A bacterial toolkit for plants
Here, we highlight efforts to engineer microorganisms and their plant hosts to address the challenges of sustainable agriculture.
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Here, we highlight efforts to engineer microorganisms and their plant hosts to address the challenges of sustainable agriculture.
The Artemis Program mandate for sustainable, long-term presence on the Moon requires learning to "Live Off the Land". Geologic maps enable a paradigm shift to model the development and scaling of infrastructure for robotic and human exploration inside a bounded system on the lunar surface. Relying on geologic maps produced from orbital data and robotic precursor missions, in-situ resource utilization (ISRU) will be required to produce infrastructure, building materials and consumables. The ISRU value chain must be mapped to identify linkages of resources, producers, processes and equipment capabilities for transforming excavated regolith into usable feedstocks. Coordination will be required to balance supply and demand across the whole system of lunar infrastructure and operations, at each stage of development.
Abiotic stresses can cause significant damage to plants. For sustainable bioenergy crop production, it is critical to generate resistant crops to such stress. Engineering promoters to control the precise expression of stress resistance genes is a very effective way to address the problem. Here we developed stably transformed Populus tremula × Populus alba hybrid poplar (INRA 717-1B4) containing one-of-six s ynthetic d rought stress-inducible promoters (SDs; SD9-1, SD9-2, SD9-3, SD13-1, SD18-1, and SD18-3) identified previously by transient transformation assays. We screened green fluorescent protein (GFP) induction in poplar under osmotic stress conditions. Of six transgenic lines containing synthetic promoter, three lines (SD18-1, 9-2, and 9-3) had significant GFP expression in both salt and osmotic stress treatments. Each synthetic promoter employed heptamerized repeats of specific and short cis -regulatory elements (7 repeats of 7-8 bases). To verify whether the repeats of longer sequences can improve osmotic stress responsiveness, a transgenic poplar containing the synthetic promoter of the heptamerized entire SD9 motif (20 bases, containing all partial SD9 motifs) was generated and measured for GFP induction under osmotic stress. The heptamerized entire SD9 motif did not result in higher GFP expression than the shorter promoters consisting of heptamerized SD9-1, 9-2, and 9-3 (partial SD9) motifs. This result indicates that shorter synthetic promoters (~50 bp) can be used for versatile control of gene expression in transgenic poplar. These synthetic promoters will be useful tools to engineer stress-resilient bioenergy tree crops in the future.
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Our reliance on agriculture for sustenance, healthcare, and resources has been essential since the dawn of civilization. However, traditional agricultural practices are no longer adequate to meet the demands of a burgeoning population amidst climate-driven agricultural challenges. Microalgae emerge as a beacon of hope, offering a sustainable and renewable source of food, animal feed, and energy. Their rapid growth rates, adaptability to non-arable land and non-potable water, and diverse bioproduct range, encompassing biofuels and nutraceuticals, position them as a cornerstone of future resource management. Furthermore, microalgae’s ability to capture carbon aligns with environmental conservation goals. While microalgae offers significant benefits, obstacles in cost-effective biomass production persist, which curtails broader application. This review examines microalgae compared to other host platforms, highlighting current innovative approaches aimed at overcoming existing barriers. These approaches include a range of techniques, from gene editing, synthetic promoters, and mutagenesis to selective breeding and metabolic engineering through transcription factors.
The inefficient management of industrial waste threatens environmental sustainability, requiring scalable resource recovery solutions. To address these challenges, we propose a novel and sustainable in-situ one-step strategy for the simultaneous recovery and functionalization of lignin from industrial black liquor, using a sustainable organic acid system assisted by choline chloride. The functionalized lignin adsorbent material (FLAM) exhibited superior adsorption performance, effectively treating animal feedlot wastewater to recover ammonia. The ammonia enriched FLAM was subsequently repurposed as a slow-release fertilizer, enhancing plant growth and demonstrating a closed-loop pathway for waste valorization and nutrient recycling. Methylene blue was used as a model cationic compound to simulate the adsorption behavior and interactions. FLAM achieved a high adsorption capacity of 725.98 mg/g across a broad pH range, significantly outperforming commercial kraft lignin. This sustainable approach advances circular economy principles by integrating remediation and resource recovery, offering a scalable, eco-friendly solution for environmental applications.
The HyTEC Project goal is to mature compact core aircraft engine technologies as part of the NASA-led Sustainable Flight National Partnership (SFNP) that contributes to the United States Aviation Climate Action Plan goal for aviation net zero carbon emissions by 2050. Individual technologies were proposed and defined by industry partners, and then selected technology maturation activities are cost shared. Phase 1 contracts were awarded in 2021, and since then, the project has matured numerous technologies to TRL 4-5. Some of the Phase 1 efforts have been completed with successful results. An initial systems analysis study has been completed to estimate the impact of Phase 1 technologies against project performance metrics, with results indicating significant progress is being made toward meeting those metrics. HyTEC will culminate with an engine core demonstration that will integrate many of the Phase 1 technologies into a large-scale integrated ground demonstration that will achieve TRL 6 to enable industry to transition the technologies into the next single-aisle engine architecture. The goal of the demonstration is to fully meet the project performance metrics that define a compact engine core with substantial improvements in efficiency, durability, performance, hybridization, and sustainability over the baseline established in the year 2020. HyTEC will also incorporate hybrid electric technology into the engine core with the intention to bring the first electrified turbofan engines into production. The core demonstration has been awarded with a cost-share partnership and is planned to occur by the end of 2028.
Understanding the characteristics of travelers and situations that are more likely to switch from single occupancy vehicles (SOV) to more sustainable mobility options can help transportation agencies develop better behavior change strategies. Existing mode shift programs, however, rarely identify their target audience, which could make the programs less effective. Here, to investigate the demographics and trip characteristics of those who are more likely to shift from SOV to carpooling, public transit, and micromobility (e.g. walking, biking), a matrix decomposition audience segmentation method is proposed and applied on travel survey data. The results of this research show that for short-distance leisure or social trips, young and middle-income people are more likely to shift to these mobility options from SOV. This study provides a comprehensive and profound understanding of the characteristics of the target audience that can inform policies to create more targeted behavior change strategies to reach their sustainability goals.
The testing of the combustion stability characteristics of Space Shuttle Main Engine (SSME) 0208 is described in terms of augmenting the technology base for large O/H thrust-chamber assemblies. The throat area is increased by 12 percent over that of the flight SSMEs, and the thrust chamber assembly does not include stability aids. Acoustic modes in the chamber are excited by means of rapid pressure generators employed in the start-transient through mainstage operations. Stability characteristics are determined by damp times which are facilitated by high-frequency instrumentation measuring oscillations and locating stable operating regions. All vibration modes are damped to within the requirements for a chamber mode set forth by the Chemical Propulsion Information Agency. No sustained chamber acoustic oscillations are exhibited in engine 0208's combustion chamber configuration in spite of the absence of baffles and acoustic cavities.
The SUbsonic Single Aft eNgine (SUSAN) program is being developed as a sustainable subsonic regional aircraft. NASA is conducting trade studies for this future hybrid single aisle passenger aircraft. The goal of reducing emission levels by 50% will be enabled through an advanced design. An aft mounted single turbine engine provides thrust and electric power to drive 16 wing mounted electric engines. Using the 16 counter-rotating electric fans will create an efficient, very high bypass ratio, engine. Electric engines allow for distributed thrust, reducing control surfaces and provide a greater dynamic response. Batteries provide boost for takeoff and climb, allowing a smaller, more efficient, turbine engine. As part of the SUSAN effort, two pilot-in-the-loop studies were conducted at NASA Langley Research Center in Virginia to explore flight deck design issues. In particular: how many throttles are required/desired for a 17-engine aircraft, does the hybrid electric design change the answer, what are the requirements for electric engine displays, what system pages are required, and what information is required to monitor the batteries. Objective and subjective data were collected after each scenario and at the end of the day with a guided set of questionnaires from sixteen crews paired by airline. Data along with pilot comments were used to improve the throttle and engine displays between the studies and used to make the final recommendations on flight deck configurations. This paper provides final design recommendations for the SUSAN flight deck and justification for each recommendation.
With increasing concerns about global warming, the push for sustainable and eco-friendly fuels is accelerating. Propane, recognized as liquefied petroleum gas or LPG, has garnered research interest as an alternative fuel due to its notable advantages, including a high-octane rating, reduced greenhouse gas emissions, and potential cost-effectiveness. However, to realize its full potential as an alternative fuel it is essential to develop catalysts that efficiently handle emissions at low temperatures. In our research, we investigated three distinct palladium (Pd)-based three-way catalyst (TWC) formulations (PdRh, Pd-only, and Pd-OSC) to investigate the influence of typical TWC components rhodium (Rh) and oxygen storage components (OSC) in exhaust scenarios relevant to propane-fueled engines. Among these, the formulation containing oxygen storage components (Pd-OSC) showed the highest reactivity for both NO and C 3 H 8 while minimizing performance degradation from hydrothermal aging (HTA). Notably, the temperature of 50% conversion (T50) for propane in the Pd-OSC fresh and HTA sample was lower by 30 °C and 13 °C, respectively, compared to the Pd-only sample, highlighting the role of oxygen storage materials in enhancing catalyst performance, even without dithering. Additionally, N 2 physisorption showed that the PdOSC sample has a higher surface area and increased pore volume. This underscores the idea that OSC materials not only augment the catalyst’s porosity but also optimize reactant accessibility to active sites, thus elevating catalytic efficiency. In addition to evaluating performance, we further explored the performance and characteristics of the catalysts using catalytic probe reactions, such as water–gas shift and steam reforming reactions.
Reducing embodied carbon emissions in buildings and striving for carbon neutrality through sustainable design has become a primary goal in the construction industry. The industry is expanding its use of wood-engineered products, such as cross-laminated timber (CLT) to achieve those sustainability goals. Furthermore, replacing steel fasteners with lower embodied carbon alternatives is gaining increased attention in timber engineering. This paper aims to quantify the mechanical behavior of single shear-plane CLT-to-CLT joints using hardwood dowels. The study includes 154 single shear-plane experimental tests with four different CLT species, two hardwood dowel species, and two dowel diameters. Further, the test specimens were subjected to cyclic and monotonic loading until failure, and mechanical properties such as yielding and ultimate strength, serviceability and yielding stiffness, and ductility were quantified. The hardwood dowel CLT-to-CLT joints demonstrated high strength with up to 10.3 kN (2.3 kips) capacity per dowel and lateral stiffness of up to 5.2 kN/mm (30.8 k/in) per dowel. This strength and stiffness are roughly equivalent to or exceed values for mass timber wood screws, making hardwood dowels a good alternative. In addition, the observed mean ductility of joints ranged from 2.4 to 4.5. Finally, analytical equations for the joints' yielding strength, lateral serviceability stiffness, and yielding stiffness were derived with non-linear regression analysis and validated using the experimental test results.
The Oil-Free Turbine Engine Technology Project team successfully demonstrated a foil-air bearing designed for the core rotor shaft of a turbine engine. The bearings were subjected to test conditions representative of the engine core environment through a combination of high speeds, sustained loads, and elevated temperatures. The operational test envelope was defined during conceptual design studies completed earlier this year by bearing manufacturer Mohawk Innovative Technologies and the turbine engine company Williams International. The prototype journal foil-air bearings were tested at the NASA Glenn Research Center. Glenn is working with Williams and Mohawk to create a revolution in turbomachinery by developing the world's first Oil-Free turbine aircraft engine. NASA's General Aviation Propulsion project and Williams International recently developed the FJX-2 turbofan engine that is being commercialized as the EJ-22. This core bearing milestone is a first step toward a future version of the EJ-22 that will take advantage of recent advances in foil-air bearings by eliminating the need for oil lubrication systems and rolling element bearings. Oil-Free technology can reduce engine weight by 15 percent and let engines operate at very high speeds, yielding power density improvements of 20 percent, and reducing engine maintenance costs. In addition, with NASA coating technology, engines can operate at temperatures up to 1200 F. Although the project is still a couple of years from a full engine test of the bearings, this milestone shows that the bearing design exceeds the expected environment, thus providing confidence that an Oil-Free turbine aircraft engine will be attained. The Oil-Free Turbomachinery Project is supported through the Aeropropulsion Base Research Program.
Soot emissions in combustion are unwanted consequences of burning hydrocarbon fuels. The presence of soot during and following combustion processes is an indication of incomplete combustion and has several negative consequences including the emission of harmful particulates and increased operational costs. Efforts have been made to reduce soot production in combustion engines through utilizing oxygenated biofuels in lieu of traditional nonoxygenated feedstocks. The ongoing Co-Optimization of Fuels and Engines (Co-Optima) initiative from the US Department of Energy (DOE) is focused on accelerating the introduction of affordable, scalable, and sustainable biofuels and high-efficiency, low-emission vehicle engines. The Co-Optima program has identified a handful of biofuel compounds from a list of thousands of potential candidates. In this study, a shock tube was used to evaluate the performance of soot reduction of five high-performance biofuels downselected by the Co-Optima program. Current experiments were performed at test conditions between 1,700 and 2,100 K and 4 and 4.7 atm using shock tube and ultrafast, time-resolve laser absorption diagnostic techniques. The combination of shock heating and nonintrusive laser detection provides a state-of-the-art test platform for high-temperature soot formation under engine conditions. Soot reduction was found in ethanol, cyclopentanone, and methyl acetate; conversely, an α-diisobutylene and methyl furan produced more soot compared to the baseline over longer test times. For each biofuel, several reaction pathways that lead towards soot production were identified. The data collected in these experiments are valuable information for the future of renewable biofuel development and their applicability in engines.
Bioconversion of lignocellulosic biomass offers a promising alternative to petroleum-based chemical production. However, inefficient xylose utilization and toxic compounds in cellulosic hydrolysate limit microbial fermentation, as the hydrolysate contains substantial amounts of xylose in addition to glucose. To address these challenges, we engineered Issatchenkia orientalis to produce 3-hydroxypropionic acid (3-HP) directly from sorghum hydrolysate under low-pH conditions. A heterologous xylose utilization pathway consisting of XYL1, XYL2, and XYL3 from Scheffersomyces stipitis was introduced into an engineered 3-HP producing strain, enabling efficient conversion of xylose to 3-HP. The engineered strain produced 46.8 g/L 3-HP from sorghum hydrolysate without nutrient supplementation. To eliminate the lag phase under low-pH conditions, fermentation was conducted at pH 6.0 for the first three days, after which pH control was discontinued and in situ 3-HP accumulation buffered the culture. This partial pH control strategy increased 3-HP productivity by 55% from 0.20 to 0.31 g/L∙h, while maintaining low-pH conditions. Introducing an additional copy of XYL2 further increased 3-HP titer to 53.5 g/L and the yield by 33%, from 0.30 to 0.40 g/g sugars, with pH reaching 4.5 at the end of fermentation. This represents one of the highest reported 3-HP titers and yields from cellulosic hydrolysate without additional nutrient supplementation. This work demonstrates a nutrient-independent and low-pH bioprocess for upgrading lignocellulosic hydrolysate into 3-HP, highlighting the industrial potential of engineered xylose-utilizing I. orientalis for sustainable production of platform chemicals from renewable feedstocks.
Biocatalysis has the potential to address the need for more sustainable organic synthesis routes. Pro-tein engineering can tune enzymes to perform in cascade reactions and for efficient synthesis of en-antiomerically enriched compounds, using both natural and new-to-nature reaction pathways. This review highlights recent achievements in biocatal-ysis, especially the development of novel enzymatic syntheses to access versatile small molecule inter-mediates and complex biomolecules. Biocatalytic strategies for the degradation of persistent pollu-tants and approaches for biomass valorization are also discussed. Here, the transition of chemical synthesis to a greener future will be accelerated by imple-menting enzymes and engineering them for high performance and new activities.
Aliphatic amino alcohols such as 6-amino-1-hexanol are potential platform chemicals for a variety of advanced materials, but applications are currently limited by reagent costs. Aliphatic amino alcohols can currently be synthesized from biomass-derived diols at elevated temperatures and pressures using Ru-based catalysts that produce a mixture of amino-alcohol, diamine, and cyclic amine products. Replacing chemical amination with an enzymatic cascade would reduce resource needs and enable reactions under milder conditions. In this work, we characterized a two-enzyme cascade that selectively converts C4–C7 diols to the corresponding amino alcohols under aqueous conditions at room temperature and pressure. By engineering the rate-limiting enzyme and optimizing reaction conditions, we increased amino alcohol production nearly 30-fold, achieving a selectivity of 99%. The same enzyme cascade could also be used to convert amino alcohols into cyclic amines through reduction of the corresponding cyclic imine. This engineered cascade provides a green opportunity to sustainably synthesize asymmetric bifunctional platform chemicals.
Abstract Background Rhodosporidium toruloides is capable of co-utilization of complex carbon sources and robust growth from lignocellulosic hydrolysates. This oleaginous yeast is therefore an attractive host for heterologous production of valuable bioproducts at high titers from low-cost, deconstructed biomass in an economically and environmentally sustainable manner. Here we demonstrate this by engineering R. toruloides to produce the polyketide triacetic acid lactone (TAL) directly from unfiltered hydrolysate deconstructed from biomass with minimal unit process operations. Results Introduction of the 2-pyrone synthase gene into R. toruloides enabled the organism to produce 2.4 g/L TAL from simple media or 2.0 g/L from hydrolysate produced from sorghum biomass. Both of these titers are on par with titers from other better-studied microbial hosts after they had been heavily engineered. We next demonstrate that filtered hydrolysates produced from ensiled sorghum are superior to those derived from dried sorghum for TAL production, likely due to the substantial organic acids produced during ensiling. We also demonstrate that the organic acids found in ensiled biomass can be used for direct synthesis of ionic liquids within the biomass pretreatment process, enabling consolidation of unit operations of in-situ ionic liquid synthesis, pretreatment, saccharification, and fermentation into a one-pot, separations-free process. Finally, we demonstrate this consolidation in a 2 L bioreactor using unfiltered hydrolysate, producing 3.9 g/L TAL. Conclusion Many steps involved in deconstructing biomass into fermentable substrate can be combined into a distinct operation, and directly fed to cultures of engineered R. toruloides cultures for subsequent valorization into gram per liter titers of TAL in a cost-effective manner.