost in the 2018 Herbaceous SOT shows that the delivered cost could varies from $80.45-$88.83/dry ton. The top factors that causes such variations are: effective baling rate, bale density, hammer mill throughput, interest rate and storage dry matter loss.

09 BIOMASS FUELS↗

DOE OSTI2020

Conceptual Basis and Techno-Economic Modeling for Integrated Algal Biorefinery Conversion of Microalgae to Fuels and Products (2019 NREL TEA Update: Highlighting Paths to Future Cost Goals via a New Pathway for Combined Algal Processing)

The report documents the conceptual basis for a new potential Combined Algal Processing design strategy which may allow more flexibility in accommodating different algal biomass feedstock compositions, by enabling upgrading of both protein and carbohydrates in a single step, without a strict requirement for either component to be in soluble or monomeric form, while maintaining effective wet lipid extraction techniques to enable high lipid recoveries. In light of previously-established constraints around algal biomass costs (which are significantly higher than lignocellulosic terrestrial biomass), the present CAP processing strategy reflects an integrated biorefinery concept producing both fuels and value-added chemical coproducts as a means to improve profitability and generate coproduct revenues to help drive down the minimum fuel selling price (MFSP) towards economically viable levels. Namely, this report highlights an integrated CAP biorefinery process and associated technical targets that would be required to achieve U.S. Department of Energy target MFSP goals of $2.5/gallon gasoline equivalent by 2030. This is accomplished by a process involving low-cost seasonal storage of algal biomass during high-growth seasons, rapid flash hydrolysis pretreatment of the biomass, solvent extraction of pretreated biomass, cleanup and fractionation of lipids into triglyceride and free fatty acid fractions, and a series of thermochemical conversion steps to upgrade carbohydrates and protein to hydrocarbon fuels. These steps include mild oxidative treatment (MOT), a process originally investigated at NREL for upgrading lignin, followed by catalytic ketonization and hydrotreating of MOT products to fuels. Isolated triglycerides are sent to a coproduct train, with the base case focused on upgrading to polyurethane foams as a high-value, high-market-volume coproduct.

09 BIOMASS FUELS↗

DOE OSTI2020

Woody Feedstocks 2020 State of Technology Report

The U.S. Department of Energy promotes production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement with this overall mission, Idaho National Laboratory completes annual SOT reports for nth-plant biomass feedstock logistics. The purpose of the SOTs is to provide the status of feedstock supply system technology development for biomass to biofuels, based on actual data and experimental results relative to technical targets and cost goals from specific design cases. The 2020 Woody Feedstock SOT is separated into three separate pathways that utilize woody feedstocks: indirect liquefaction (IDL), catalytic fast pyrolysis (CFP), and algal-blend hydrothermal liquefaction (AHTL).

09 BIOMASS FUELS↗

DOE OSTI2021

Decontamination and Sterilization of Surfaces using Low Cost Ionic Liquids (CRADA Final Report)

The current study made a comprehensive literature review of the structure activity relationship (SAR) for microcidal and virucidal influence of ionic liquids. It summarized the key structure parameters that have close correlation to the antimicrobial and antiviral properties of the ILs. It proposed an experimental plan on developing and testing an effective ionic liquid from renewable lignocellulosic biomass. The goal is to understand the structure activity relationship (SAR) for virucidal influence of ionic liquids through comprehensive literature search and data analysis. A full experimental plan will be drafted based on the literature summary. Perform a comprehensive literature search to aid the development of an effective and easily deployable method to inactivate SARS-CoV-2 and decontaminate surfaces based on low-cost functional ionic liquids. These designer solvents have the potential to be a compelling and environmentally friendly alternative to common chemical disinfectants. Literature summary report including reported ionic liquid structures, synthesis and testing methods, and the structure activity relationship. Experimental plan on developing a low cost effective antiviral ionic liquid.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Elemental Analysis of Bio-Oils by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Laboratory Analytical Procedure (LAP), Issue Date: May 13, 2022

Concentrations of inorganic elements is a key quality metric for bio-oils as certain elements impact upgrading processes and product quality. Unless reduced or removed during production and processing, alkali and alkaline metals native to lignocellulosic biomass can carry over into bio-oils contributing to ash content and degraded catalyst performance during upgrading to hydrocarbon fuels or chemical products. Non-metallic elements such as sulfur and phosphorus can also negatively impact upgrading catalysts and product quality. Inductively coupled plasma optical emission spectroscopy (ICP-OES) can be used to measure inorganic elements of interest in bio-oils. This procedure covers the preparation and analysis of fast pyrolysis (FP) and catalytic fast pyrolysis (CFP) bio-oils. The concentrations of these elements can indicate potential bio-oil quality from the perspective of deoxygenation processes. The implications of elemental composition will depend on process parameters such as upgrading catalyst sensitivities. This Laboratory Analytical Procedure (LAP) covers two methods for quantification of inorganic elements by ICP-OES: Procedure A uses microwave assisted digestion with concentrated nitric acid, and Procedure B is an organic ICP-OES method utilizing a diglyme solvent.

09 BIOMASS FUELS↗

DOE OSTI2022

Synthetic Microbial Consortium for Biological Breakdown and Conversion of Lignin

The plant polymer lignin is the most abundant renewable source of aromatics on the planet and conversion of it to valuable fuels and chemicals is critical to the economic viability of a lignocellulosic biofuels industry and to meeting the DOE’s 2022 goal of $\$2.50$/gallon mean biofuel selling price. Presently, there is no efficient way of converting lignin into valuable commodities. Current biological approaches require mixtures of expensive ligninolytic enzymes and engineered microbes. This project was aimed at circumventing these problems by discovering commensal relationships among fungi and bacteria involved in biological lignin utilization and using this knowledge to engineer microbial communities capable of converting lignin into renewable fuels and chemicals. Essentially, we aimed to learn from, mimic and improve on nature. We discovered fungi that synergistically work together to degrade lignin, engineered fungal systems to increase expression of the required enzymes and engineered organisms to produce products such as biodegradable plastics precursors.

09 BIOMASS FUELS↗

DOE OSTI2022

TCF High Efficiency Anaerobic Electroporation

09 BIOMASS FUELS↗

DOE OSTI2023

Metaoptics Enabled Multifunctional Imaging

Despite the fact that most microorganisms occupy two- and three-dimensional space in heterogeneous arrangements and in proximity to other microorganisms of different species, much of our current knowledge about their metabolic processes is derived from homogenizing, then analyzing, the chemical content of cultures containing a single microbial species. This project addressed this fundamental dichotomy by developing a set of optical imaging principles combining multiple new technologies. The imaging strategies developed in this project combine metaoptics structures with active in situ nanoscale control of the chemical environment and applied them to a microbial system, Myxococcus xanthus, with particular relevance to the DoE mission. The combination of metaoptical architectures and nanoscale control over the molecular environment enables: (a) precise control over the electromagnetic (EM) field at length scales smaller than the wavelength of light; (b) control of the interaction of the EM field with critical molecular systems in DOE relevant microbes; (c) control over the chemical environment – especially the presence and quantity of reactive oxygen species (ROS) that can affect redox homeostasis; and (d) the ability to ask new kinds of questions not accessible to ‘omics’ approaches or standard methods of biological imaging. These capabilities are applicable to detailed studies of metabolic pathways in microbes and to lignocellulosic biomass deconstruction. To accomplish these objectives, we pursued two over-arching technical goals: (1) the development of new metaoptics-enabled approaches to imaging and spectroscopic characterization; and (2) the development of tools to control the chemical environment of a microbial sample with nanometer-scale precision. Goal 1 was addressed through the design, fabrication, and characterization of new metasurfaces capable of super-resolution imaging through extreme confinement of the optical field. Goal 2 was addressed by controlling the redox potential on the nanoscale in microbial communities and characterizing their effect on intrinsic bacterial fluorophores which act as molecular sentinels and through characterization of soluble factors secreted by Myxococcus xanthus by confocal Raman imaging. The optical imaging/sensing approaches developed here make it possible to use these powerful new imaging and sensing modalities in metabolic studies by making it possible to visualize and track the spatial and temporal expression patterns of natural or engineered pathways in microorganisms.

09 BIOMASS FUELS↗

DOE OSTI2022

Syntrophic Co-Cultures of Clostridium Organisms to Produce Higher Alcohols & Other C6-C8 Metabolites (Final Report)

The goal of this project was to advance the systems biology understanding and predictive modeling of synthetic & syntrophic Clostridium microbial consortia, focusing on elucidation of metabolic networks and environmental signals in the consortia. The project has direct applicability to lignocellulosic-biomass based production of higher alcohols as advanced biofuels and C6-C8 metabolites, that can be used as chemicals or serve as biofuel precursors. This project studied and optimized three synthetic syntrophic systems. It examined the population dynamics using flow-cytometry, time-lapse microscopy and PCR analysis. 13C-based tracer analyses was used to examine the metabolite exchange between the syntrophic cell populations and the impact of those interactions on the transcriptome of the individual populations. To enhance our analytical and predictive capabilities, genome-scale models (GSMs) for these syntrophies was developed. RNAseq data for these syntrophic coculture systems were acquired to enable a molecular level understanding of the syntrophies aiming to identify the genetic networks of each organism in the co-culture and compare those against the networks of pure cultures.

09 BIOMASS FUELS↗

DOE OSTI2021

Woody Feedstocks 2021 State of Technology Report

09 BIOMASS FUELS↗

DOE OSTI2021

Herbaceous Feedstock 2021 State of Technology Report

09 BIOMASS FUELS↗

DOE OSTI2023

TCF High Efficiency Anaerobic Electroporation (CRADA Final Report)

The Joint BioEnergy Institute (JBEI) researchers were co-inventors of the technology that will be used on this project and have developed a more current version of the chip and controller. JBEI will also assist with the design of the pathways and implementation of pathways on the chip. LanzaTech has developed novel gas fermentation technology that captures and utilizes greenhouse gases for production of fuels and chemicals. In contrast to traditional fermentation that uses sugars as substrate (and releases CO2 as a byproduct), gas fermentation utilizes C1 substrates carbon monoxide (CO) or CO2. This enables a diverse range of feedstock options including waste gases from industrial sources (e.g., steel mills and processing plants) or syngas generated from any biomass resource (e.g., agricultural waste, municipal solid waste, or organic industrial waste). Biomass is then gasified, allowing for maximum yields and complete carbon utilization including the recalcitrant lignocellulosic fraction that cannot be utilized in traditional sugar fermentation. To maximize the value that can be added to the array of gas resources that the LanzaTech process can use as an input, LanzaTech has pioneered genetic modification of acetogens and developed a comprehensive set of genetic tools to perform routine strain modification, including genome editing tools as CRISPR/Cas9 and libraries of validated genetic parts as promoters and terminators. Using this platform, production of over 50 new molecules have been demonstrated directly from gas. For a few selected molecules production rates and yields have been optimized and surpass production of native producers and engineered E. coli or yeast strains, but a higher throughput approach for combinatorial optimization of pathways is required to further advance synthesis of additional products in parallel.

09 BIOMASS FUELS↗