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At least 55 records · Page 3

The purification and properties of a cd-cytochrome nitrite reductase from Paracoccus halodenitrificans

Paracoccus halodenitrificans, grown anaerobically in the presence of nitrite, contained membrane and cytoplasmic nitrite reductases. When assayed in the presence of phenazine methosulfate and ascorbate, the membrane-bound enzyme produced nitrous oxide whereas the cytoplasmic enzyme produced nitric oxide. When both enzymes were assayed in the presence of methyl viologen and dithionite, the cytoplasmic enzyme produced ammonia. Following solubilization, the membrane-bound enzyme behaved like the cytoplasmic enzyme, producing nitric oxide in the presence of phenazine methosulfate and ascorbate, and ammonia when assayed in the presence of methyl viologen and dithionite. The cytoplasmic and membrane-bound enzymes were purified to essentially the same specific activity. Only a single nitrite-reductase activity was detected on electrophoretic gels and the electrophoretic behavior of both enzymes suggested they were identical. The spectral properties of both enzymes suggested they were cd-type cytochromes. These data suggest that the products of nitrite reduction by the cd-cytochrome nitrite reductase are determined by the location of the enzyme and the redox potential of the electron donor.

NASA Center ARC↗

Identification of a small tetraheme cytochrome c and a flavocytochrome c as two of the principal soluble cytochromes c in Shewanella oneidensis strain MR1

Two abundant, low-redox-potential cytochromes c were purified from the facultative anaerobe Shewanella oneidensis strain MR1 grown anaerobically with fumarate. The small cytochrome was completely sequenced, and the genes coding for both proteins were cloned and sequenced. The small cytochrome c contains 91 residues and four heme binding sites. It is most similar to the cytochromes c from Shewanella frigidimarina (formerly Shewanella putrefaciens) NCIMB400 and the unclassified bacterial strain H1R (64 and 55% identity, respectively). The amount of the small tetraheme cytochrome is regulated by anaerobiosis, but not by fumarate. The larger of the two low-potential cytochromes contains tetraheme and flavin domains and is regulated by anaerobiosis and by fumarate and thus most nearly corresponds to the flavocytochrome c-fumarate reductase previously characterized from S. frigidimarina to which it is 59% identical. However, the genetic context of the cytochrome genes is not the same for the two Shewanella species, and they are not located in multicistronic operons. The small cytochrome c and the cytochrome domain of the flavocytochrome c are also homologous, showing 34% identity. Structural comparison shows that the Shewanella tetraheme cytochromes are not related to the Desulfovibrio cytochromes c(3) but define a new folding motif for small multiheme cytochromes c.

Cytochrome c Group/chemistry/genetics/metabolism↗

Isolation, characterization, and primary structure of rubredoxin from the photosynthetic bacterium, Heliobacillus mobilis

Rubredoxin is a small nonheme iron protein that serves as an electron carrier in bacterial systems. Rubredoxin has now been isolated and characterized from the strictly anaerobic phototroph, Heliobacillus mobilis. THe molecular mass (5671.3 Da from the amino acid sequence) was confirmed and partial formylation of the N-terminal methionyl residue was established by matrix-assisted laser desorption mass spectroscopy. The complete 52-amino-acid sequence was determined by a combination of N-terminal sequencing by Edman degradation and C-terminal sequencing by a novel method using carboxypeptidase treatment in conjunction with amino acid analysis and laser desorption time of flight mass spectrometry. The molar absorption coefficient of Hc. mobilis rubredoxin at 490 nm is 6.9 mM-1 cm-1 and the midpoint redox potential at pH 8.0 is -46 mV. The EPR spectrum of the oxidized form shows resonances at g = 9.66 and 4.30 due to a high-spin ferric iron. The amino acid sequence is homologous to those of rubredoxins from other species, in particular, the gram-positive bacteria, and the phototrophic green sulfur bacteria, and the evolutionary implications of this are discussed.

NASA Discipline Number 52-30↗

Isolation, characterization, and amino acid sequences of auracyanins, blue copper proteins from the green photosynthetic bacterium Chloroflexus aurantiacus

Three small blue copper proteins designated auracyanin A, auracyanin B-1, and auracyanin B-2 have been isolated from the thermophilic green gliding photosynthetic bacterium Chloroflexus aurantiacus. All three auracyanins are peripheral membrane proteins. Auracyanin A was described previously (Trost, J. T., McManus, J. D., Freeman, J. C., Ramakrishna, B. L., and Blankenship, R. E. (1988) Biochemistry 27, 7858-7863) and is not glycosylated. The two B forms are glycoproteins and have almost identical properties to each other, but are distinct from the A form. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis apparent monomer molecular masses are 14 (A), 18 (B-2), and 22 (B-1) kDa. The amino acid sequences of the B forms are presented. All three proteins have similar absorbance, circular dichroism, and resonance Raman spectra, but the electron spin resonance signals are quite different. Laser flash photolysis kinetic analysis of the reactions of the three forms of auracyanin with lumiflavin and flavin mononucleotide semiquinones indicates that the site of electron transfer is negatively charged and has an accessibility similar to that found in other blue copper proteins. Copper analysis indicates that all three proteins contain 1 mol of copper per mol of protein. All three auracyanins exhibit a midpoint redox potential of +240 mV. Light-induced absorbance changes and electron spin resonance signals suggest that auracyanin A may play a role in photosynthetic electron transfer. Kinetic data indicate that all three proteins can donate electrons to cytochrome c-554, the electron donor to the photosynthetic reaction center.

NASA Discipline Exobiology↗

Synthesis and Near IR Photoluminescence of Os(II) Bis(2,2'-bipyridine) (3,8-Diarylethynyl-1,10-phemanthroline) Complexes: Anomalous Behavior of the 3,8-Dinitrophenylethynyl-substituted Homologue

A large bathochromic shift (50 nm) and emission in the near infrared is observed by attaching arylethynyl groups at the 3,8-positions of the 1,10-phenanthroline ligand (phen) of [Os(bipy)2(phen)]2+ (where bipy = 2,2'-bipyridine). Thus [Os(bipy)2(3,8-di-4-methoxyphenylethynyl-1,10-phenathroline)]2+ emits at 795 nm, while [Os(bipy)2(3,8-diphenylethynyl-1,10-phenanthroline)]2+ emits at 815 nm. According to this trend it would have been expected that [Os(bipy)2(3,8-di-4-nitrophenylethynyl-1,10-phenathroline)]2+ emits farther in the near infrared. Nevertheless, this complex is not photoluminescent because of intramolecular electron transfer quenching of the MLCT excited state by the nitroaromatic group. These results set structural and redox potential standards in the design of near infrared emitters based on [Os(bipy)2(phen)]2+ type complexes.

Yang, Jinhua↗

Correlation of Spectral and Electrochemical Properties of a Series of Ferrocene Derivatives

Charge-transfer-to-solvent (CTTS) bands are observed in the UV/VIS spectra of a series of ferrocene derivatives (ferrocene; 1,1’-dimethyl; 1,2,3,4,5-pentamethyl; 1,2,3,4,l’,2’,3’,4’-octamethyl; and decamethyl) by the addition of increasing amounts of CCl4 to ethanol solutions. A linear correlation (slope = 8540 per cm/V) was found between the redox potential and the energy of the CTTS band, consistent with electrochemical and photochemical oxidation by removal of an electron from the same molecular orbital. Inclusion of literature data for ruthenocene and [(η^(5)-C5H5)Fe(CO)]4 results in a line with a slope of 8140 per cm/V, within 1 percent of the wavenumber to electron volt conversion factor. Calculation of association constants (K) shows a slight decrease (1.2 to 0.7 per mole fraction) as the bulkiness of the cyclopentadienyl ring increases, consistent with either a steric or a repulsive electronic effect. The extinction coefficient of the CTTS absorption was constant at approximately 1700 per Mcm .

ultraviolet spectroscopy↗

Enabling Experimental Evolution: Multi-Parameter Sensor System Integration into a Culture/Stressor Biofluidics System

Experimental evolution (EE) exposes microbial communities to ecological stressors, simulating dynamics up to near-extinction events. Combined with comparative sequencing and other molecular tools, such data can inform the genetic and other biological mechanisms underlying extremophile adaptation, and other observed effects. Automating this type of experiment using biofluidics can mitigate many traditional obstacles, including delays in assay results and environment adjustment and the need for many replicates. A first-generation device for automating EE procedures, the Automated Adaptive Directed Evolution Chamber (AADEC), was developed at NASA Ames. UV-C radiation was the stressor, an LED-photodiode array measured optical density, magnetic agitation and peristaltic pump systems ensured nutrient availability, and Arduino microcontrollers provided control. Escherichia coli in LB kanamycin media was used for testing and performance verification. A manual laboratory procedure with timed exposure to UV-C was performed to typify tolerance acquisition. Approximately a 106 factor increase in survival ratio was recorded over multiple iterations. Currently, a second-generation device is being developed integrating more real-time sensors: redox potential (ORP), indicating available/consumed metabolic energy; dissolved oxygen (DO), indicating aerobic/anaerobic growth; pH, indicating metabolic products; and electrical conductivity (EC), another indicator of metabolic products. The EC sensor system was constructed and calibrated in-house and matched commercial sensors in the required range. A Raspberry Pi computer automated the electrical system, allowing real-time data acquisition. The fluidics card was made of CNC-milled polycarbonate for biocompatibility. Each sensor parameter can also be used as a selection pressure alone or in combination with others to create extreme microbial environments. As a proof of concept, this work demonstrated sensor operation in one pair of growth-sensor chambers. It can be expanded to a multi-chamber system to enable inter-culture comparisons and multi-population studies. The prior Arduino system will be ported to the RPi system. Future stressors to be added include thermal, reactive oxygen species, and varying nutrient availability.

Govinda Raj, Chinmayee↗

Investigating Molecular Responses to Space Radiation for Biological Missions Beyond Low Earth Orbit

As we plan crewed missions to the Moon, Mars, and beyond, it is essential to understand how persistent exposure to deep space radiation affects biology. Unlike on the International Space Station (ISS), where crew support and sample return are possible, experiments for long-duration missions require autonomous systems with no sample return. Human cells would be ideal biosensors, but limitations in culture methods, extended prelaunch storage, and long flight durations make it difficult to keep human cells alive. Unlike other model systems, yeast can survive the constraints of long-duration spaceflight. Despite a billion years of evolution separating yeast from humans, we share homology in hundreds of genes important for basic cell function, including responses to DNA damage. Thus, yeast are excellent biosensors for detecting types/extent of damage induced by space radiation. BioSentinel is NASA’s latest biological CubeSat, and first interplanetary space bioscience mission. BioSentinel is launching on Artemis 1, the first flight of NASA’s Space Launch System, in 2022. The BioSensor payload within BioSentinel contains two yeast strains. The wild type serves as a control for health and normal DNA damage repair (DDR). The rad51 deletion mutant is defective for DDR and will undergo alterations to growth and metabolism as it accumulates radiation damage. Changes in growth and metabolic activity will be measured using a 3-color LED detection system and the metabolic redox dye alamarBlue®. Preliminary tests indicate a significant change in alamarBlue responses to space-like, low-dose ionizing radiation. We will discuss these findings in five parts – Introduction to NASA’s biological CubeSats and BioSentinel (presented by Sergio Santa Maria), analysis of flight data from the ISS mission (presented by Kylie Akiyama), preliminary molecular responses to space radiation (presented here), a deeper dive into those pathways (presented by Kyra Keenan), and characterizing stress response through redox potential data (presented by Diana Gentry).

Lauren Courtney Liddell↗

Constraining the Hydration of X-Ray Amorphous and Clay Mineral Phases in Gale Crater, Mars

Clay minerals and salts in the Gale crater stratigraphy investigated by the Mars Science Laboratory (MSL) Curiosity rover preserve information about sediment sources, depositional environment, and post-depositional alteration history. The compositions of amorphous components in samples have bearing on the redox potential, pH, and chemistry of alteration fluids, and on the heterogeneity of these conditions. Interlayer clay hydration in present-day Gale crater could be an indication of high water/rock ratios of late-stage fluid alteration events, which increases the potential habitability of these post-depositional environments. The MSL Dynamic Albedo of Neutrons instrument (DAN) measures bulk hydration and here we attempt to determine the hydration states of both clay minerals and amorphous components throughout the Curiosity traverse to constrain the abundance of individual amorphous phases and determine if clay interlayer water is present today. These implications are also more broadly applicable to similar sedimentary successions outside Gale crater. In this study, we compare hydration results from DAN to mineralogical results from the MSL Chemistry and Mineralogy instrument (CheMin) for 16 different fluvio-lacustrine sample locations. These samples typically contain one or more hydrated crystalline phases, a hydrated X-ray amorphous fraction composed of multiple amorphous phases, and one or more clay mineral phases which CheMin has consistently measured as collapsed (i.e., no interlayer water). However, Gale samples are known to dehydrate during sample acquisition and handling, and because DAN measures hydration to a depth of tens of cm, we can test for the presence of clay interlayer water in situ by comparing DAN bulk hydration to CheMin mineralogy. We can also constrain the hydration of the samples’ amorphous fractions, and with geochemical data from the ChemCam and APXS instruments, this will allow us to better define the phases which compose this fraction.

S Czarnecki↗

A Multi-Sensor, Low Volume, Automated Culture System for Space Biology Experiments

Optical density (absorbance) is commonly used in microbiology to measure cell concentration and other colorimetric indicators. It is the main type of bioscience data returned from the BioSentinel small spacecraft mission, which was recently launched as a secondary payload on Artemis I. The flight unit takes absorbance measurements of yeast cultures at 570 nm, 630 nm, and 850 nm to track cell concentration and oxidation-reduction (redox) state of the metabolic indicator dye alamarBlue over time [1]. Because the data returned from a small spacecraft mission is necessarily limited, BioSentinel uses a specialized ground-based experiment apparatus to map the absorbance data onto several other parameters. This culture apparatus is composed of a polycarbonate base and lid which contain ports for several commercial sensors, including a standard 12 mm dissolved oxygen probe, gaseous CO2, fluid pressure, and two micro-probes for measuring pH and redox potential. It also contains a pressure relief valve, a rubber membrane for culture interaction, and a 12mm port for the custom optical probe designed to match in-flight measurements. Modules in two corners expand the culture volume into narrow columns, enabling solid-state optical density measurement and dielectric spectroscopy. The apparatus can be fit to a plate for magnetic stirring. The apparatus’s optical probe allows mapping between the flight unit’s optical data and the additional parameters measured by the ground-based experiments. Within the probe, a linear actuator pulls a plunger, drawing culture volume into the optical path. The actuator then pushes out the plunger and culture volume, cleaning any stray yeast from the inside surface of the probe. The probe chassis is 3D-printed from biocompatible resin with the optoelectronics cured inside an optically clear conformal coat which functions as a lens. To optimize the probe’s dimensions, including optical path, a test harness was developed to manipulate and measure distance while the LED light sources and photodiode detector are in operation. The test harness was composed of two 3/64” thick polycarbonate plates mounted to a set of calipers. The LEDs and photodiode were mounted on the outside of each plate. A cuvette of sample fluid was then clamped between the plates in line between the LEDs and photodiode. The test harness generated a 78% signal change between water and a dye standard for the green (570 nm) LED, a 90% signal change for the red (630 nm) LED, and a 25% signal change between water and overgrown yeast for the infrared (850 nm) LED. The improvements to the ground experiment apparatus (culture volume reduction, inclusion of an optical probe to measure flight-like optical data, and magnetic stirring for culture homogeneity) allow for more flight-like measurements to be taken and more accurate mapping of these additional parameters to the optical data returned from the flight unit. This enhancement to mission science return will give insight into how deep space radiation may affect human biology for future long-term space exploration.

Multi-Sensor↗

Evolved Gas Analyses of Sedimentary Rocks from the Glen Torridon Clay-Bearing Unit, Gale Crater, Mars: Results From the Mars Science Laboratory Sample Analysis at Mars Instrument Suite

Evolved gas analysis (EGA) data from the Sample Analysis at Mars (SAM) instrument suite indicated Fe-rich smectite, carbonate, oxidized organics, Fe/Mg sulfate, and chloride in sedimentary rocks from the Glen Torridon (GT) region of Gale crater that displayed phyllosilicate spectral signatures from orbit. SAM evolved H2O data indicated that the primary phyllosilicate in all GT samples was an Fe-rich dioctahedral smectite (e.g., nontronite) with lesser amounts of a phyllosilicate such as mixed layer talc-serpentine or greenalite-minnesotaite. CO(2) data supported the identification of siderite in several samples, and CO(2) and CO data was also consistent with trace oxidized organic compounds such as oxalate salts. SO(2) data indicated trace and/or amorphous Fe sulfates in all samples and one sample may contain Fe sulfides. SO(2) data points to significant Mg sulfates in two samples, and lesser amounts in several other samples. A lack of evolved O(2) indicated the absence of oxychlorine salts and Mn3+/Mn4+ oxides. The lack of, or very minor, evolved NO revealed absent or very trace nitrate/nitrite salts. HCl data suggested chloride salts in GT samples. Constraints from EGA data on mineralogy and chemistry indicated that the environmental history of GT involved alteration with fluids of variable redox potential, chemistry and pH under a range of fluid-to-rock ratio conditions. Several of the fluid episodes could have provided habitable environmental conditions and carbon would have been available to any past microbes though the lack of significant N could have been a limiting factor for microbial habitability in the GT region.

A. C. McAdam↗

Metabolic Vessel for Impedance Spectroscopy and Electrochemistry (MVISE): The Ground Mapping Unit for the Lunar Explorer Instrument for Space Biology Applications (LEIA)​

The BioSensor payload on the upcoming LEIA platform aboard a CLPS lander will carry yeast to the moon to study response to radiation and lunar gravity. The LEIA BioSensor is designed to monitor metabolic activity using absorbance in conjunction with alamarBlue for measuring colorimetric changes as proxy measurement for redox potential. The science data returned from small spacecraft mission modules like LEIA is limited as it relies solely on optical measurements, necessitating a corresponding ground mapping unit that is equipped with multiple electrochemical sensors for accurate mapping of the optical data and operates fully automatically. This technology development work discusses the extensive design and optimization efforts put into the ground mapping unit, MVISE. MVISE is a custom designed, 3D-printed vessel with an agitation system, with six different electrochemical sensor probes, and ports for sample collection and a pressure release valve. The sensors provide real-time data, with dry absorbance measurements aligning with LEIA flight hardware and wet measurements demonstrating invasive sensor design enabling a comparison between the two setups. The 3D printer resin was tested for mechanical robustness, biocompatibility, and resistance to autoclave sterilization. The inner walls were coated with food-grade epoxy, ensuring a smooth finish to prevent microbial lodging and dye staining. The MVISE has successfully passed a week-long leak test and is now undergoing active biology tests. The MVISE prototype will be prepared for radiation tests, with three identical units being tested for varying radiation levels and culture compositions at the NASA Space Radiation Laboratory in November 2024. The integrated sensor approach proposed in this work will enable the accurate mapping of the BioSentinel/LEIA optical flight data to six sensor parameters on the ground unit for better science data return and will enable the first effort to evaluate classical biochemical sensor measurements by comparing and contrasting their responses.

Chinmayee Govinda Raj↗

Redox-Active Star Molecules Incorporating the 4-Benzoylpyridinium Cation - Implications for the Charge Transfer Along Branches vs. Across the Perimeter in Dendrimer

Dendrimers are self-repeating globular branched star molecules, whose fractal structure continues to fascinate, challenge, and inspire. Functional dendrimers may incorporate redox centers, and potential applications include antennae molecules for light harvesting, sensors, mediators, and artificial biomolecules. We report the synthesis and redox properties of four star systems incorporating the 4-benzoyl-N-alkylpyridinium cation; the redox potential varies along the branches but remains constant at fixed radii. Bulk electrolysis shows that at a semi-infinite time scale all redox centers are electrochemically accessible. However, voltammetric analysis (cyclic voltammetry and differential pulse voltammetry) shows that on1y two of the three redox-active centers in the perimeter are electrochemically accessible during potential sweeps as slow as 20 mV/s and as fast as 10 V/s. On the contrary, both redox centers along branches are accessible electrochemically within the same time frame. These results are explained in terms of slow through-space charge transfer and the globular 3-D folding of the molecules and are discussed in terms of their implications on the design of efficient redox functional dendrimers.

Leventis, Nicholas↗

Oxidation Of Manganese At Kimberley, Gale Crater: More Free Oxygen In Mars' Past?

High Mn concentrations provide unique indicators of water-rich environments and their redox state. Very high-potential oxidants are required to oxidize Mn to insoluble, high-valence oxides that can precipitate and concentrate Mn in rocks and sediments; these redox potentials are much higher than those needed to oxidize Fe or S. Consequently, Mn-rich rocks on Earth closely track the rise of atmospheric oxygen. Given the association between Mn-rich rocks and the redox state of surface environments, observations of anomalous Mn enrichments on Mars raise similar questions about redox history, solubility and aqueous transport, and availability as a metabolic substrate. Our observations suggest that at least some of the high Mn present in Gale crater occurs in the form of Mn-oxides filling veins that crosscut sand-stones, requiring post-depositional precipitation as highly oxidizing fluids moved through the fractured strata after their deposition and lithification.

Lanza, N. L.↗

Analysis of redox additive-based overcharge protection for rechargeable lithium batteries

The overcharge condition in secondary lithium batteries employing redox additives for overcharge protection, has been theoretically analyzed in terms of a finite linear diffusion model. The analysis leads to expressions relating the steady-state overcharge current density and cell voltage to the concentration, diffusion coefficient, standard reduction potential of the redox couple, and interelectrode distance. The model permits the estimation of the maximum permissible overcharge rate for any chosen set of system conditions. Digital simulation of the overcharge experiment leads to numerical representation of the potential transients, and estimate of the influence of diffusion coefficient and interelectrode distance on the transient attainment of the steady state during overcharge. The model has been experimentally verified using 1,1-prime-dimethyl ferrocene as a redox additive. The analysis of the experimental results in terms of the theory allows the calculation of the diffusion coefficient and the formal potential of the redox couple. The model and the theoretical results may be exploited in the design and optimization of overcharge protection by the redox additive approach.

Narayanan, S. R.↗

An electrochemical series of redox couples in silicate melts - A review and applications to geochemistry

An electrochemical series for redox couples in a glass-forming oxide melt is developed. This series is a quantitative numerical scale of reference reduction potentials of the redox couples in a silicate melt that is a model for basaltic magmas. The redox couples are ordered in terms of their reference reduction potentials; the order appears to be relatively independent of the exact melt composition and temperature. Thus, upon calibration to a desired composition, oxygen fugacity, and temperature, this electrochemical series can provide estimates of redox state proportions in basaltic magmas on different planetary bodies. The geochemical electrochemical series can also be used to understand the interrelationship of the redox state of the magma and the presence of volatile species such as oxygen, water, sulfur gases, and carbon gases.

Schreiber, Henry D.↗

Non-Additive Voltametric Currents From a Mixture of Two, Three and Four Redox-Active Compounds and Electroanalytical Implications

We have published recently the effect of dissimilar diffusion coefficients on the size of the voltammetric waves from a mixture of two redox-active compounds. Similarly, at the potential range where three redox-active species, decamethylferrocene (dMeFc), ferrocene (Fc) and N-methylphenothiazine (MePTZ), are oxidized simultaneously with rates controlled by linear diffusion, electrogenerated radicals diffusing outwards from the electrode react with the original species diffusing towards the electrode from the bulk; thus, Fc(+) reacts with dMeFc producing Fc and dMeFc(+), while MePTZ(+) reacts both with dMeFc producing MePTZ and dMeFc(+), and with Fc producing MePTZ and Fc(+). These reactions replace dMeFc with Fc at the second plateau, and both dMeFc and Fc with MePTZ at the third plateau. Since the diffusion coefficients of the three species are not equal, the mass-transfer limited currents of the second and the third oxidation wave plateaus change by approx. 10%. Numerical simulations of the experimental voltamograms support this mechanism. Similar results were also obtained for a mixture of four redoxactive compounds. The implications of this non-additive nature of currents on: (a) the use of internal voltammetric standards for quantitative analysis of a mixture of redox-active compounds; and, (b) the half wave potentials (E1/2) of the 2nd, 3rd and 4th waves for qualitative analysis, will be discussed.

Dass, Amala↗

Redox Bulk Energy Storage System Study, Volume 1

Opportunities were found for electrochemical energy storage devices in the U.S. electric utility industry. Application requirements for these devices were defined, including techno-economic factors. A new device, the Redox storage battery was analyzed. The Redox battery features a decoupling of energy storage and power conversion functions. General computer methods were developed to simulate Redox system operations. These studies showed that the Redox system is potentially attractive if certain performance goals can be achieved. Pathways for reducing the cost of the Redox system were identified.

Ciprios, G.↗