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At least 55 records · Page 3

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,

Increasing thermostability of the key photorespiratory enzyme glycerate 3‐kinase by structure‐based recombination

As global temperatures rise, improving crop yields will require enhancing the thermotolerance of crops. One approach for improving thermotolerance is using bioengineering to increase the thermostability of enzymes catalysing essential biological processes. Photorespiration is an essential recycling process in plants that is integral to photosynthesis and crop growth. The enzymes of photorespiration are targets for enhancing plant thermotolerance as this pathway limits carbon fixation at elevated temperatures. We explored the effects of temperature on the activity of the photorespiratory enzyme glycerate kinase (GLYK) from various organisms and the homologue from the thermophilic alga Cyanidioschyzon merolae was more thermotolerant than those from mesophilic plants, including Arabidopsis thaliana. To understand enzyme features underlying the thermotolerance of C. merolae GLYK (CmGLYK), we performed molecular dynamics simulations using AlphaFold-predicted structures, which revealed greater movement of loop regions of mesophilic plant GLYKs at higher temperatures compared to CmGLYK. Based on these simulations, hybrid proteins were produced and analysed. These hybrid enzymes contained loop regions from CmGLYK replacing the most mobile corresponding loops of AtGLYK. Two of these hybrid enzymes had enhanced thermostability, with melting temperatures increased by 6 °C. One hybrid with three grafted loops maintained higher activity at elevated temperatures. Whilst this hybrid enzyme exhibited enhanced thermostability and a similar Km for ATP compared to AtGLYK, its Km for glycerate increased threefold. This study demonstrates that molecular dynamics simulation-guided structure-based recombination offers a promising strategy for enhancing the thermostability of other plant enzymes with possible application to increasing the thermotolerance of plants under warming climates.

59 BASIC BIOLOGICAL SCIENCES

Genetic modification of the shikimate pathway to reduce lignin content in switchgrass ( Panicum virgatum L.) significantly impacts plant microbiomes

Switchgrass (Panicum virgatum L.) is considered a sustainable biofuel feedstock, given its fast-impact growth, low input requirements, and high biomass yields. Improvements in bioenergy conversion efficiency of switchgrass could be made by reducing its lignin content. Engineered switchgrass that expresses a bacterial 3-dehydroshikimate dehydratase (QsuB) has reduced lignin content and improved biomass saccharification due to the rerouting of the shikimate pathway towards the simple aromatic protocatechuate at the expense of lignin biosynthesis. However, the impacts of this QsuB trait on switchgrass microbiome structure and function remain unclear. To address this, wild-type and QsuB-engineered switchgrass were grown in switchgrass field soils, and samples were collected from inflorescences, leaves, roots, rhizospheres, and bulk soils for microbiome analysis. We investigated how QsuB expression influenced switchgrass-associated fungal and bacterial communities using high-throughput Illumina MiSeq amplicon sequencing of ITS and 16S rDNA. Compared to wild-type, QsuB-engineered switchgrass hosted different microbial communities in roots, rhizosphere, and leaves. Specifically, QsuB-engineered plants had a lower relative abundance of arbuscular mycorrhizal fungi (AMF). Additionally, QsuB-engineered plants had fewer Actinobacteriota in root and rhizosphere samples. These findings may indicate that changes in the plant metabolism impact both AMF and Actinobacteriota similarly or potential interactions between AMF and the bacterial community. This study enhances understanding of plant-microbiome interactions by providing baseline microbial data for developing beneficial bioengineering strategies and by assessing nontarget impacts of engineered plant traits on the plant microbiome.

09 BIOMASS FUELS

Kinetic Deep Learning v0.1

Here, we present a method that uses protein levels to predict times series of metabolite concentrations. Understanding this type of pathway dynamics is important in order to predict the behavior of the pathway and, more pragmatically, to be able to design biological systems (such as strains bioengineered to produce chemical products) reliably. Typically, for this purpose, kinetic models consisting of differential equations based on the Michaelis-Menten dynamics have been used in the past. However, these methods can rarely produce good fits to measured data time series. Possibly, this happens because the kinetic constants are unknown or are different from the ones measured in vivo, or perhaps because Michaelis-Menten dynamics is not a satisfactory description. In order to improve the predictive nature of these kinetic models we have eliminated the Michaelis-Menten description of pathway dynamics and we have substituted it by algorithms that automatically learn these dynamics from previously obtained metabolomics and proteomics data using machine learning approaches. Specifically, kinetic deep learning uses deep learning to map proteomics time series to metabolite concentration time series, instead of learning the first metabolite derivative and integrating in (as in the first version of kinetic learning). This approach is shown to provide good to excellent results with a data set specifically collected for this purpose.

Garcia Martin, Hector [Joint BioEnergy Institute (

Development of emerging model microorganisms: Megasphaera elsdenii for biomass and organic acid upgrading to fuels and chemicals

The metabolic diversity of microorganisms in nature represents a largely untapped source of valuable compounds that are difficult or impossible to produce in the limited number of available model systems. Efforts to produce longer-chain alcohols, such as hexanol, in organisms like Escherichia coli have met with limited success; production of C6 and larger products remains low, highlighting the challenges of extending chain elongation pathways beyond a single cycle. Megasphaera elsdenii naturally condenses acetyl-CoA to efficiently generate C4–C8 organic acids, making it a promising candidate for producing fuels and chemicals from lactate and plant-derived carbohydrates. This high native flux through the chain elongation pathway offers the potential for higher yields and titers of medium-chain products, such as hexanol, compared to conventional hosts. Recent advances—most notably the development of a transformation method for M. elsdenii—have further opened the organism to detailed physiological studies and bioengineering. While full development of M. elsdenii as a hexanol-producing platform was not achieved, significant progress was made in understanding its metabolism and building foundational genetic tools for future engineering.

60 APPLIED LIFE SCIENCES

Development of Biofuel and Bioenergy Processes for Photosynthetic CO 2 Assimilation (CRADA Final Report)

In order to fully exploit algae carbon assimilation for fuels, a preqrequisite is high photosynthetic efficiency and carbon conversion to fuel precursors. In the context of this collaborative project between NLR and EMRE we aim to address aspects of both the biological constraints around photosynthesis-driven carbon assimilation flux as well as elements that bridge the gap between laboratory cultivation and meaningful outdoor deployment, building on the unique capabilities of NLR and EMRE’s algae bioengineering teams. The goal of this work is to improve our understanding of key cellular biochemistry, mass cultivation, and biomass conversion constraints that presently limit the potential bio-oil yield. The proposed first phase of this work will focus on biomass growth improvements by testing hypotheses for alleviating identified metabolic and photophysiological bottlenecks. The results of this work could inform decisions by stakeholders (e.g., industry, community, investors) on the design and deployment of a sustainable biofuel supply chain from algae.

09 BIOMASS FUELS

Artificial Intelligence Transforming Post-Translational Modification Research

Post-Translational Modifications (PTMs) are covalent changes to amino acids that occur after protein synthesis, including covalent modifications on side chains and peptide backbones. Many PTMs profoundly impact cellular and molecular functions and structures, and their significance extends to evolutionary studies as well. In light of these implications, we have explored how artificial intelligence (AI) can be utilized in researching PTMs. Initially, rationales for adopting AI and its advantages in understanding the functions of PTMs are discussed. Then, various deep learning architectures and programs, including recent applications of language models, for predicting PTM sites on proteins and the regulatory functions of these PTMs are compared. Finally, our high-throughput PTM-data-generation pipeline, which formats data suitably for AI training and predictions is described. We hope this review illuminates areas where future AI models on PTMs can be improved, thereby contributing to the field of PTM bioengineering.

59 BASIC BIOLOGICAL SCIENCES

Transducer technology transfer to bio-engineering applications

The results of a technology transfer of a miniature unidirectional stress transducer, developed for experimental stress analysis in the aerospace field, to applications in bioengineering are reported. By modification of the basic design and innovations in attachment techniques, the transducer was successfully used in vivo on the myocardium of large dogs to record the change in contractile force due to coronary occlusion, reperfusion, and intervention.

Duran, E. N.

Technical evaluation of the Aerospace Medical Panel Specialists Meeting on Escape Problems and Manoeuvres in Combat Aircraft

A technical evaluation of the papers presented at a conference on escape systems for helicopters and V/STOL aircraft was made. The subjects discussed include the following: (1) bioengineering aspects of spinal injury during ejection, (2) aerodynamic forces acting on crewman during escape, (3) operational practicality of fly away ejection seats, (4) helicopter survivability requirements, (5) ejection experience from V/STOL aircraft, and (6) research projects involving escape and retrieval systems.

Jones, W. L.

Summer Institute in Biomedical Engineering, 1973

Bioengineering of medical equipment is detailed. Equipment described includes: an environmental control system for a surgical suite; surface potential mapping for an electrode system; the use of speech-modulated-white-noise to differentiate hearers and feelers among the profoundly deaf; the design of an automatic weight scale for an isolette; and an internal tibial torsion correction study. Graphs and charts are included with design specifications of this equipment.

Deloatch, E. M.

Rehabilitation and the Veterans' Administration

The Veteran's Administration health care system provides prosthetic and sensory aids for the rehabilitation of neurologically handicapped veterans. Research and development centers include prosthetic clinic teams, orthopedic shops, restoration clinics, bioengineering services, orthotics, etc.

Meister, F.

A perceptual channel for information transfer over kilometer distances Historical perspective and recent research

For more than 100 years, scientists have attempted to determine the truth or falsity of claims for the existence of a perceptual channel whereby certain individuals are able to perceive and describe remote data not presented to any known sense. This paper presents an outline of the history of scientific inquiry into such so-called paranormal perception and surveys the current state of the art in parapsychological research in the United States and abroad. The nature of this perceptual channel is examined in a series of experiments carried out in the Electronics and Bioengineering Laboratory of Stanford Research Institute. The perceptual modality most extensively investigated is the ability of both experienced subjects and inexperienced volunteers to view, by innate mental processes, remote geographical or technical targets including buildings, roads, and laboratory apparatus. The accumulated data indicate that the phenomenon is not a sensitive function of distance, and Faraday cage shielding does not in any apparent way degrade the quality and accuracy of perception. On the basis of this research, some areas of physics are suggested from which a description or explanation of the phenomenon could be forthcoming.

Puthoff, H. E.

Space transportation system flight 2 OSTA-1 scientific payload data management plan

The Shuttle Imaging Radar-A (SIR-A), Shuttle Multispectral Infrared Radiometer (SMIRR), Future Identification and Location Experiment (FILE), Measurement of Air Pollution from Satellites (MAPS), Ocean Color Experiment (OCE), the Night/Day Optical Survey of Lightning (NOSL), and the Heflex Bioengineering Test (HBT) experiments are described.

Source record

OSTA-1 - Shuttle's first scientific and applications payload

The Office of Space and Terrestrial Applications' OSTA-1 scientific and applications payload will be carried by the second Space Shuttle flight and be concerned primarily with the remote sensing of land resources, atmospheric phenomena and ocean conditions. Incorporated by the OSTA-1 payload are the Shuttle Multispectral IR Radiometer, the Feature Identification and Location Experiment, the Measurement of Air Pollution from Satellites experiment, the Ocean Color Experiment, the Night/Day Optical Survey of Lightning, and the Heflex Bioengineering Test.

Demas, L. J.

Life science research in space - The Spacelab era

The events leading up to Spacelab mission 4, which is to be dedicated exclusively to life sciences experimentation in 1984-85, are described. Out of 400 experiment proposals initially received and assessed, 87 candidates were chosen for definition studies to identify the resources which would be required. These proposals addressed such problems encountered in previous space flights as space motion sickness, cardiovascular deconditioning and muscle wasting, calcium loss, and red cell mass reduction. Additional experiments were selected from bioengineering, plant physiology, and cell biology. Human subjects will consist of a Mission Specialist Astronaut and up to four Payload Specialists. Equipment to be used in experimentation includes biotelemetry systems, microscopes, cameras, centrifuges and refrigerators, all of which have been designed for use in weightless conditions. The 87 candidate experiments will be further reduced to about 20.

Farrell, R. M.

The OSTA-1 mission - Overview of the goals, mission parameters and instrumentation for the first Shuttle scientific payload

Aspects of the second Space Shuttle flight, which carried the first payload of scientific instruments for remote sensing of the earth's atmosphere, oceans, and land areas, are discussed. The development and integration of the payload, the mission simulations and end-to-end testing, the launch, the payload mission operations, the landing, data removal, and payload removal, the scientific results, and some planned experiments using the Shuttle are addressed. Summaries of the experimental results are presented, including those for the imaging radar, infrared radiometer, ocean color experiment, air pollution measurements, feature identification and location experiment, night/day optical lightning survey, and bioengineering test. The availability of data and science reports is also covered.

Taranik, J. V.