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At least 19 records

Natural variation and improved genome annotation of the emerging biofuel crop field pennycress ( Thlaspi arvense )

The Brassicaceae family comprises more than 3,700 species with a diversity of phenotypic characteristics, including seed oil content and composition. Recently, the global interest in Thlaspi arvense L. (pennycress) has grown as the seed oil composition makes it a suitable source for biodiesel and aviation fuel production. However, many wild traits of this species need to be domesticated to make pennycress ideal for cultivation. Molecular breeding and engineering efforts require the availability of an accurate genome sequence of the species. Here, we describe pennycress genome annotation improvements, using a combination of long- and short-read transcriptome data obtained from RNA derived from embryos of 22 accessions, in addition to public genome and gene expression information. Our analysis identified 27,213 protein-coding genes, as well as on average 6,188 biallelic SNPs. In addition, we used the identified SNPs to evaluate the population structure of our accessions. The data from this analysis support that the accession Ames 32872, originally from Armenia, is highly divergent from the other accessions, while the accessions originating from Canada and the United States cluster together. When we evaluated the likely signatures of natural selection from alternative SNPs, we found 7 candidate genes under likely recent positive selection. These genes are enriched with functions related to amino acid metabolism and lipid biosynthesis and highlight possible future targets for crop improvement efforts in pennycress.

59 BASIC BIOLOGICAL SCIENCES↗

Structure and sequence evolution in the pennycress ( Thlaspi arvense ) pangenome

Eukaryotic genomes harbor many forms of variation, including nucleotide diversity and structural polymorphisms, which experience natural selection and contribute to genome evolution and biodiversity. Harnessing this variation for agriculture hinges on our ability to detect, quantify, catalog, and deploy genetic diversity. Here, we explore seven complete genomes of the emerging biofuel crop pennycress ( Thlaspi arvense ) drawn from across the species' current genetic diversity to catalog variation in genome structure and content. Across this new pangenome resource, we find contrasting evolutionary modes in different genomic zones. Gene-poor, repeat-rich pericentromeric regions experience frequent rearrangements, including repeated centromere repositioning. By contrast, conserved gene-dense chromosome arms maintain large-scale synteny across accessions even in fast-evolving NOD-like receptor immune genes, where microsynteny breaks down across species, but gene cluster positioning macrosynteny is maintained. Our findings highlight that multiple elements of the genome experience dynamic evolution that conserves functional content on the chromosome scale but allows repositioning and presence–absence variation on a local scale. This diversity is invisible to classical reference-based strategies and highlights the strength and utility of pangenomic resources. These results provide a valuable case study of rapid genomic structural evolution within a species and powerful resources for crop development in an emerging biofuel crop.

Thlaspi arvense↗

BETO 2021 Peer Review - Analytical Development & Support WBS 2.5.1.101

The objective of the Analytical Development and Support (ADS) Project is to produce and maintain the critical analytical methods and tools that enable evaluation of emerging biofuels R&D at NREL and in the broader biofuels research community. Our project is divided into two tasks: one task to develop novel analytical techniques and improve existing methods and one task to maintain existing analytical capabilities at NREL and provide outreach to the wider community. The ADS Project is world-recognized for our Laboratory Analytical Procedures (LAPS) which provide detailed procedures for compositional analysis of biomass and have been adopted as the de facto standards within the biofuels community largely due to the transparency of the methods and the high reputation of NREL's research. Our dialog with stakeholders allows us to provide robust, precise, accurate, and publicly available analytical procedures for better valuation of scientific tools such as our recent accomplishment in developing a cellulose assay to support the EPA and industry in calculation of converted cellulose during starch ethanol production. We continue to develop analytical capabilities to support BETO's directives to research cost advantaged fuels such as animal wastes and novel bioproducts like 2,3-Butane-diol.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Herbaceous Feedstock 2022 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 in this mission, Idaho National Laboratory completes an annual SOT report for nth-plant and 1st-plant herbaceous biomass feedstock logistics. The purpose of the SOT is to provide the status of feedstock supply system technology development for herbaceous biomass to biofuels relative to technical targets and cost goals from specific design cases, based on data and experimental results. Although conventional feedstock supply systems form the backbone of the emerging biofuels industry, they have limitations that restrict widespread implementation on a national scale. To meet the demands of the future industry, the feedstock supply system must shift from the conventional system to what has been termed “advanced” supply systems. In advanced designs, a distributed network of aggregation and processing centers, termed “depots,” are employed near the points of biomass production (i.e., the field or forest) to reduce feedstock variability and produce feedstocks of a uniform format, moving toward biomass commoditization. The 2022 Herbaceous SOT is part of a vision of achieving an implemented advanced feedstock supply system, which produces a stable, tradable commodity at the decentralized distributed depot. It utilizes feedstock fractionation by incorporating technologies that can separate the biomass into its anatomical fractions (leaves, husks, stems and cobs) to reduce impurities and produce fractions that satisfy downstream quality considerations. By using a series of air classification steps, this strategy can reduce the extrinsic ash in corn stover and produce enriched tissue fractions that can be blended to a conversion specification or converted individually in optimized biochemical conversion campaigns. Additionally, a majority of the leaves (which do not meet the quality specification) are separated out early and can be supplied to alternate markets. The 2022 Herbaceous SOT incorporates an advanced biomass fractionation and processing system to produce pellets enriched tissues from three-pass corn stover. The resulting enriched pellets are delivered to the biorefinery individually where they can be blended to a specification or converted in campaigns where the conditions are optimized for each tissue. Unused fractions can be sent to a a midstream market or to a different conversion process that is better suited to their properties to offset the cost of the delivered feedstock. The main benefits from the proposed system can be summarized as: (1) $6.86/dry ton (2016$) lower cost for the air classification due to elimination of the requirement to discard the high ash lights fraction; (2) $1.56/dry ton lower delivered cost by selling the unsuitable leaf fraction into the feed market as a midstream co-product (assuming a selling price that is 11% higher than their cost of production); (3) 0.98% increase in carbohydrate content (from 60.16% to 61.14%); and (4) 0.97% decrease in ash content (from 6.00% to 5.03%) compared to the 2021 Herbaceous SOT. Overall, the 2022 nth-plant Herbaceous SOT predicts a modeled delivered feedstock cost of $78.64/dry ton (2016$) if it is assumed that the enriched leaf fraction is sold at its production cost; this is a slight increase of $0.43/dry ton increase from the 2021 Herbaceous SOT nth-Supply case cost. The increased cost derived from a $0.38/dry ton increase in transportation and handling cost to procure more biomass (to replace the enriched leaf fraction that was not delivered to the biorefinery. The total preprocessing cost was $0.27/dry ton higher than the 2021 result because of updates to energy consumption, purchasing price and dry matter loss data for the rotary shear ($3.00/dry ton increase) and the pelleting mill ($4.52/dry ton increase). The data utilized were generated in pilot-scale tests in the Biomass Feedstock National User Facility (BFNUF) at INL and at Forest Concepts, including tests for rotary shear and pelleting of the air classified fractions. A greenhouse gas emissions analysis was performed by Argonne National Laboratory using the most up to date version of the Greenhouse Gases, Regulated Emissions, and Energy use in Transportation model (GREET®). The analysis showed an increase of 17.34 kg CO2e/dry ton from the 2021 SOT (67.71 kg CO2e/ton in the 2021 Herbaceous SOT to 85.05 kg CO2e/ton in the 2022 Herbaceous SOT). The net increase is primarily attributed to increased energy consumption in pelleting mill.

09 BIOMASS FUELS↗

Limitations of cetane number to predict transient combustion phenomena in high-pressure fuel sprays

Fundamental understanding of in-cylinder processes in diesel engines is important to screen emerging biofuels and advanced combustion modes that can reduce greenhouse gas emissions and regulated pollutants including soot. In this study, the role of fuel properties on spray development and combustion is investigated by systematically isolating chemical and thermophysical effects. Three different fuels are considered, two with similar chemical properties and two with similar thermophysical properties with one fuel common to both groups. Experiments are performed in a constant-pressure flow chamber de-signed to provide stable test conditions and facilitate acquisition of at least 150 injections in quick succession for each fuel under reacting conditions at high-pressure, high-temperature ambient conditions using a modified conventional diesel engine injector. Further, high speed optical diagnostics including rainbow schlieren deflectometry, OH* chemiluminescence, and a two-color pyrometry system are employed to simultaneously image the transient spray and reacting jets. Image analysis is performed to determine liquid length, vapor penetration length, timing and location of first stage and main ignition events, lift-off location, total soot mass, and more. Results show that fuels with similar chemical properties or cetane number (CN) exhibit similar delay times for first stage and main ignition events as may be expected, but very different liquid length, first stage and main ignition locations, lift-off length, apparent turbulent flame speed, and soot formation. As such, the ability to characterize candidate biofuels with CN or other parameters derived from simple flame configurations is called into question. In this study, thermo-physical properties controlling the liquid length are identified as the main contributing factor for the observed differences.

33 ADVANCED PROPULSION SYSTEMS↗

Identification of microRNAs responsive to arbuscular mycorrhizal fungi in Panicum virgatum (switchgrass)

Background: MicroRNAs (miRNAs) are important post-transcriptional regulators involved in the control of a range of processes, including symbiotic interactions in plants. MiRNA involvement in arbuscular mycorrhizae (AM) symbiosis has been mainly studied in model species, and our study is the first to analyze global miRNA expression in the roots of AM colonized switchgrass ( Panicum virgatum ), an emerging biofuel feedstock. AM symbiosis helps plants gain mineral nutrition from the soil and may enhance switchgrass biomass production on marginal lands. Our goals were to identify miRNAs and their corresponding target genes that are controlling AM symbiosis in switchgrass. Results: Through genome-wide analysis of next-generation miRNA sequencing reads generated from switchgrass roots, we identified 122 mature miRNAs, including 28 novel miRNAs. By comparing miRNA expression profiles of AM-inoculated and control switchgrass roots, we identified 15 AM-responsive miRNAs across lowland accession “Alamo”, upland accession “Dacotah”, and two upland/lowland F 1 hybrids. We used degradome sequencing to identify target genes of the AM-responsive miRNAs revealing targets of miRNAs residing on both K and N subgenomes. Notably, genes involved in copper ion binding were targeted by downregulated miRNAs, while upregulated miRNAs mainly targeted GRAS family transcription factors. Conclusion: Through miRNA analysis and degradome sequencing, we revealed that both upland and lowland switchgrass genotypes as well as upland-lowland hybrids respond to AM by altering miRNA expression. We demonstrated complex GRAS transcription factor regulation by the miR171 family, with some miR171 family members being AM responsive while others remained static. Copper miRNA downregulation was common amongst the genotypes tested and we identified superoxide dismutases and laccases as targets, suggesting that these Cu-miRNAs are likely involved in ROS detoxification and lignin deposition, respectively. Other prominent targets of the Cu miRNAs were blue copper proteins. Overall, the potential effect of AM colonization on lignin deposition pathways in this biofuel crop highlights the importance of considering AM and miRNA in future biofuel crop development strategies.

59 BASIC BIOLOGICAL SCIENCES↗

Temperature Effects on Droplet Oscillation Decay with Application to Fuel Property Measurement

Observation of oscillation decay in droplets has been shown to be an effective approach in determining physical properties such as viscosity and surface tension for emerging biofuels using μl quantities. Herein this work extends the approach to higher temperatures relevant to fuel injection conditions for internal combustion engines. Experiments are conducted that use high-resolution strobed imaging of moving, heated, μm-sized, fuel droplets to capture shape oscillation decay through image processing and analysis. Two fuels are investigated, iso-butanol and a primary reference fuel (PRF 84), which is a mixture of iso-octane and n-heptane. A piezoelectric droplet generator is used to generate a continuous train of single droplets, which are given an initial perturbation and subsequently undergo damped oscillations, captured using strobed imaging. Surface tension and viscosity calculated at four different temperatures ranging from 30°C-55°C using the frequency and decay time associated with the fundamental mode are found to be within ~ 10% of reference values obtained from the literature. Complementary numerical simulations are performed that utilize a volume-of-fluid approach to track transient droplet oscillation phenomena along with heat and mass transfer in a 2D axisymmetric domain. Simulations, where droplet size and temperature can be independently varied, capture an expected decrease in oscillation frequency and increase in decay time, with increase in fuel temperature. The simulations are further used to study the relative contribution to deviations in surface tension and viscosity predictions due to heat and mass transfer from the droplet, as well as viscous effects violating the inviscid flow assumption in the droplet oscillation theory. Mass loss effects are found to be negligible. Temperature change due to heat transfer is found to have the next highest sensitivity, particularly for the more volatile PRF 84, which has a lower viscosity and associated Ohnesorge number. For isobutanol, which has a higher viscosity and Ohnesorge number, viscous effects contribute the most to deviation in fuel property predictions.

09 BIOMASS FUELS↗

Establishing Validation Sites for Field-Level Emissions Quantification from Grain Sorghum in Southern Great Plains

Establish publicly available open-source, high-resolution datasets of greenhouse gas (GHG) emissions, meteorology, soil, and vegetation to support testing and validation of emerging biofuel production monitoring technologies. The project team deployed latest technologies to collect “Gold Standard” data for field- level fluxes of CO 2 , N 2 O, CH 4 and H 2 O in grain sorghum production systems in Texas, Oklahoma, and Kansas. The Oklahoma State University led SMARTFARM was initiated on Oct 1, 2020, due to COVID-19 pandemic delay. We installed closed-path dual laser Eddy Covariance (EC) flux measurement system along with a well-established open-path EC system for simultaneously measuring field-level fluxes of CO 2 , N 2 O, CH 4 , and H2O. The eddy flux data are combined with sub-acre soil moisture and temperature measurements, plant and soil analysis data (nutrient, composition, carbon, growth, and phenology), spatial surveys of soil EC, and pH. Grain sorghum crop was managed including planting and harvesting by producers. All teams recruited postdocs and students for the project and submitted purchase orders for field equipment. The team has developed a draft impact sheet and finalized the advisory board for the project.

09 BIOMASS FUELS↗

Comparative establishment and yield of bioenergy sorghum and maize following pre–emergence waterlogging

Biofuel feedstocks grown on marginal lands, such as those that experience ephemeral waterlogging, reduce interference with food agriculture. As early-season extreme precipitation events in the U.S. Midwest continue to increase, waterlogging tolerance may play an important role in the productivity of annual biofuel cropping systems. We assessed the establishment and yield of photoperiod-sensitive sorghum [Sorghum bicolor (L.) Moench] and maize (Zea mays L.) after extreme early-season rainfall events in central Illinois. We used paired sorghum and maize transects, spanning from low-lying poorly drained areas to higher better drained areas, to evaluate the response of both cropping systems throughout the 2020 growing season. Sorghum maintained 25% mean emergence rates in areas that experienced the most severe waterlogging, but maize failed to establish under the same conditions. Despite the low establishment, sorghum yields in the poorly drained areas were upwards of 50% of those found in the better drained, whereas maize yields were zero areas in the poorly drained locations. The yield compensation in sorghum resulted from increased tillering, higher productivity per stem, and subsequently higher productivity per plant, which illustrates the greater phenotypic plasticity of sorghum compared to maize. Land managers who are currently seeking to grow annual cellulosic feedstocks on soils that are vulnerable to transient soil waterlogging will likely have more success with photoperiod-sensitive bioenergy sorghum than maize.

54 ENVIRONMENTAL SCIENCES↗

Systems Analysis and Engineering of Biofuel Production in Chromochloris zofingiensis , an Emerging Model Green Alga

As a core component of a sustainable bioeconomy, microalgae have the potential to become a major source of biofuels and bioproducts. These photosynthetic microbes utilize solar energy, grow quickly, consume CO 2 , and can be cultivated on non-arable land. However, there are presently considerable practical limitations in the photosynthetic production of biofuels from microalgae, resulting in low productivity and high costs. Algae are a strikingly diverse group and understudied algae can reveal new opportunities for biofuels. Integrative systems biology and engineering of emerging model systems are needed to expand the possibilities of microbial production of biofuels and bioproducts. Our long-term goal is to design and engineer high-level production of biofuel precursors in microalgae.

09 BIOMASS FUELS↗

Production of Biofuels from Biomass by Fungi

The use of abundantly available lignocellulosic biomass as a feedstock for biofuels has emerged as a sustainable alternative to fossil fuels. Industries across the world have shifted their focus to enhancing bioconversion through fungi, efficient and tractable organisms capable of producing valuable cost-effective enzymes. The many capabilities of fungal species offer opportunities for the tailored production of valuable compounds from a variety of substrates. This review details a high level examination of how fungal enzymes degrade plant polysaccharides to simple sugars through the sensitive coordination of transcription factors. Scientists have used a variety of tools like genetically engineering fungi, combining various species, and altering industrial/process conditions to enhance biomass deconstruction and fermentation while solving obstacles like inhibitory compounds. While ethanol and biodiesel are highlighted, additional biofuels are mentioned. These advances have uncovered themes like synergism and biocatalysis, led to the discovery of new fungal species, and revealed unique enzymatic mechanisms. Notably, this research highlights the complexity of metabolic systems within and between fungi. Fungi have been indispensable to the biofuel industry and future research will be crucial for energy sustainability.

Ottum, Eva MN↗

Exploring genetic diversity, population structure, and subgenome differences in the allopolyploid Camelina sativa : implications for future breeding and research studies

Abstract Camelina (Camelina sativa), an allohexaploid species, is an emerging aviation biofuel crop that has been the focus of resurgent interest in recent decades. To guide future breeding and crop improvement efforts, the community requires a deeper comprehension of subgenome dominance, often noted in allopolyploid species, “alongside an understanding of the genetic diversity” and population structure of material present within breeding programs. We conducted population genetic analyses of a C. sativa diversity panel, leveraging a new genome, to estimate nucleotide diversity and population structure, and analyzed for patterns of subgenome expression dominance among different organs. Our analyses confirm that C. sativa has relatively low genetic diversity and show that the SG3 subgenome has substantially lower genetic diversity compared to the other two subgenomes. Despite the low genetic diversity, our analyses identified 13 distinct subpopulations including two distinct wild populations and others putatively representing founders in existing breeding populations. When analyzing for subgenome composition of long non-coding RNAs, which are known to play important roles in (a)biotic stress tolerance, we found that the SG3 subgenome contained significantly more lincRNAs compared to other subgenomes. Similarly, transcriptome analyses revealed that expression dominance of SG3 is not as strong as previously reported and may not be universal across all organ types. From a global analysis, SG3 “was only significant higher expressed” in flower, flower bud, and fruit organs, which is an important discovery given that the crop yield is associated with these organs. Collectively, these results will be valuable for guiding future breeding efforts in camelina.

Agriculture↗

Fueling the Future: The Emergence of Self-Powered Enzymatic Biofuel Cell Biosensors

Self-powered biosensors are innovative devices that can detect and analyze biological or chemical substances without the need for an external power source. These biosensors can convert energy from the surrounding environment or the analyte itself into electrical signals for sensing and data transmission. The self-powered nature of these biosensors offers several advantages, such as portability, autonomy, and reduced waste generation from disposable batteries. They find applications in various fields, including healthcare, environmental monitoring, food safety, and wearable devices. While self-powered biosensors are a promising technology, there are still challenges to address, such as improving energy efficiency, sensitivity, and stability to make them more practical and widely adopted. This review article focuses on exploring the evolving trends in self-powered biosensor design, outlining potential advantages and limitations. With a focal point on enzymatic biofuel cell power generation, this article describes various sensing mechanisms that employ the analyte as substrate or fuel for the biocatalyst’s ability to generate current. Technical aspects of biofuel cells are also examined. Research and development in the field of self-powered biosensors is ongoing, and this review describes promising areas for further exploration within the field, identifying underexplored areas that could benefit from further investigation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The role of social support on midwestern farmers’ willingness to grow perennial bioenergy crops

The lack of farmers' willingness to grow perennial bioenergy crops (PBCs) presents a critical barrier to the emergence of cellulosic biofuel production. The willingness relies on a complex network of economic, environmental, and social drivers, among which the influence of social factors (e.g., the influence of neighborhood, community, and communication) is less understood. This study addresses this knowledge gap via a survey analysis of midwestern farmers. The survey data are analyzed through ordinary least square regression and structural equation model, which together investigate the individual and interactive impacts of multiple factors on farmers' decisions to adopt PBCs. Based on a farm-scale analysis, six statistically significant predictors of farmer willingness to grow PBCs are identified: perception of PBCs' environment benefits, education level, willingness to take risks, familiarity with PBCs, portion of peers already growing PBCs, and support of biorefineries locating in the local community. Among these, the latter three predictors are social support variables. It is found that familiarity with the crops is the most significant predictor of willingness; familiarity is also an important intermediate variable that mediates the influence of many other predictors. In addition, peer adoption can both directly and indirectly affect willingness via its influence on familiarity. Furthermore, these findings suggest that it is a pressing need to improve farmers’ knowledge of PBCs to promote the adoption of such crops.

09 BIOMASS FUELS↗

Ionic liquid-water mixtures enhance pretreatment and anaerobic digestion of agave bagasse

Agave bagasse (AB), a byproduct of tequila production, is a waste generated in large quantities and an emerging alternative for biofuels production. Nevertheless, the use of lignocellulosic biomass for this purpose requires a pretreatment that maximizes its degradation and utilization. Although there are several pretreatment types, using ionic liquids (IL) has become attractive due to its ability to dissolve the lignocellulosic biomass under mild conditions of time and temperature. As a drawback are the high cost of IL's and their high viscosity; thus mixing them with water is a way to overcome these hindrances. This work aimed to evaluate the effect of pretreatment with three diluted ILs on the A. tequilana bagasse structure and sugars production, as well as the potential of the enzymatic hydrolysate to produce methane in batch mode. The evaluated ILs were: 1-ethyl-3-methylimidazolium acetate [Emim][OAc], choline lysinate [Ch][Lys] and ethanolamine acetate [EOA][OAc]. The results showed that the IL with the best performance was [Ch][Lys], which not only offered the highest yields of sugar production (0.57 ± 0.03 g total sugars / g bagasse) but also, it is possible to use it at 30 % in a mix with water obtaining similar yields as with pure IL. Enzymatic hydrolysis of IL pretreated AB achieved 50.7 kg sugars and 49.3 kg of sugars for 90 %-[Ch][Lys] and 30 %-[Ch][Lys], respectively per 100 kg of untreated biomass. The enzymatic hydrolysate from the 30 %-[Ch][Lys] pretreated AB was able to achieve 0.28 L CH4/g CODfed, demonstrating that both sugar and methane yields are maintained when pure or 30 %-[Ch][Lys] are used for pretreatment, representing a step forward towards process feasibility.

59 BASIC BIOLOGICAL SCIENCES↗

Data for The Role of Social Support on Midwestern Farmers’ Willingness to Grow Perennial Bioenergy Crops

The lack of farmers’ willingness to grow perennial bioenergy crops (PBCs) presents a critical barrier to the emergence of cellulosic biofuel production. The willingness relies on a complex network of economic, environmental, and social drivers, among which the influence of social factors (e.g., the influence of neighborhood, community, and communication) is less understood. This study addresses this knowledge gap via a survey analysis of midwestern farmers. The survey data are analyzed through ordinary least square regression and structural equation model, which together investigate the individual and interactive impacts of multiple factors on farmers’ decisions to adopt PBCs. Based on a farm-scale analysis, six statistically significant predictors of farmer willingness to grow PBCs are identified: perception of PBCs’ environment benefits, education level, willingness to take risks, familiarity with PBCs, portion of peers already growing PBCs, and support of biorefineries locating in the local community. Among these, the latter three predictors are social support variables. It is found that familiarity with the crops is the most significant predictor of willingness; familiarity is also an important intermediate variable that mediates the influence of many other predictors. In addition, peer adoption can both directly and indirectly affect willingness via its influence on familiarity. These findings suggest that it is a pressing need to improve farmers’ knowledge of PBCs to promote the adoption of such crops.

Economics↗

The Model System Saccharomyces cerevisiae Versus Emerging Non-Model Yeasts for the Production of Biofuels

Microorganisms are effective platforms for the production of a variety of chemicals including biofuels, commodity chemicals, polymers and other natural products. However, deep cellular understanding is required for improvement of current biofuel cell factories to truly transform the Bioeconomy. Modifications in microbial metabolic pathways and increased resistance to various types of stress caused by the production of these chemicals are crucial in the generation of robust and efficient production hosts. Recent advances in systems and synthetic biology provide new tools for metabolic engineering to design strategies and construct optimal biocatalysts for the sustainable production of desired chemicals, especially in the case of ethanol and fatty acid production. Yeast is an efficient producer of bioethanol and most of the available synthetic biology tools have been developed for the industrial yeast Saccharomyces cerevisiae. Non-conventional yeast systems have several advantageous characteristics that are not easily engineered such as ethanol tolerance, low pH tolerance, thermotolerance, inhibitor tolerance, genetic diversity and so forth. Currently, synthetic biology is still in its initial steps for studies in non-conventional yeasts such as Yarrowia lipolytica, Kluyveromyces marxianus, Issatchenkia orientalis and Pichia pastoris. Therefore, the development and application of advanced synthetic engineering tools must also focus on these underexploited, non-conventional yeast species. Herein, we review the basic synthetic biology tools that can be applied to the standard S. cerevisiae model strain, as well as those that have been developed for non-conventional yeasts. In addition, we will discuss the recent advances employed to develop non-conventional yeast strains that are efficient for the production of a variety of chemicals through the use of metabolic engineering and synthetic biology.

09 BIOMASS FUELS↗

Sustainable Lactic Acid Production from Lignocellulosic Biomass

Lignocellulosic biomass is a promising feedstock for sustainable biofuels and bioproducts. Among emerging bioproducts, lactic acid has attracted significant interest because of its growing application in many industries (e.g., packaging, medical, and pharmaceutical). In this study, BioSTEAM—an open-source platform—was leveraged for the design, simulation, and evaluation (via techno-economic analysis, TEA, and life cycle assessment, LCA) of lignocellulosic lactic acid biorefineries. With a minimum product selling price (MPSP) between 1.38 and 1.91 kg –1 (5th–95th percentiles, baseline at 1.57 kg –1 ), the biorefinery was capable of producing market-competitive lactic acid (market price between 1.7 and 2.1 kg –1 ), and its performance could be further enhanced (e.g., MPSP down to 1.09 kg –1 , global warming potential of 2.79 kg CO 2 -eq·kg –1 , and fossil energy consumption of 31.7 MJ·kg –1 ) with advancements in key technological parameters (fermentation yield and separation process conversions) and optimization in process operation. Sensitivity analyses focused on the fermentation unit (across titer, yield, and productivity; neutral vs low-pH fermentation) and feedstock characteristics (carbohydrate content and price) were also included to quantify their impact on the sustainability of the biorefinery. Overall, this research highlights the ability of agile TEA/LCA to screen promising biorefinery designs, prioritize research needs, and establish a road map for the continued development of bioproducts and biofuels.

09 BIOMASS FUELS↗