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At least 19 records

Fuel Property Effects on Stochastic Preignition Events During Engine Load Transitions

Stochastic preignition (SPI) is an abnormal combustion phenomenon that can cause catastrophic engine damage. There have been several proposed mechanisms of SPI, where a uniform source is still not certain, however, SPI tendencies have been shown to be influenced by engine operating conditions, oil composition, engine age, and fuel chemical and physical properties. Laboratory research and testing for SPI propensity is challenging given the stochastic nature of events, as well as the potential for significant degradation of the engine platform and measuring equipment over time. Thus, SPI specific experiments are generally conducted under either sustained or cyclic patterning of steady-state operating conditions to avoid the influence of transient engine boundary conditions on test parameters of interest (e.g. oil additive package, fuel properties, engine speed/load, etc.). In this work a cyclically varying SPI test sequence involves a 5 min engine warmup period at a low engine load of around 4 bar gross indicated mean effective pressure (IMEPg), followed by a transition to high load (~20 bar IMEPg) at a constant 2000 rev/min engine speed for a total of 25 min. This individual test sequence load schedule is then sequentially repeated 10 times to generate significant statistical data for analysis. This work examines the influence of fuel chemical and physical properties on SPI tendency during the unsteady portion of the 10-cycle sequence (the first 5 min of the high load operation in each sequence of the loading cycle) which has been discarded from previous analyses due to the uncertainty in engine operating and thermal boundary conditions. Results from this analysis suggest an increasing trend in the ratio of SPI events during the unsteady test period relative to the steady test period with increasing fuel Reid Vapor Pressure (RVP), implying differences in uncontrolled ignition source terms, possibly from, fuel wall interactions and retention during the load transition phase of the test.

Splitter, Derek [ORNL] (ORCID:0000000174044047)

Rheology and engine performance of very low sulfur fuel oil blended with 10% fast pyrolysis and hydrothermal liquefaction oils in a 2-stroke crosshead engine

The performance and emissions for a downscaled single-cylinder 2-stroke crosshead engine were determined for a very low sulfur fuel oil (VLSFO) when blended with 10 wt.% fast pyrolysis (FP) or hydrothermal liquefaction (HTL) bio-intermediates. The FP and HTL oils were derived from biomass and were observed to contain lower molecular weight (MW) hydrocarbons than neat VLSFO (which was evaluated as a baseline comparison). The addition of either biofuel reduced the overall viscosity of the VLSFO. Aging tests at 50, 90, and 120°C showed that the dynamic viscosity of VLSFO increased with exposure time up to two weeks. Similar trends were observed for the FP and HTL blends, but a pronounced spike in viscosity occurred for these fuels during the early period of exposure. None of the viscosity increases exceeded the operational limits of fuel system pumps. Engine performance studies were conducted under low, medium and high load operational settings. The relative performance of the test fuels was highly dependent on operating condition. In general, the engine results for the three test fuels were similar, but modest improvements in brake thermal efficiency and brake specific fuel consumption were observed, which may be attributed to the heightened reactivity of low molecular weight fraction of the FP and HTL oils.

09 BIOMASS FUELS

Engineered Water for Improved Oil Recovery from Fractured Reservoirs (Final Scientific/Technical Report)

The goal of this proposal is to field test the novel technology of engineered water (modifying ionic composition and/or adding surfactants or nanoparticles) injection to improve oil recovery from the carbonate matrix in fractured reservoirs. Carbonate reservoirs tend to be oil-wet/mixed-wet due to positively charged carbonate mineral surfaces and negatively charged acidic/asphaltic components in the oil. Many carbonate reservoirs are also highly fractured. Water injection does not work efficiently in fractured oil-wet reservoirs because water does not imbibe into the matrix. The water composition can be engineered with ionic modification, surfactants and nanoparticles which can change the wettability of carbonate rocks towards more water-wet conditions which can imbibe water into the matrix and push the oil into the fractures. The engineered water can also lower interfacial tension and minimize capillary forces so that gravity can push the oil up and out of the matrix. Wettability alteration also increases water relative permeability which can also improve oil recovery in non-fractured carbonate reservoirs. This technology, if successful, can be a game changer for West Texas carbonates.

02 PETROLEUM

Single‐Step Recovery of Water from Stable Crude Oil‐Water Emulsion Using Surface Engineered Hybrid Inorganic‐Polymer Membranes

ABSTRACT Separating crude oil from water remains one of the most stubborn challenges in environmental remediation, especially for surfactant‐stabilized emulsions that resist conventional demulsification methods. Here, we report a scalable strategy for achieving near‐zero‐discharge separation of crude oil emulsions using a single superhydrophilic membrane. By applying low‐temperature atomic layer deposition (ALD) of various metal oxides onto activated polyvinylidene fluoride (PVDF) membranes, we create atomically precise surface‐engineered (SE) membranes that maintain an exceptionally strong hydration layer at the membrane‐feed interface, even at high oil loadings. Among the various metal oxides, TiO 2 ‐modified SE membranes exhibit superior interfacial water stability, enabling sustained dewatering of complex crude oil‐water emulsions with >98% separation efficiency and >97% water recovery, compared to only 24.6% water recovery for the pristine membrane. This separation performance surpasses conventional hydrophilic membranes and is comparable to complex Janus channel membrane systems, demonstrating near‐complete emulsion separation using a single membrane. This low‐temperature membrane surface engineering process with atomic‐level precision and potential for scalability via roll‐to‐roll fabrication is promising for industrial‐scale, energy‐efficient water treatment and oil spill remediation applications.

Sengupta, Bratin [Applied Materials Division Argon

Data for Multisite Field Evaluation of Oil Accumulation and Agronomic Performance in Grain and Sweet Sorghums Engineered for Lipid Hyperaccumulation

Oil sorghum (OS) has been developed by engineering grain (TX430) and sweet (Ramada) genetic backgrounds to accumulate triacylglycerols (TAG) in vegetative tissues as an energy-dense feedstock for sustainable aviation fuel (SAF) and other biofuels. This study evaluated two TX430 OS lines (TxHO-2, TxHO-3) and two Ramada OS lines (RmHO-1, RmHO-2) alongside wild-type (WT) lines in NE and IL over 2 years (2023–2024) to quantify genotype × environment effects on agronomic performance and TAG accumulation. Across four environments, TX430 OS lines showed average TAG concentrations of 15.0 g kg−1 in leaves and 12.8 g kg−1 in stems, approximately 19-fold higher than WT. Ramada OS lines accumulated 26.1 g kg−1 in leaves and 12.3 g kg−1 in stems, approximately 25-fold and 13-fold increases over WT, respectively. OS lines in TX430 exhibited an 18% reduction in biomass (8.4 vs. 9.9 Mg ha−1 for WT), while Ramada OS lines had similar WT biomass (18.3 vs. 19.9 Mg ha−1 for WT). Among TX430 OS lines, TxHO-2 achieved the highest TAG yield (190 kg ha−1), while RmHO-1 led the Ramada lines (335 kg ha−1) due to higher biomass and similar TAG concentration. Enhanced TAG accumulation increased N, P, and K removal in TX430 lines but not in Ramada lines. Structural carbohydrate and ash concentration were unaffected. Overall, results confirm vegetative lipid accumulation as a viable strategy for high-biomass sorghum, supporting its potential as a dual-purpose feedstock for SAF. Future work should focus on minimizing biomass yield penalties and improving nutrient use efficiency in oil sorghum systems.

Agronomy

Multisite Field Evaluation of Oil Accumulation and Agronomic Performance in Grain and Sweet Sorghums Engineered for Lipid Hyperaccumulation

Oil sorghum (OS) has been developed by engineering grain (TX430) and sweet (Ramada) genetic backgrounds to accumulate triacylglycerols (TAG) in vegetative tissues as an energy-dense feedstock for sustainable aviation fuel (SAF) and other biofuels. This study evaluated two TX430 OS lines (TxHO-2, TxHO-3) and two Ramada OS lines (RmHO-1, RmHO-2) alongside wild-type (WT) lines in NE and IL over 2 years (2023–2024) to quantify genotype × environment effects on agronomic performance and TAG accumulation. Across four environments, TX430 OS lines showed average TAG concentrations of 15.0 g kg −1 in leaves and 12.8 g kg −1 in stems, approximately 19-fold higher than WT. Ramada OS lines accumulated 26.1 g kg −1 in leaves and 12.3 g kg −1 in stems, approximately 25-fold and 13-fold increases over WT, respectively. OS lines in TX430 exhibited an 18% reduction in biomass (8.4 vs. 9.9 Mg ha −1 for WT), while Ramada OS lines had similar WT biomass (18.3 vs. 19.9 Mg ha −1 for WT). Among TX430 OS lines, TxHO-2 achieved the highest TAG yield (190 kg ha −1 ), while RmHO-1 led the Ramada lines (335 kg ha −1 ) due to higher biomass and similar TAG concentration. Enhanced TAG accumulation increased N, P, and K removal in TX430 lines but not in Ramada lines. Structural carbohydrate and ash concentration were unaffected. Overall, results confirm vegetative lipid accumulation as a viable strategy for high-biomass sorghum, supporting its potential as a dual-purpose feedstock for SAF. Future work should focus on minimizing biomass yield penalties and improving nutrient use efficiency in oil sorghum systems.

60 APPLIED LIFE SCIENCES

Integration of plant and microbial oil processing at oilcane biorefineries for more sustainable biofuel production

Oilcane—an oil-accumulating crop engineered from sugarcane—and microbial oil have the potential to improve renewable oil production and help meet the expected demand for bioderived oleochemicals and fuels. To assess the potential synergies of processing both plant and microbial oils, the economic and environmental implications of integrating microbial oil production at oilcane and sugarcane biorefineries were characterized. Due to decreased crop yields that lead to higher simulated feedstock prices and lower biorefinery capacities, current oilcane prototypes result in higher costs and carbon intensities than microbial oil from sugarcane. To inform oilcane feedstock development, we calculated the required biomass yields (as a function of oil content) for oilcane to achieve financial parity with sugarcane. At 10 dw% oil, oilcane can sustain up to 30% less yield than sugarcane and still be more profitable in all simulated scenarios. Assuming continued improvements in microbial oil production from cane juice, achieving this target results in a minimum biodiesel selling price of 1.34 [0.90, 1.85] USD∙L –1 (presented as median [5th, 95th] percentiles), a carbon intensity of 0.51 [0.47, 0.55] kg CO 2 e L –1 , and a total biodiesel yield of 2140 [1870, 2410] L ha –1 year –1 . Compared to biofuel production from soybean, this outcome is equivalent to 3.0–3.9 as much biofuel per hectare of land and a 57%–63% reduction in carbon intensity. While only 20% of simulated scenarios fell within the market price range of biodiesel (0.45–1.11 USD∙L –1 ), if the oilcane biomass yield would improve to 25.6 DMT∙ha –1 ∙y –1 (an equivalent yield to sugarcane) 87% of evaluated scenarios would have a minimum biodiesel selling price within or below the market price range.

09 BIOMASS FUELS

Persistent Elevated Soot Emissions Induced by Clustered Stochastic Preignition Events

Stochastic Preignition (SPI) is an abnormal combustion phenomenon that can occur in spark-ignition engines particularly under high-load operation. SPI is characterized by uncontrolled initiation of combustion prior to spark discharge, an abnormal combustion process that can lead to severe knock events and significant engine damage. SPI has been associated with fuel properties, lubricant composition, and engine design and operation. Here, in this work, a single-cylinder test engine with a dry-sump oil system was utilized to study the SPI response of E10 and E25 fuels with a range of Reid Vapor Pressure (RVP). An automated test procedure was employed, consisting of ten square-waved load profile segments, with each segment composed of 5 min of low-load operation followed by 25 min of sustained high-load operation. These tests were replicated across multiple days of testing including a lubricant triple flush between tests, and an online Fuel in Oil diagnostic measurement. Exhaust particulate emissions were continuously measured by an AVL microsoot sensor (MSS). Elevated particulate matter emissions were observed to occur concurrently with SPI events as blooms of soot. Particularly after clustered events (i.e., multiple SPI cycles occurring within 10 consecutive engine cycles), high soot emissions were observed to persist over several days of sequential operation despite daily lubricant changes, a complete warm-up procedure, and sustained low-load operation between test segments. This result implies that the particulate emissions trends may be dominated by deposit-based effects, where higher load operation is needed to alter deposition and formation processes. The observed soot blooms were also found to correspond to a reduction in the engine fueling and the fuel engine oil dilution rate despite the engine exhaust remaining at stoichiometric exhaust operation. These observations suggest that post-SPI events, pathways for lubricant migration and consumption into the combustion chamber may occur until these pathways are closed from deposit formation or ring dynamics during extended operation. These observed sooting propensity persisted with all fuels tests, but a linear correlation was observed between the summation of soot and particulate matter index (PMI) value for each fuel as well as SPI events, proving that PMI is a crucial fuel property for reducing SPI.

Splitter, Derek [Oak Ridge National Laboratory (OR

Building on success in systems design of high yielding low‐input energycanes for marginal lands

Energycane and Miscanthus are highly sustainable and productive perennial grasses that produce a large annual crop of shoot biomass. Theoretical analysis of metabolism has suggested that these crops could be engineered to accumulate 20% of their harvestable biomass as oil. Given their much higher biomass yields than soybean and canola, this would yield several times more oil per acre. This could be converted to biodiesel or sustainable aviation fuel (SAF) by well‐established technologies. Energycane is well suited to the southeast quarter of the contiguous 48 states and Miscanthus to the northeast quarter. The objective of this award was to provide the foundation for converting these crops to oilcanes from DNA constructs and transformation to field trials and pilot plant development of oil extraction. Genetic engineering focused on development and implementation of improved constructs to increase oil levels, and increased photosynthesis to support the additional energy needed for oil production. The specific progress in each of our subtasks follows together with the products arising.

09 BIOMASS FUELS

Persistent fatty acid catabolism during plant oil synthesis

Plant lipids are an essential energy source for diets and are a sustainable alternative to petroleum-based fuels and feedstocks. Fatty acid breakdown during seed germination is crucial for seedling establishment but unexpected during seed filling. Here, we demonstrate that the simultaneous biosynthesis and degradation of fatty acids begins early and continues across all phases of oil filling and throughout the photoperiod. Tests in camelina, rapeseed, and an engineered high-oil tobacco line confirmed that concomitant synthesis and breakdown in oil-producing tissues over development is the rule rather than the exception. Furthermore, we show that transgenics, designed to elevate fatty acid biosynthesis, failed to achieve anticipated increases in storage lipid levels due to increased degradation, potentially explaining the underperformance of engineered lines compared to expectations more generally.

coenzyme A

Understanding the dynamic nature of plant lipid anabolic and catabolic metabolism is key to sustainable oilseed engineering

Plant-derived oils are essential sources of reduced carbon and various fatty acid (FA) structures for food, biofuels, and the oleochemical industry. Despite extensive efforts, engineering mainstream oilseed crops to produce high levels of industrially valuable unusual FAs (UFAs) remains challenging. This review synthesizes recent advances in the understanding of lipid metabolic networks, emphasizing how species-specific regulation of FA synthesis, activation, and delivery influences triacylglycerol (TAG) assembly to govern the efficiency of UFA accumulation. Key insights reveal that acyl flux through anabolic and catabolic branches of lipid metabolism is tightly controlled by enzyme substrate selectivities, diacylglycerol (DAG) pool compartmentalization, and metabolic context, including lipid remodeling and degradation pathways. Engineering success is often constrained by incompatibilities between UFA biosynthetic enzymes and endogenous host metabolism, leading to flux imbalances, futile cycles, and undesired phenotypes. We highlight emerging strategies to overcome these barriers, such as the use of UFA-selective acyltransferases, coordinated manipulation of DAG source pools, suppression of competing endogenous enzymes, and exploitation of TAG remodeling mechanisms. This integrated synthesis provides a conceptual framework for logic-based engineering of oilseeds with enhanced UFA content by offering new avenues for sustainable biomanufacturing of valuable lipids.

acyltransferase specificity

Data for Expression of a Bacterial Trehalose 6-Phosphate Synthase Gene otsA in Camelina sativa Seeds Promotes the Channelling of Carbon Towards Oil Accumulation

Improving seed oil yield is essential for developing Camelina sativa as a sustainable biofuel crop. Fatty acid synthesis depends on the production of acetyl-CoA from photosynthetically derived sugars. Trehalose 6-phosphate (T6P), a proxy for sucrose availability, can link sugar status to plant growth and development. Synthesised by trehalose 6-phosphate synthase (TPS) from UDP-glucose and glucose-6-phosphate, T6P plays a regulatory role in metabolism. Our previous studies on Arabidopsis transgenic lines constitutively expressing the E. coli otsA (encoding TPS) showed increased T6P levels and seed triacylglycerol, along with stunted growth. In the present study we express otsA in camelina under the control of a seed-specific Phaseolin promoter. Seeds of the resulting transgenic lines accumulated high levels of T6P, and a 15%–20% increase in total fatty acids and triacylglycerol compared to wild-type. Molecular analysis showed the transgenic seeds had reduced SnRK1 activity, elevated WRI1 protein levels, and increased the levels of WRI1 and its target genes, along with enhanced rates of fatty acid synthesis that increased seed weights relative to wild type. Notably, the increase in oil did not affect seed protein levels but did reduce the soluble metabolite fraction. Crucially, seed-specific expression of otsA mitigated the growth defects associated with constitutive otsA expression, and the transgenic lines showed normal seed development and germination. These findings demonstrate that targeted T6P modulation via seed-specific otsA expression is an effective metabolic engineering strategy to boost oil production in camelina and potentially in other oilseed crops and bioenergy crops such as energycane, sorghum and miscanthus.

Lipids

Complete replacement of Arabidopsis oil-producing enzymes with heterologous diacylglycerol acyltransferases

Acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1) and phospholipid:diacylglycerol acyltransferase 1 (PDAT1) share responsibility for triacylglycerol (TAG) biosynthesis, and their selectivities control TAG fatty acid (FA) compositions. For rational metabolic engineering of seed oils, replacing endogenous TAG biosynthesis with exogenous enzymes containing different substrate FA selectivities is desirable; however, the dgat1-1/pdat1-2 double mutant is pollen lethal. Here, we evaluated the ability of 3 DGAT1s, from phylogenetically diverse plants with distinct TAG assembly processes, to completely replace endogenous TAG biosynthesis in Arabidopsis ( Arabidopsis thaliana ). We transformed dgat1-1 mutant plants with expression constructs for DGAT1 s from Camelina sativa , Physaria fendleri , and castor ( Ricinus communis ). Transgene expression was properly “contextualized” by using a previously determined minimum necessary expression unit containing the promoter/5′ UTR and first intron of native AtDGAT1 ; both of these DNA elements are essential for pollen expression. Next, we crossed homozygous lines with a DGAT1/DGAT1/PDAT1/pdat1-2 parent. C. sativa and P. fendleri DGAT1s restored the FA compositions and transcriptional differences of dgat1-1 to near wild-type and rescued the dgat1-1/pdat1-2 pollen lethality. R. communis DGAT1 was active in dgat1-1 seeds but produced unique oil profiles and alterations in the expression of lipid metabolic genes; it also failed to rescue dgat1-1/pdat1-2 lethality. This study confirms that the promoter and first intron of AtDGAT1 can modulate the expression of foreign DGAT1 genes to fit the correct spatiotemporal profile necessary for completely replacing endogenous TAG biosynthesis. Furthermore, it demonstrates an additional layer of unexpected enzyme incompatibility between oilseed lineages, which may complicate bioengineering approaches that seek to replace essential genes with orthologs.

McGuire, Sean T. [Washington State Univ., Pullman,

Data for Development of Vegetative Oil Sorghum: From Lab-to-Field

Biomass crops engineered to accumulate energy-dense triacylglycerols (TAG or ‘vegetable oils’) in their vegetative tissues have emerged as potential feedstocks to meet the growing demand for renewable diesel and sustainable aviation fuel (SAF). Unlike oil palm and oilseed crops, the current commercial sources of TAG, vegetative tissues, such as leaves and stems, only transiently accumulate TAG. In this report, we used grain (Texas430 or TX430) and sugar-accumulating ‘sweet’ (Ramada) genotypes of sorghum, a high-yielding, environmentally resilient biomass crop, to accumulate TAG in leaves and stems. We initially tested several gene combinations for a ‘push-pull-protect’ strategy. The top TAG-yielding constructs contained five oil transgenes for a sorghum WRINKLED1 transcription factor (‘push’), a Cuphea viscosissima diacylglycerol acyltransferase (DGAT; ‘pull’), a modified sesame oleosin (‘protect’) and two combinations of specialized Cuphea lysophosphatidic acid acyltransferases and medium-chain acyl-acyl carrier protein thioesterases. Though intended to generate oils with medium-chain fatty acids, engineered lines accumulated oleic acid-rich oil to amounts of up to 2.5% DW in leaves and 2.0% DW in stems in the greenhouse, 36-fold and 49-fold increases relative to wild-type (WT) plants, respectively. Under field conditions, the top-performing event accumulated TAG to amount to 5.5% DW in leaves and 3.5% DW in stems, 78-fold and 58-fold increases, respectively, relative to WT TX430. Transcriptomic and fluxomic analyses revealed potential bottlenecks for increased TAG accumulation. Overall, our studies highlight the utility of a lab-to-field pipeline coupled with systems biology studies to deliver high vegetative oil sorghum for SAF and renewable diesel production.

Biofuels

Development of vegetative oil sorghum: From lab‐to‐field

Summary Biomass crops engineered to accumulate energy‐dense triacylglycerols (TAG or ‘vegetable oils’) in their vegetative tissues have emerged as potential feedstocks to meet the growing demand for renewable diesel and sustainable aviation fuel (SAF). Unlike oil palm and oilseed crops, the current commercial sources of TAG, vegetative tissues, such as leaves and stems, only transiently accumulate TAG. In this report, we used grain (Texas430 or TX430) and sugar‐accumulating ‘sweet’ (Ramada) genotypes of sorghum, a high‐yielding, environmentally resilient biomass crop, to accumulate TAG in leaves and stems. We initially tested several gene combinations for a ‘push‐pull‐protect’ strategy. The top TAG‐yielding constructs contained five oil transgenes for a sorghum WRINKLED1 transcription factor (‘push’), a Cuphea viscosissima diacylglycerol acyltransferase (DGAT; ‘pull’), a modified sesame oleosin (‘protect’) and two combinations of specialized Cuphea lysophosphatidic acid acyltransferases and medium‐chain acyl‐acyl carrier protein thioesterases. Though intended to generate oils with medium‐chain fatty acids, engineered lines accumulated oleic acid‐rich oil to amounts of up to 2.5% DW in leaves and 2.0% DW in stems in the greenhouse, 36‐fold and 49‐fold increases relative to wild‐type (WT) plants, respectively. Under field conditions, the top‐performing event accumulated TAG to amount to 5.5% DW in leaves and 3.5% DW in stems, 78‐fold and 58‐fold increases, respectively, relative to WT TX430. Transcriptomic and fluxomic analyses revealed potential bottlenecks for increased TAG accumulation. Overall, our studies highlight the utility of a lab‐to‐field pipeline coupled with systems biology studies to deliver high vegetative oil sorghum for SAF and renewable diesel production.

59 BASIC BIOLOGICAL SCIENCES

Biological Design of Lemnaceae aquatic plants for biodiesel production (Final Technical Report)

Support from this award enabled us to expand critical genomic and epigenomic resources available for the study and engineering of duckweeds, and to complete design-build-test-learn cycles that culminated in the engineering of high oil duckweeds. We shared our findings with the public and scientific community in a series of peer-reviewed publications highlighted within. We successfully reengineered metabolism, dramatically increasing oil content by diverting a substantial portion of accumulated carbon from starch. This sets the stage for the use of fast growing aquatic duckweeds as robust, highly productive sources of biodiesel, with the capacity to outperform soybean oil crops.

09 BIOMASS FUELS

Novel Modular Heat Engines with sCO 2 Bottoming Cycle Utilizing Advanced Oil-Free Turbomachinery

This report presents the thermal, mechanical, and electric machine design of a 27krpm rotating test rig aimed at reducing technical risks with hermetic oil-free super-critical carbon dioxide (sCO 2 ) turbomachinery. The test rig rotor system is based on a conceptual turbomachine design discussed that evaluates a sCO 2 waste heat recovery (WHR) unit for land-based gas turbines at natural gas (NG) compressor stations. The key novelty of the hermetic sCO 2 turbomachine concept is the utilization of additively manufactured CO 2 gas bearings and a CO 2 -immersed direct-drive permanent magnet (PM) electric machine. The previous effort on conceptual design identified 3 main technical risks, which included thrust bearing load capacity, rotordynamics, and thermal performance of the system. Therefore, key requirements of the test vehicle include the ability to test the rotor system in a 400psi (27.6 bar) hermetic CO 2 operating environment, apply up to 1,500 lbs (6.7 kN) of rotor thrust loads, capability to assess rotordynamics with radial gas bearings, inclusion of instrumentation to assess thrust bearing load capability, and flexibility for varying secondary cooling flows to confirm thermal model predictions. Design topics such as rotordynamics, bearing design, and electric machine design are addressed, while highlighting critical elements. In addition, additive build trials for Inconel 718 radial and thrust bearings were performed to prove the manufacturability of the bearing design concepts. The results of the design efforts yield a test rig concept with ability to operate at 27,000 rpm, apply thrust loads to 1,500 lbs, deliver CO 2 up to 800psi, and modulate cooling flows.

03 NATURAL GAS

Expression of a Bacterial Trehalose 6-Phosphate Synthase Gene otsA in Camelina sativa Seeds Promotes the Channelling of Carbon Towards Oil Accumulation

Improving seed oil yield is essential for developing Camelina sativa as a sustainable biofuel crop. Fatty acid synthesis depends on the production of acetyl-CoA from photosynthetically derived sugars. Trehalose 6-phosphate (T6P), a proxy for sucrose availability, can link sugar status to plant growth and development. Synthesised by trehalose 6-phosphate synthase (TPS) from UDP-glucose and glucose-6-phosphate, T6P plays a regulatory role in metabolism. Our previous studies on Arabidopsis transgenic lines constitutively expressing the E. coli otsA (encoding TPS) showed increased T6P levels and seed triacylglycerol, along with stunted growth. In the present study we express otsA in camelina under the control of a seed-specific Phaseolin promoter. Seeds of the resulting transgenic lines accumulated high levels of T6P, and a 15%–20% increase in total fatty acids and triacylglycerol compared to wild-type. Molecular analysis showed the transgenic seeds had reduced SnRK1 activity, elevated WRI1 protein levels, and increased the levels of WRI1 and its target genes, along with enhanced rates of fatty acid synthesis that increased seed weights relative to wild type. Notably, the increase in oil did not affect seed protein levels but did reduce the soluble metabolite fraction. Crucially, seed-specific expression of otsA mitigated the growth defects associated with constitutive otsA expression, and the transgenic lines showed normal seed development and germination. These findings demonstrate that targeted T6P modulation via seed-specific otsA expression is an effective metabolic engineering strategy to boost oil production in camelina and potentially in other oilseed crops and bioenergy crops such as energycane, sorghum and miscanthus.

59 BASIC BIOLOGICAL SCIENCES