166 Search Results

Advancing lignocellulose biorefining is imperative for the deployment of cellulosic (2G) biofuels. This work investigates the tailoring of the alkaline deacetylation and mechanical refining (DMR) pathway for the bioconversion of sugarcane bagasse. Experiments are conducted at laboratory and pilot scales, varying the pretreatment conditions (70-92 degrees C; 48-100 gNaOH/kg) and the mechanical refining technologies (PFI and disk refining). The pretreatments selectively solubilize acetyl groups (> 86%) and lignin (10-63%) while mostly preserving structural carbohydrates in the solid phase. Enzymatic hydrolysis generates hydrolysates of clean sugars (glucose and xylose), with sugar yields increasing up to 81% for glucose and 89% formore » xylose in response to delignification and mechanical refining. Biochemical methane potential assays reveal specific methane productions of up to 568 NmL CH4 gVS-1 for alkaline liquor monodigestion and 344 NmL CH4 gVS-1 for co-digestion with sugarcane vinasse from the conventional (1G) sugarcane ethanol, indicating a strong potential for bioenergy recovery from this process stream. Synergies are identified in integrating 1G ethanol, 2GDMR processing of bagasse, and anaerobic co-digestion of 1G vinasse and 2GDMR alkaline liquor. This technology enables sugarcane biorefineries to enhance the co-production of ethanol, methane, and concentrated streams of CO2.« less

Metabolic engineering of vegetative tissues for lipid production holds transformative potential for sustainable biofuels, yet achieving sufficient yields remains challenging. Here, we present a strategy to enhance fatty acid synthesis by redirecting cytosolic phosphoenolpyruvate (PEP) into plastids through overexpression of the plastidial phosphoenolpyruvate/phosphate translocator (PPT1) in vegetative tissues of Arabidopsis thaliana. Integrated metabolomic and transcriptomic analyses revealed that AtPPT1 overexpression alleviated metabolite overaccumulation in high-sugar tissues, consistent with enhanced carbon flux coordination between the cytosol and chloroplast. Notably, phosphofructokinase activity, a key step in glycolysis, was elevated, linking plastidic PEP import to increased glycolytic throughput. In Arabidopsis, overexpression of AtPPT1more » increased fatty acid content and lipid droplet accumulation in the sugar-accumulating sweet11;12;13 mutant, but not in wild-type Col-0. Together, these findings establish PEP redirection as an effective strategy to boost fatty acid and lipid production in sugar-rich vegetative tissues and provide a complementary metabolic module for future lipid-engineering efforts.« less

Switchgrass (Panicum virgatum) is a promising bioenergy crop due in part to its resilience to drought stress. However, the significance of drought timing remains poorly understood, both from a plant biology perspective and its impact on downstream biofuel production. This study determines the developmental stage-specific physiological and metabolic responses of switchgrass to drought stress and its implications for biofuel production using a custom-built programmable irrigation system. Vegetative, flowering, and senescence-stage drought significantly reduced carbon dioxide assimilation, and stomatal conductance without affecting biomass yield. Metabolic profiling revealed significant accumulation of glucose, fructose, quinic acid, shikimate and GABA during vegetative-stage drought, whilemore » flowering and senescence stages exhibited limited metabolic changes. Similarly, specialized metabolites also displayed distinct developmental patterns, with vegetative-stage drought driving the most pronounced metabolic alterations. Thermochemically-treated and hydrolyzed switchgrass biomass from vegetative-stage drought showed elevated lignocellulose-derived compounds and saponins with the latter most positively correlating with fermentation lag times. Conversely, senescence-stage drought enhanced ethanol yields while lowering saponin levels in the hydrolysates. While vegetative-stage drought enhanced physiological resilience, it compromises downstream biofuel production by introducing fermentation inhibitors, particularly saponins.« less

Synthetic biology generates vast combinatorial designs, yet high-throughput analytical methods to screen them are poorly matched to interrogate this search space. We address this challenge by developing a biosensor-driven, growth-coupled selection strategy in Pseudomonas putida for isoprenol, a potential aviation fuel precursor. We found and characterized a noncanonical signaling pathway, revealing a functional and physical complex between a hybrid histidine kinase and an alcohol dehydrogenase, whose activity is tuned by heterodimerization. Leveraging this biosensor in a pooled CRISPRi library selection, we identified key host limitations. Iterative combinatorial strain engineering derived from these hits yielded a 36-fold titer increase to ~900more » milligrams per liter. Integrated omics analysis revealed that metabolic rewiring toward amino acid catabolism was crucial for this improvement. This observation was found to be beneficial by technoeconomic analysis. Our modular workflow provides a powerful strategy for optimizing complex heterologous pathways and uncovering emergent host biology.« less

Freight rail is more energy-efficient than truck transport over long-haul distances, offering a low-energy and emissions-intensive option for transporting freight. This study evaluates techno-economic analysis and life cycle assessment of seven alternative unblended fuels for freight locomotive engines─biodiesel, renewable diesel (RD), bio-oils, methanol, dimethyl ether (DME), ethanol, and ammonia─across 16 fuel pathways utilizing soybean, corn, woody biomass, renewable hydrogen, and waste sources, e.g., sludge, manure, and industrial CO₂, and compares these to conventional diesel. The minimum fuel selling price (MFSP) ranged from $$\$$2.05$ to $$\$$8.27$ per diesel gallon equivalent (2020 US dollars), with biocrude and RDs produced from hydrothermal liquefactionmore » (HTL) of sludge having the lowest MFSPs due to coproduct credits and avoided waste treatment cost. Life cycle GHG emissions ranged from −41 to 53 g of CO₂e/MJ. RD from waste via HTL achieves negative emissions by diverting sludge/manure from GHG-intensive conventional management. Few pathways such as biocrude, methanol, and DME require additional control for SO_X emissions in the refinery, while ethanol, FT-diesel, and bio-oil require additional control for particulate matter emissions. Bio-oil and RD from sludge have lower marginal abatement cost or MAC (–$$\$$38$/tonne CO₂ lowest) while methanol and ammonia with renewable hydrogen have higher MAC ($$\$$490$/tonne CO₂ maximum).« less

In coevolving species, parasites locally adapt to host populations as hosts locally adapt to resist parasites. Parasites often outpace host local adaptation since they have rapid life cycles, but host diversity, the strength of selection, and external environmental influence can result in complex outcomes. To better understand local adaptation in host–parasite systems, we examined locally adapted switchgrass (Panicum virgatum), and its leaf rust pathogen (Puccinia novopanici) across a latitudinal range in North America. We grew switchgrass genotypes in 10 replicated multiyear common gardens, measuring rust severity from natural infection in a ‘host reciprocal transplant’ framework for testing local adaptation. Wemore » conducted genome-wide association mapping to identify genetic loci associated with rust severity. Genetically differentiated rust populations were locally adapted to northern and southern switchgrass, despite host local adaptation to environmental conditions in the same regions. Rust resistance was polygenic, and distinct loci were associated with rust severity in the north and south. We narrowed a previously identified large-effect quantitative trait locus for rust severity to a candidate YELLOW STRIPE-LIKE gene and linked numerous other loci to defense-related genes. Overall, our results suggest that both hosts and parasites can be simultaneously locally adapted, especially when parasites impose less selection than other environmental factors.« less

Biofuel production from lignocellulosic biomass offers a transformative solution to reduce global fossil fuel dependency. Certain thermophilic anaerobes, including Clostridium thermocellum, show promise for renewable ethanol production due to their ability to break down plant material at high temperatures. However, achieving commercially viable ethanol yields has proven challenging despite extensive engineering efforts. Here, we characterized 27 ferredoxin:NADP⁺ oxidoreductase (Fnor) enzymes for their enzyme activity, nicotinamide cofactor specificity, thermotolerance, and functional expression in C. thermocellum. We identified a subset of 10 of these enzymes as a novel class of Fnor enzymes suited for metabolic pathways aimed at high-titer ethanol production. Whenmore » expressed in engineered C. thermocellum, these enzymes increased ethanol production up to 2.2-fold. These findings establish a novel ethanol pathway and provide insights into physiological roles and biotechnological applications of this new class of Fnor enzymes.« less

The relationship between precipitation and evapotranspiration (ET) is critical to understanding water cycle related dynamics in ecosystems, including crops. Existing studies of bioenergy crops have primarily focused on annual or seasonal ET rates, with less attention given to the immediate ET response following precipitation events. This study examines the variation in ET rates in the days subsequent to precipitation events across various bioenergy crops—corn, switchgrass, and prairies—utilizing 13 years (2010–2022) of growing season data. Meteorological and eddy covariance flux data were collected from seven eddy covariance flux towers as part of the GLBRC scale-up experiment at the Kellogg Biological Stationmore » Long Term Ecological Research sites. The analysis revealed that average ET peaked the day after precipitation and declined linearly over the following days, with a statistically significant relationship (p-value = 0.00027, R2 = 0.96). Neither the type of biofuel vegetation nor the historical land use significantly influenced ET post-precipitation events (p-values = 0.53 and 0.153, respectively). Key predictors of ET following precipitation events include shortwave radiation, season, day of the year, ambient temperature, vapor pressure deficit (VPD), long-wave radiation, precipitation amount, soil moisture, and annual variability. These findings enhance our comprehension of ET responses in bioenergy crop systems, with implications for water management in sustainable agriculture.« less

The biomass-based platform molecule 2,3-butanediol (2,3-BDO) has a wide range of applications in production of sustainable fuels, chemicals, synthetic rubber, and others. However, the selective separation of 2,3-BDO from multicomponent fermentation broths presents challenges due to its low concentration, high solubility in water, high boiling point, and presence of many other species. Here, we demonstrate remarkably selective enrichment and recovery of 2,3-BDO from a corn stover hydrolysate fermentation broth by a pure-silica nano-MFI-type zeolite adsorbent. By means of cyclic and simulated moving bed adsorption processes, we obtained concentrated aqueous 2,3-BDO streams from the fermentation process stream with ∼93% purity andmore » 3-fold enrichment, and >98% purity and 8-fold enrichment, respectively. These findings provide strong support for large-scale adsorptive separation for biobased 2,3-BDO production.« less

While crop and grassland usage continues to increase, the full diversity of plant-specific volatile organic compounds (VOCs) emitted from these ecosystems, including their implications for atmospheric chemistry and carbon cycling, remains poorly understood. It is particularly important to investigate VOCs in the context of potential biofuels: aside from the implications of largescale land use, harvest may shift both the flux and speciation of emitted VOCs. To this point, we evaluate the diversity of VOCs emitted both pre and postharvest from “Alkar” tall wheatgrass (Thinopyrum ponticum), a candidate biofuel that exhibits greater tolerance to frost and saline land compared to othermore » grass varieties. Mature plants grown under field conditions (n = 6) were sampled for VOCs both pre- and postharvest (October 2022). Via hierarchical clustering of emitted VOCs from each plant, we observe distinct “volatilomes” (diversity of VOCs) specific to the pre- and postharvest conditions despite plant-to-plant variability. In total, 50 VOCs were found to be unique to the postharvest tall wheatgrass volatilome, and these unique VOCs constituted a significant portion (26%) of total postharvest signal. While green leaf volatiles (GLVs) dominate the speciation of postharvest emissions (e.g., 54% of unique postharvest VOC signal was due to 1-penten-3-ol), we demonstrate novel postharvest VOCs from tall wheatgrass that are under characterized in the context of carbon cycling and atmospheric chemistry (e.g., 3-octanone). Continuing evaluations will quantitatively investigate tall wheatgrass VOC fluxes, better informing the feasibility and environmental impact of tall wheatgrass as a biofuel.« less