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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 19 records

The Evolution of Energy-Transducing Systems. Studies with an Extremely Halophilic Archaebacterium

The F-type ATPases are found in remarkably similar versions in the energy-transducing membranes of bacteria, chloroplasts and mitochondria (1). Thus, it is likely that they have originated early in the evolution of life, which is consistent with their function as key enzymes of cellular metabolism. The archaea (formerly called archaebacteria) are a group of microorganisms which, as shown by molecular sequencing and biochemical data, have diverged early from the main line of prokaryotic evolution (2). From studies of members of all three major groups of archaea, the halophiles, methanogens and thermoacidophiles, it emerged that they possess a membrane ATPase, which differs from the F-ATPases. The goal of this project was a comparison of the ATPase from the halophilic archaebacterium Halobacterium saccharovorum with the well-characterized F-type ATPases on the molecular level. The results were expected to allow a decision about the nature of archaebacterial ATPases, their classification as one of the known or, alternatively, novel enzyme complex, and possibly a deduction of events during the early evolution of energy-transducing systems.

Stan-Lotter, Helga↗

The evolution of energy-transducing systems. Studies with an extremely halophilic archaebacterium

The F-type ATPases are found in remarkably similar versions in the energy-transducing membranes of eubacteria, chloroplasts, and mitochondria. Thus, it is likely that they have originated early in the evolution of life, which is consistent with their function as key enzymes of cellular metabolism. The archaebacteria are a group of microorganisms which, as shown by molecular sequencing and biochemical data, have diverged early from the main line of prokaryotic evolution. From studies of members of all three major groups of archaebacteria - the halophiles, methanogens, and thermoacidophiles - it emerged that they possess a membrane ATPase which differs from the F-ATPases. The goal of this project was a comparison of the ATPase from the halophilic archaebacterium Halobacterium saccharovorum with the well-characterized F-type ATPases on the molecular level. Amino acid sequences of critical regions of the enzyme were to be determined, as well as immunoreactions of single subunits in the search for common epitopes. The results were expected to allow a decision about the nature of archaebacterial ATPases, their classification as one of the known or, alternatively, novel enzyme complexes, and possibly deduction of events during the early evolution of energy-transducing systems.

Stan-Lotter, Helga↗

The Evolution of Energy-Transducing Systems. Studies with Archaebacteria

The dicyclohexyl carbodiimide (DCCD)- binding site of the membrane ATPase from Halobacterium saccharovorum was investigated during earlier periods of this Cooperative Agreement and was localized to a cyanogen bromide fragment of subunit 2 from amino acids 379 (Glu) to 442 (Met). Although the exact position of the reactive amino acid (probably a glutamic acid) has not yet been determined, the data, together with recently obtained immuno reactions and sequences of Cyanogen Bromide (CNBr) fragments from E.coli F-ATPase, suggested subunit interactions in the halobacterial ATPase which had not been recognized before. They also provided evidence for the presence of a gamma subunit in the halobacterial ATPase, and for a stretch of a amino acids similar to the 'catch' between beta and gamma in bovine F-ATPase. The evolutionary implications of these findings are twofold: first, halobacterial (or archaebacterial) ATPases appear as complex as those from higher organisms - no simpler versions of these membrane enzymes are known to date; second, a monophyletic origin of the energy-transducing ATPases is becoming more apparent, and - together with other data - the split into V- and F-ATPases may have occurred much later than had been previously thought (i.e., after the split into Archaea and Bacteria). Other work included the characterization of an extremely halophilic isolate (Halococcus salifodinae ) from Permian salt sediments. This organism appeared to be an autotrophic halobacterium; its incorporation of C02 was investigated.

Stan-Lotter, Helga↗

The evolution of energy-transducing systems. Studies with an extremely halophilic archaebacterium

The halobacterial ATPase was labeled with C-14-dicyclohexylcarbodiimide and subunit 2 of the enzyme was prepared by electroelution. Subunit 2 was cleaved by several chemical and enzymatic procedures for further preparation of peptides. Immunoreactions (Western blotting) of halobacterial membranes were performed with an antiserum against subunit A of the vacuolar ATPase from Neurospora crassa. A 85 K band (subunit 1) from the membranes of H saccharovorum and from two halobacterial isolates, which were isolated from Permian salt sediments, reacted strongly with the antiserum. The ATPase from the latter isolates resembled the ATPase from H saccharovorum, but had a higher content of acidic amino acids. If it can be verified that the age of the bacterial isolates is in the same range as when deposition of salt occurred, an extremely interesting system for the study of evolutionary questions would be available, since the salt-embedded bacteria presumably did not undergo mutational and selectional events.

Stan-Lotter, Helga↗

The evolution of energy-transducing systems: Studies with archaebacteria

N-ethylmaleimide (NEM) inhibits the ATPase of H. saccharovorum in a nucleotide protectable manner. The bulk of 14C-NEM is incorporated into subunit 1. Inhibition kinetics indicated a single binding site. To determine the sequence around this site, cyanogen bromide peptides of NEM-labeled ATPase enzyme were prepared and separated on Tris-Tricine gels. Autoradiography indicated that the NEM binding site is probably located in a fragment of Mr 10-12 K. This result will be confirmed by N-terminal sequencing of the peptide. Since the cysteinyl residue, to which NEM is bound, may be located at the C-terminal end, purification and proteolytic treatment of the 10 K peptide will be required. One inhibitor of V-type ATPases, fluoresceinisothiocyanate (FITC) inhibited also the ATPase of H. saccharovorum. Preliminary results indicated protection against inhibition by nucleotides. Localization of the binding sited to the major subunits is in progress. An extraction procedure for the membrane sector of the ATPase complex of H. saccharovorum yielded a preparation which was enriched in a peptide of Mr 5 500. Experiments to test the immunological crossreaction with subunit c from the Escherichia coli F-type ATPase and the labeling with 14C-DCCD are currently carried out. Polyclonal antiserum to the smaller of the major subunits of the ATPase from H. saccharovorum (subunit ll) reacts in Western blots strongly with the alpha and beta subunits of the F1 ATPase of E. coli, suggesting highly conserved regions on both types of ATPases. To elucidate further the regions of homology, cyanogen bromide peptides of the beta subunits were prepared for sequence analysis.

Stan-Lotter, Helga↗

The evolution of energy-transducing systems. Studies with archaebacteria

N-ethylmaleimide (NEM) inhibits the ATPase of H. saccharovorum in a nucleotide protectable manner. The bulk of C-14 NEM is incorporated into subunit one. Cyanogen bromide cleavage of labeled subunit one indicated that NEM bound to a peptide of a Mr of about 8,900. Thus, Cys 262 (H. salinarium numbering) may be the NEM binding site. Cyanogen bromide fragments have been submitted for sequencing. To prove the presence of three Cys residues in subunit one, alkaline cleavage following treatment with NTCB was carried out. Thiol reagents such as p-chloro mercuri phenyl sulfonate also inhibited the ATPase. However, this inhibition was not nucleotide-protectable, suggesting a different location and role for the PCMS-sensitive Cys. The proteolipid which was extracted with chloroform/methanol from the membranes of H. saccharovorum cross-reacted with an antiserum against subunit c (the DCCD-binding protein) of Escherichia coli. Following labeling of membranes from H. saccharovorum with C-14 DCCD under conditions, which inhibited ATP synthesis, the isotope was incorporated into one protein of Mr of about 6,500. Thus, the proteolipid of H. saccharovorum and the DCCD-labeled peptide may be identical. If so, these results suggest that the proteolipid is a component of the membrane sector of an archaeal F-type ATP synthase.

Stan-Lotter, Helga↗

Numerical Methods to Evaluate Hyperelastic Transducers: Hexagonal Distributed Embedded Energy Converters

Hexagonal distributed embedded energy converters, also known as hexDEECs, are centimeter-scale energy transducers that leverage variable capacitance to generate electricity when their hyperelastic structure is dynamically deformed. To better understand, characterize, and optimize hexDEEC designs, a series of numerical methods and techniques were developed to model the hyperelastic mechanics of hexDEECs, electrostatic properties, and electricity generation characteristics. The numerical methods developed for the hyperelastic structural analysis were corroborated by empirical results from another study, and the models and equations for capacitance, electrostatic forces, and electrical potential energy were derived from fundamental electrostatic equations. These methods and techniques were implemented within the STAR-CCM+ multiphysics software Version 2020.3 (15.06.008) environment. Results from this analysis revealed methodologies and techniques necessary to model the energy converters, which will enable future exploration and optimization of more specific designs and corresponding applications.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Hybrid piezoelectric energy harvesting transducer system

A hybrid piezoelectric energy harvesting transducer system includes: (a) first and second symmetric, pre-curved piezoelectric elements mounted separately on a frame so that their concave major surfaces are positioned opposite to each other; and (b) a linear piezoelectric element mounted separately on the frame and positioned between the pre-curved piezoelectric elements. The pre-curved piezoelectric elements and the linear piezoelectric element are spaced from one another and communicate with energy harvesting circuitry having contact points on the frame. The hybrid piezoelectric energy harvesting transducer system has a higher electromechanical energy conversion efficiency than any known piezoelectric transducer.

Xu, Tian-Bing↗

General Testing Setup for Hyperelastic Transducers-DEEC-Tec & Marine Renewable Energy: Preprint

Distributed embedded energy conversion technologies (DEEC-Tec), an emerging domain for ocean wave energy conversion technology, is showing promise for a range of applications. Research is being conducted at the National Renewable Energy Laboratory that leverages this domain to investigate the potential of ocean wave energy converters (WECs) constructed from hyperelastic forms of distributable and embeddable energy transducers. These transducers are available in forms such as disks, rectangles, or hexagons and can be combined in various ways to form energy-producing metamaterials and flexible WECs. DEEC-Tec, therefore, could open doors that enhance ocean wave energy conversion in ways not previously thought possible by allowing for many WEC topologies and morphologies. However, the same diversity and adaptability pose challenges for the development of these DEEC-Tec-oriented hyperelastic transducers. A commercial tensile testing setup was not found that was adaptable enough to accommodate the varied transducers while providing precise force control, range of motion, and noncontact data collection. Because of this lack, a comprehensive test rig was designed in house to be used with a 3D laser scanning device-providing contactless measurements while also allowing for different geometries and various uniaxial loadings. This paper and presentation will discuss these unique challenges and the processes for overcoming them to provide a robust and general testing setup for hyperelastic transducers, of any form, for the DEEC-Tec marine renewable energy domain.

DEEC-Tec↗

On archaebacterial ATPase from Halobacterium saccharovorum

The energy transducing ATPase from Halobacterium saccharovorum was studied in order to define the origin of energy transducing systems. The ATPase required high salt concentration (4M NaCl) for activity; activity was rapidly lost when NaCl was below 1 Molar. At low salt concentration, the membrane bound ATPase activity could be stabilized in presence of spermine. However, following solubilization spermine was ineffective. Furthermore, F1 ATPase activity was stabilized by ammonium sulfate even when the NaCl concentration was less than 1 Molar. These studies suggest that stabilization by hydrophobic interactions preceded ionic ones in the evolution of the energy transducing ATPases.

Kristjansson, H.↗

Undulating transducer for energy harnessing and propulsion apparatuses, methods and systems

Embodiments disclosed may serve the function of a pump, a generator, or a propulsion system. Two flexible fins are coupled via their longitudinal inner edges to fin coupling modules. An external fluid flow causes traveling fin undulations. The angle of the fins relative to each other changes as undulations travel along the fins which can be translated through the fin coupling modules into mechanical work, powered by the external fluid flow. The mechanical work of the fin coupling modules may be the cyclical extension and contraction of pistons or bellows incorporated into the fin coupling modules, which may draw ambient fluid during their extension cycles and compress fluid during their contraction cycles to create a pump. The pistons or bellows may also be sealed from ambient fluid but connected through conduits through which contained fluid is exchanged in a continuous circulation of fluid to power a pump or a turbine.

Filardo, Benjamin Pietro↗

Ultrasonic imaging using trapped energy mode Fresnel lens transducers

Trapped-energy focusing transducers operating in the 2-5 MHz range have been fabricated by plating concentric rings of electrodes on a piezoelectric plate. The concentric ring structure acts as a Fresnel lens and can be used to obtain excellent lateral focusing of ultrasonic waves. The trapping is sufficiently strong to permit optimization of electrode spacings to suppress spurious virtual foci and ring sidelobes.

Das, P.↗

Energy transduction in Halobacterium halobium

The properties and functions of the light-energy-transducing purple membrane of Halobacterium halobium are reviewed. Consideration is given to the protein structure and composition of the membrane and the photochemistry of the protein-retinal complex known as bacteriorhodopsin. The role of bacteriorhodpsin in establishing and maintaining an electrochemical (H(+)) gradient is examined, and interactions of this gradient with Na(+) and K(+) gradients, the light-induced transport of amino acids and the light-induced phosphorylation of ADP are considered. Bacteriorhodopsin and the respiratory chain are discussed as alternative sources of energy for the maintenance of the H(+) gradient. Advantages of the Halobacterium purple membrane system for studies of membrane energetics and the confirmation of the chemiosmotic hypothesis are also noted.

Lanyi, J. K.↗

Covalent labeling of the Arabidopsis plasma membrane H + ‐ ATPase reveals 3D conformational changes involving the C‐terminal regulatory domain

The plasma membrane proton pump is the primary energy transducing, electrogenic ion pump of the plasma membrane in plants and fungi. Compared to its fungal counterpart, the plant plasma membrane proton pump's regulatory C‐terminal domain (CTD) contains an additional regulatory segment that links multiple sensory pathways regulating plant cell length through phosphorylation and recruitment of regulatory 14‐3‐3 proteins. However, a complete structural model of a plant proton pump is lacking. Here, we performed covalent labeling with mass spectrometric analysis (CL‐MS) on the Arabidopsis pump AHA2 to identify potential interactions between the CTD and the catalytic domains. Our results suggest that autoinhibition in the plant enzyme is much more structurally complex than in the fungal enzyme.

Blackburn, Matthew R. [Department of Biochemistry ↗

Delayed leaf greening involves a major shift in the expression of cytosolic and mitochondrial ribosomes to plastid ribosomes in the highly phosphorus-use-efficient Hakea prostrata (Proteaceae)

Abstract Background and aims Hakea prostrata (Proteaceae) is a highly phosphorus-use-efficient plant native to southwest Australia. It maintains a high photosynthetic rate at low leaf phosphorus (P) and exhibits delayed leaf greening, a convergent adaptation that increases nutrient-use efficiency. This study aimed to provide broad physiological and gene expression profiles across leaf development, uncovering pathways leading from young leaves as nutrient sinks to mature leaves as low-nutrient, energy-transducing sources. Methods To explore gene expression underlying delayed greening, we analysed a de novo transcriptome for H. prostrata across five stages of leaf development. Photosynthesis and respiration rates, and foliar pigment, P and nitrogen (N) concentrations were determined, including the division of P into five biochemical fractions. Key results Transcripts encoding functions associated with leaf structure generally decreased in abundance across leaf development, concomitant with decreases in foliar concentrations of 85% for anthocyanins, 90% for P and 70% for N. The expression of genes associated with photosynthetic function increased during or after leaf expansion, in parallel with increases in photosynthetic pigments and activity, much later in leaf development than in species that do not have delayed greening. As leaves developed, transcript abundance for cytosolic and mitochondrial ribosomal proteins generally declined, whilst transcripts for chloroplast ribosomal proteins increased. Conclusions There was a much longer temporal separation of leaf cell growth from chloroplast development in H. prostrata than is found in species that lack delayed greening. Transcriptome-guided analysis of leaf development in H. prostrata provided insight into delayed greening as a nutrient-saving strategy in severely phosphorus-impoverished landscapes.

59 BASIC BIOLOGICAL SCIENCES↗

Thiol-based pathways in the thylakoid lumen and their role in photoprotection

The long-term goal of this research is to elaborate the catalysis of thiol-disulfide transactions in the thylakoid lumen, a compartment required for transducing energy via photophosphorylation. The molecular identity of the redox players catalyzing thiol-disulfide exchanges, their relevant targets of action in vivo and how thiol-disulfide chemistry in general controls photosynthesis are the experimental questions this research aims to address. Through previous work, we documented the requirement of catalyzed disulfide formation and reduction for the biogenesis of two photosynthetic enzymes, namely Photosystem II and the cytochrome b6f complex. We established that dedicated trans-thylakoid pathways operate in these processes by delivering reducing and oxidizing power to cysteine-containing subunits of the photosynthetic complexes, which are localized in the lumen compartment. Here we focused on CCDA, CCS4 and CCS5/HCF164, the components of the disulfide-reducing pathway which maintains the heme binding cysteines of apoforms of plastid cytochrome c in the reduced form prior to covalent heme ligation. We showed that CCS4 and CCS5 operate in functionally redundant pathways for the supply of reducing power and postulate a role for CCS4 in recruiting the source of reductants to CCDA or controlling the CCDA-dependent transduction of reducing equivalents. We also demonstrate the disulfide reducing pathway operates to counter thiol oxidation of the heme-binding cysteines by disulfide forming enzyme LTO1 in the thylakoid lumen.

59 BASIC BIOLOGICAL SCIENCES↗

Light energy conservation processes in Halobacterium halobium cells

Proton pumping driven by light or by respiration generates an electrochemical potential difference across the membrane in Halobacterium halobium. The pH changes induced by light or by respiration in cell suspensions are complicated by proton flows associated with the functioning of the cellular energy transducers. A proton-per-ATP ratio of about 3 is calculated from simultaneous measurements of phosphorylation and the proton inflow. This value is compatible with the chemiosmotic coupling hypothesis. The time course of the light-induced changes in membrane potential indicates that light-driven pumping increases a dark pre-existing potential of about 130 mV only by a small amount (20 to 30 mV). The complex kinetic features of the membrane potential changes do not closely follow those of the pH changes, which suggests that flows of ions other than protons are involved. A qualitative model consistent with the available data is presented.

Bogomolni, R. A.↗