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NASA NTRS2010

Response of Staphylococcus Aureus to a Spaceflight Analogue

The decreased gravity of the spaceflight environment creates quiescent, low fluid shear conditions. This environment can impart considerable effects on the physiology of microorganisms as well as their interactions with potential hosts. Using the rotating wall vessel (RWV), as a spaceflight analogue, the consequence of low fluid shear culture on microbial pathogenesis has provided a better understanding of the risks to the astronaut crew from infectious microorganisms. While the outcome of low fluid shear culture has been investigated for several bacterial pathogens, little has been done to understand how this environmental factor affects Staphylococcus aureus. S. aureus is an opportunistic human pathogen which presents a high level of infection risk to the crew, as it has been isolated from both the space shuttle and International Space Station. Given that approximately forty percent of the population are carriers of the bacteria, eradication of this organism from in flight environments is impractical. These reasons have lead to us to assess the response of S. aureus to a reduced fluid shear environment. Culture in the RWV demonstrated that S. aureus grown under the low-shear condition had lower cell concentrations after 10 hours when compared to the control culture. Furthermore, the low-shear cultured bacteria displayed a reduction in carotenoid production, pigments responsible for their yellow/gold coloration. When exposed to various environmental stressors, post low-shear culture, a decrease in the ability to survive oxidative assault was observed compared to control cultures. The low fluid shear environment also resulted in a decrease in hemolysin secretion, a staphylococcal toxin responsible for red blood cell lysis. When challenged by the immune components present in human whole blood, low-shear cultured S. aureus demonstrated significantly reduced survival rates as compared to the control culture. Assays to determine the duration of these alterations demonstrated that the low-shear response could be lost in as few as 2.5 hours. These changes in phenotypic properties prompted investigation into variations occurring at the genetic level. Microarray analysis of low-shear cultured S. aureus revealed the differential regulation of genes involved in metabolism, stress response, and phosphate transfer. Additional genetic analysis with quantitative real-time PCR revealed alterations in the expression of Hfq, the conserved RNA chaperone protein involved in global gene regulation. Hfq has been connected to the regulation of a spaceflight microgravity response in S. typhimurium. These findings in S. aureus suggest an evolutionary conserved response to spaceflight conditions among structurally-diverse microorganisms. Furthermore, the reduction in pigmentation, hemolysin secretion, and survival against oxidative stress and immunologically active whole blood demonstrate an overall decrease in the virulence factors of S. aureus in response to spaceflight-like conditions.

Castro, S. L.↗

NASA NTRS2004

Transplanting Retinal Cells using Bucky Paper for Support

A novel treatment for retinal degenerative disorders involving transplantation of cells into the eye is currently under development at NASA Ames Research Center and Stanford University School of Medicine. The technique uses bucky paper as a support material for retinal pigment epithelial (RPE) cells, iris pigment epithelial (IPE) cells, and/or stem cells. This technology is envisioned as a treatment for age-related macular degeneration, which is the leading cause of blindness in persons over age 65 in Western nations. Additionally, patients with other retinal degenerative disorders, such as retinitis pigmentosa, may be treated by this strategy. Bucky paper is a mesh of carbon nanotubes (CNTs), as shown in Figure 1, that can be made from any of the commercial sources of CNTs. Bucky paper is biocompatible and capable of supporting the growth of biological cells. Because bucky paper is highly porous, nutrients, oxygen, carbon dioxide, and waste can readily diffuse through it. The thickness, density, and porosity of bucky paper can be tailored in manufacturing. For transplantation of cells into the retina, bucky paper serves simultaneously as a substrate for cell growth and as a barrier for new blood vessel formation, which can be a problem in the exudative type of macular degeneration. Bucky paper is easily handled during surgical implantation into the eye. Through appropriate choice of manufacturing processes, bucky paper can be made relatively rigid yet able to conform to the retina when the bucky paper is implanted. Bucky paper offers a distinct advantage over other materials that have been investigated for retinal cell transplantation - lens capsule and Descemet's membrane - which are difficult to handle during surgery because they are flimsy and do not stay flat.

Loftus, David J.↗

NASA NTRS2012

Development and Testing of Molecular Adsorber Coatings

The effect of on-orbit molecular contamination has the potential to degrade the performance of spaceflight hardware and diminish the lifetime of the spacecraft. For example, sensitive surfaces, such as optical surfaces, electronics, detectors, and thermal control surfaces, are vulnerable to the damaging effects of contamination from outgassed materials. The current solution to protect these surfaces is through the use of zeolite coated ceramic adsorber pucks. However, these pucks and its additional complex mounting hardware requirements result in several disadvantages, such as size, weight, and cost related concerns, that impact the spacecraft design and the integration and test schedule. As a result, a new innovative molecular adsorber coating was developed as a sprayable alternative to mitigate the risk of on-orbit molecular contamination. In this study, the formulation for molecular adsorber coatings was optimized using various binders, pigment treatment methods, binder to pigment ratios, thicknesses, and spray application techniques. The formulations that passed coating adhesion and vacuum thermal cycling tests were further tested for its adsorptive capacity. Accelerated molecular capacitance tests were performed in an innovatively designed multi-unit system containing idealized contaminant sources. This novel system significantly increased the productivity of the testing phase for the various formulations that were developed. Work performed during the development and testing phases has demonstrated successful application of molecular adsorber coatings onto metallic substrates, as well as, very promising results for the adhesion performance and the molecular capacitance of the coating. Continued testing will assist in the qualification of molecular adsorber coatings for use on future contamination sensitive spaceflight missions.

Abraham, Nithin↗

NASA NTRS2012

G-LiHT: Goddard's LiDAR, Hyperspectral and Thermal Airborne Imager

Scientists at NASA's Goddard Space Flight Center have developed an ultra-portable, low-cost, multi-sensor remote sensing system for studying the form and function of terrestrial ecosystems. G-LiHT integrates two LIDARs, a 905 nanometer single beam profiler and 1550 nm scanner, with a narrowband (1.5 nanometers) VNIR imaging spectrometer and a broadband (8-14 micrometers) thermal imager. The small footprint (approximately 12 centimeters) LIDAR data and approximately 1 meter ground resolution imagery are advantageous for high resolution applications such as the delineation of canopy crowns, characterization of canopy gaps, and the identification of sparse, low-stature vegetation, which is difficult to detect from space-based instruments and large-footprint LiDAR. The hyperspectral and thermal imagery can be used to characterize species composition, variations in biophysical variables (e.g., photosynthetic pigments), surface temperature, and responses to environmental stressors (e.g., heat, moisture loss). Additionally, the combination of LIDAR optical, and thermal data from G-LiHT is being used to assess forest health by sensing differences in foliage density, photosynthetic pigments, and transpiration. Low operating costs (approximately $1 ha) have allowed us to evaluate seasonal differences in LiDAR, passive optical and thermal data, which provides insight into year-round observations from space. Canopy characteristics and tree allometry (e.g., crown height:width, canopy:ground reflectance) derived from G-LiHT data are being used to generate realistic scenes for radiative transfer models, which in turn are being used to improve instrument design and ensure continuity between LiDAR instruments. G-LiHT has been installed and tested in aircraft with fuselage viewports and in a custom wing-mounted pod that allows G-LiHT to be flown on any Cessna 206, a common aircraft in use throughout the world. G-LiHT is currently being used for forest biomass and growth estimation in the CONUS and Mexico in support of NASA's Carbon Monitoring System (CMS) and AMIGA-Carb (AMerican Icesat Glas Assessment of Carbon). For NASA's CMS, wall-to-wall G-LiHT data have been acquired over intensive study sites with historic LiDAR datasets, dense inventory data, stem maps and flux tower observations. For AMIGA-Carb, G-LiHT transects have been acquired over ICESat tracks and USDA-FS inventory plots throughout the CONUS, and similar data will be acquired in Mexico during 2013. This talk will highlight recent science results from continental-scale transects landscape-scale deployments of G-LiHT, as well as seasonal forest dynamics from repeat pass G-LiHT acquisitions.

Cook, Bruce↗

NASA NTRS2012

John F. Kennedy Space Center's Chemochromic Hypergol Sensors

The National Aeronautics and Space Administration (NASA) seeks partne rs interested in the commercial application of the Chemochromic Hyper gol Sensors technology. NASA's Kennedy Space Center (KSC) is soliciti ng licensees for this innovative technology. The Chemochromic Hypergo l Sensors technology consists of chemochromic pigments incorporated i nto various matrices (e.g., tapes, sheets, injection molded parts, fi bers). When placed near strategic locations such as piping and contai ner valves, seams, and joints, these sensors provide an instantaneous , distinct color change from yellow to black indicating the presence of hypergols at the leak location. The chemochromic pigments can be incorporated into fibers used to make fabrics for personal protective equipment as well as into badge holders for use as a point leak detector. These affordable, easily replaceable sensors provide the capabil ity to visually monitor leak-prone locations and personnel working i n those areas on a continuous basis for the presence of dangerous hyp ergols.

Nichols, James D.↗

NASA NTRS2013

Color-Changing Sensors for Detecting the Presence of Hypergolic Fuels

Hypergolic fuel sensors were designed to incorporate novel chemochromic pigments into substrates for use in various methods of leak detection. There are several embodiments to this invention that would provide specific visual indication of hypergols used during and after transfer. The ability to incorporate these pigments into various polymer matrices provides a unique opportunity to manufacture nearly any type of sensor shape that is required. The vibrant color change from yellow to black instantaneously shows the worker the presence of hypergols in the area.

Roberson, Luke↗

NASA NTRS2014

Black Molecular Adsorber Coatings for Spaceflight Applications

The molecular adsorber coating is a new technology that was developed to mitigate the risk of on-orbit molecular contamination on spaceflight missions. The application of this coating would be ideal near highly sensitive, interior surfaces and instruments that are negatively impacted by outgassed molecules from materials, such as plastics, adhesives, lubricants, epoxies, and other similar compounds. This current, sprayable paint technology is comprised of inorganic white materials made from highly porous zeolite. In addition to good adhesion performance, thermal stability, and adsorptive capability, the molecular adsorber coating offers favorable thermal control characteristics. However, low reflectivity properties, which are typically offered by black thermal control coatings, are desired for some spaceflight applications. For example, black coatings are used on interior surfaces, in particular, on instrument baffles for optical stray light control. Similarly, they are also used within light paths between optical systems, such as telescopes, to absorb light. Recent efforts have been made to transform the white molecular adsorber coating into a black coating with similar adsorptive properties. This result is achieved by optimizing the current formulation with black pigments, while still maintaining its adsorption capability for outgassing control. Different binder to pigment ratios, coating thicknesses, and spray application techniques were explored to develop a black version of the molecular adsorber coating. During the development process, coating performance and adsorption characteristics were studied. The preliminary work performed on black molecular adsorber coatings thus far is very promising. Continued development and testing is necessary for its use on future contamination sensitive spaceflight missions.

adsorption↗

NASA NTRS2014

Black Molecular Adsorber Coatings for Spaceflight Applications

adsorption↗

NASA NTRS2015

Chemochromic Detector for Sensing Gas Leakage and Process for Producing the Same

A chemochromic sensor for detecting a combustible gas, such as hydrogen, includes a chemochromic pigment and a textile polymer. The textile material includes a chemochromic pigment operably responsive to a combustible gas. The combustible gas sensing textile material can be made by melt spinning, solution spinning, or other similar techniques. In a preferred embodiment carbon nanotubes are used with the textile material which will increase the material strength and alter the thermal and/or electrical properties. These textiles woven into fabrics can provide garments not only with hydrogen sensing capabilities but the carbon nanotubes will allow for a range of sensing capabilities to be embedded (i.e. gas, health, and electronic monitors) within the garments.

Roberson, Luke B.↗

NASA NTRS2015

Spectral and Temporal Laser Fluorescence Analysis Such as for Natural Aquatic Environments

An Advanced Laser Fluorometer (ALF) can combine spectrally and temporally resolved measurements of laser-stimulated emission (LSE) for characterization of dissolved and particulate matter, including fluorescence constituents, in liquids. Spectral deconvolution (SDC) analysis of LSE spectral measurements can accurately retrieve information about individual fluorescent bands, such as can be attributed to chlorophyll-a (Chl-a), phycobiliprotein (PBP) pigments, or chromophoric dissolved organic matter (CDOM), among others. Improved physiological assessments of photosynthesizing organisms can use SDC analysis and temporal LSE measurements to assess variable fluorescence corrected for SDC-retrieved background fluorescence. Fluorescence assessments of Chl-a concentration based on LSE spectral measurements can be improved using photo-physiological information from temporal measurements. Quantitative assessments of PBP pigments, CDOM, and other fluorescent constituents, as well as basic structural characterizations of photosynthesizing populations, can be performed using SDC analysis of LSE spectral measurements.

Chekalyuk, Alexander↗

NASA NTRS2014

Estimation of Crop Gross Primary Production (GPP): Fapar(sub Chl) Versus MOD15A2 FPAR

Photosynthesis (PSN) is a pigment level process in which antenna pigments (predominately chlorophylls) in chloroplasts absorb photosynthetically active radiation (PAR) for the photochemical process. PAR absorbed by foliar non-photosynthetic components is not used for PSN. The fraction of PAR absorbed (fAPAR) by a canopy/vegetation (i.e., fAPAR(sub canopy)) derived from the Moderate Resolution Imaging Spectroradiometer (MODIS) images, referred to as MOD15A2 FPAR, has been used to compute absorbed PAR (APAR) for PSN (APARPSN) which is utilized to produce the standard MODIS gross primary production (GPP) product, referred to as MOD17A2 GPP. In this study, the fraction of PAR absorbed by chlorophyll throughout the canopy (fAPAR(sub chl)) was retrieved from MODIS images for three AmeriFlux crop fields in Nebraska. There are few studies in the literature that compare the performance of MOD15A2 FPAR versus fAPAR(sub chl) in GPP estimation. In our study MOD15A2 FPAR and the retrieved fAPAR(sub chl) were compared with field fAPAR(sub canopy) and the fraction of PAR absorbed by green leaves of the vegetation (fAPAR(sub green)). MOD15A2 FPAR overestimated field fAPAR(sub canopy) in spring and in fall, and underestimated field fAPAR(sub canopy) in midsummer whereas fAPAR(sub chl) correctly captured the seasonal phenology. The retrieved fAPAR(sub chl) agreed well with field fAPAR(sub green) at early crop growth stage in June, and was less than field fAPAR(sub green) in late July, August and September. GPP estimates with fAPAR(sub chl) and with MOD15A2 FPAR were compared to tower flux GPP. GPP simulated with fAPAR(sub chl) was corroborated with tower flux GPP. Improvements in crop GPP estimation were achieved by replacing MOD15A2 FPAR with fAPAR(sub chl) which also reduced uncertainties of crop GPP estimates by 1.12-2.37 g C m(sup −2) d(sup −1).

GPM↗

NASA NTRS2015

Color Changing Hydrogen Sensors

During the Space Shuttle Program, one of the most hazardous operation that occurred was the loading of liquid hydrogen (LH2) during fueling operations of the spacecraft. Due to hydrogen's low explosive limit, any amount leaked could lead to catastrophic event. Hydrogen's chemical properties make it ideal as a rocket fuel; however, the fuel is deemed unsafe for most commercial use because of the inability to easily detect the gas leaking. The increased use of hydrogen over traditional fossil fuels would reduce greenhouse gases and America's dependency on foreign oil. Therefore a technology that would improve safety at NASA and in the commercial sector while creating a new economic sector would have a huge impact to NASA's mission. The Chemochromic Detector for sensing hydrogen gas leakage is a color-changing detector that is useful in any application where it is important to know not only the presence but also the location of the hydrogen gas leak. This technology utilizes a chemochromicpigment and polymer matrix that can be molded or spun into rigid or pliable shapes useable in variable temperature environments including atmospheres of inert gas, hydrogen gas, or mixtures of gases. A change in color of the detector material indicates where gaseous hydrogen leaks are occurring. The irreversible sensor has a dramatic color change from beige to dark grey and remains dark grey after exposure. A reversible pigment changes from white to blue in the presence of hydrogen and reverts back to white in the presence of oxygen. Both versions of the sensor's pigments were comprised of a mixture of a metal oxide substrate and a hydro-chromic compound (i.e., the compound that changed color in the presence of hydrogen) and immediately notified the operator of the presence of low levels of hydrogen. The detector can be used in a variety of formats including paint, tape, caulking, injection molded parts, textiles and fabrics, composites, and films. This technology brings numerous benefits over the traditional hydrogen sensors: The technology has excellent temperature stability (4K to 373 K), it can be used in cryogenic fluid applications, it is easy to apply and remove; it requires no power to operate; it has a quick response time; the leak points can be detected visually or electronically; it is nonhazardous, thus environmentally friendly; it can be reversible or irreversible; it does not require on-site monitoring; has a long shelf life; the detector is very durable; and the technology is inexpensive to manufacture.

Chemochromic sensor↗

NASA NTRS2019

Applicability of Spectral Decomposition by Varimax-Rotated, Principal Component Analysis to the Surface Biology and Geology (SBG) VNIR Mission Concept

Cyanobacterial and Harmful Algal Blooms (CyanoHABs) are a growing concern in coastal and inland waters. But, spectral interference from multiple constituents in optically complex waters can hamper application of remote sensing using traditional image processing methods. The Kent State University (KSU) spectral decomposition method can be applied to multispectral and hyperspectral remote sensing images (e.g. HICO and the NASA Glenn HSI2) to partition and identify signals related to cyanobacteria, algae, pigment degradation products and suspended sediment in each pixel. Fundamental to the use of remote sensing data is the ability to extract independent signals from correlated hyperspectral VNIR data cubes. The Kent State University varimax-rotated, principal component analysis method (VPCA) is important to integrate into the SBG VNIR mission concept because it provides greater specificity, a software-based SNR boost relative to hardware performance, and can assist with Cal/Val, Modeling and Applications. We present examples of the hyperspectral application of the KSU VPCA method with relevance to SBG. The information extracted by VPCA can be validated spectrally or spatially with laboratory and/or in situ sensors, which capture spatial or time series of information at discrete points within remote sensing images. Comparisons show hyperspectral sensors extract more components than multispectral ones, but more independent information can be extracted from multispectral sensors by VPCA than traditional band ratio approaches. The spectral decomposition method is capable of enhancing the signal to noise ratio (SNR) of the NASA Glenn, second-generation hyperspectral imager by a factor of 7x to 20x, with a spectral reproducibility of ±3%. The spectral decomposition method, when compared against existing remote sensing monitoring methods exhibits both greater specificity and a lower detection limit. The method has been validated with multispectral images in Lake Erie to quantify the Microcystis CyanoHAB and from the Indian River Lagoon, Florida to quantify the Brown Tide resulting from A. lagunesnsis. Field operations in the Western Basin of Lake Erie were conducted using a bbe Fluoroprobe to collect vertical profiles and horizontal tows along a transect from the Toledo to the Detroit Lighthouse during coincident satellite overpasses. Extraction of pixel values from the MODIS Aqua sensor yields agreement between in situ field and lab-based measures of cyanobacterial, cryptophyte, diatoms and green algae, suspended sediment and pigment degradation products with R2>0.8.

hyperspectral↗

NASA NTRS2021

Cryovolcanic flooding in Viking Terra on Pluto

A prominent fossa trough (Uncama Fossa) and adjacent 28-km diameter impact crater (Hardie) in Pluto's Viking Terra, as seen in the high-resolution images from the New Horizons spacecraft, show morphological evidence of in-filling with a material of uniform texture and red-brown color. A linear fissure parallel to the trough may be the source of a fountaining event yielding a cryoclastic deposit having the same composition and color properties as is found in the trough and crater. Spectral maps of this region with the New Horizons LEISA instrument reveal the spectral signature of H2O ice in these structures and in distributed patches in the adjacent terrain in Viking Terra. A detailed statistical analysis of the spectral maps shows that the colored H2O ice filling material also carries the 2.2-μm signature of an ammoniated component that may be an ammonia hydrate (NH3·nH2O) or an ammoniated salt. This paper advances the view that the crater and fossa trough have been flooded by a cryolava debouched from Pluto's interior along fault lines in the trough and in the floor of the impact crater. The now frozen cryolava consisted of liquid H2O infused with the red-brown pigment presumed to be a tholin, and one or more ammoniated compounds. Although the abundances of the pigment and ammoniated compounds entrained in, or possibly covering, the H2O ice are unknown, the strong spectral bands of the H2O ice are clearly visible. In consideration of the factors in Pluto's space environment that are known to destroy ammonia and ammonia-water mixtures, the age of the exposure is of order ≤109 years. Ammoniated salts may be more robust, and laboratory investigations of these compounds are needed.

Pluto↗

NASA NTRS2021

Impact of Spectral Resolution on Quantifying Cyanobacteria in Lakes and Reservoirs: A Machine-Learning Assessment

Cyanobacterial harmful algal blooms are an increasing threat to coastal and inland waters. These blooms can be detected using optical radiometers due to the presence of phycocyanin (PC) pigments. The spectral resolution of best-available multispectral sensors limits their ability to diagnostically detect PC in the presence of other photosynthetic pigments. To assess the role of spectral resolution in the determination of PC, a large ( N=905 ) database of colocated in situ radiometric spectra and PC are employed. We first examine the performance of selected widely used machine-learning (ML) models against that of benchmark algorithms for hyperspectral remote sensing reflectance ( R_(rs) ) spectra resampled to the spectral configuration of the Hyperspectral Imager for the Coastal Ocean (HICO) with a full-width at half-maximum (FWHM) of < 6 nm. Results show that the multilayer perceptron (MLP) neural network applied to HICO spectral configurations (median errors < 65%) outperforms other ML models. This model is subsequently applied to R_(rs) spectra resampled to the band configuration of existing satellite instruments and of the one proposed for the next Landsat sensor. These results confirm that employing MLP models to estimate PC from hyperspectral data delivers tangible improvements compared with retrievals from multispectral data and benchmark algorithms (with median errors between ∼73 % and 126%) and shows promise for developing a globally applicable cyanobacteria measurement approach.

hyperspectral↗

DOE OSTI2025

Functional Connectivity of Red Chlorophylls in Cyanobacterial Photosystem I Revealed by Fluence-Dependent Transient Absorption

External stressors modulate the oligomerization state of photosystem I (PSI) in cyanobacteria. The number of red chlorophylls (Chls), pigments lower in energy than the P700 reaction center, depends on the oligomerization state of PSI. Here, we use ultrafast transient absorption spectroscopy to interrogate the effective connectivity of the red Chls in excitonic energy pathways in trimeric PSI in native thylakoid membranes of the model cyanobacterium Synechocystis sp. PCC 6803, including emergent dynamics, as red Chls increase in number and proximity. Fluence-dependent dynamics indicate singlet–singlet annihilation within energetically connected red Chl sites in the PSI antenna but not within bulk Chl sites on the picosecond time scale. These data support picosecond energy transfer between energetically connected red Chl sites as the physical basis of singlet–singlet annihilation. The time scale of this energy transfer is faster than predicted by Förster resonance energy transfer calculations, raising questions about the physical mechanism of the process. Our results indicate distinct strategies to steer excitations through the PSI antenna; the red Chls present a shallow reservoir that direct excitations away from P 700 , extending the time to trapping by the reaction center.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2024

Controlling Vibronic Coupling in Chlorophyll Proteins: The Effects of Excitonic Delocalization and Vibrational Localization

Vibrational-electronic (vibronic) coupling plays a critical role in excitation energy transfer in molecular aggregates and pigment-protein complexes (PPCs). But the interplay between excitonic delocalization and vibronic interactions is complex, often leaving even qualitative questions as to what conceptual framework (e.g., Redfield versus Forster theory) should be used to interpret experimental results. To shed light on this issue, we report here on the interplay between excitonic delocalization and vibronic coupling in site-directed mutants of the water soluble chlorophyll protein (WSCP), as reflected in 77 K fluorescence spectra. Experimentally, we find that in PPCs where excitonic delocalization is disrupted (either by mutagenesis or heterodimer formation), the relative intensity of the vibrational sideband (VSB) in fluorescence spectra is suppressed by up to 37% compared to the native protein. Furthermore, numerical simulations reveal that this effect results from the localization of high-frequency vibrations in the coupled system; while excitonic delocalization suppresses the purely electronic transition due to H-aggregate-like dipole-dipole interference, high-frequency vibrations are unaffected, leading to a relative enhancement of the VSB. By comparing VSB intensities of PPCs both in the presence and absence of excitonic delocalization, we extract a set of “local” Huang-Rhys (HR) factors for Chl a in WSCP. More generally, our results suggest a significant role for geometric effects in controlling energy-transfer rates (which depend sensitively on absorption/fluorescence line shapes) in molecular aggregates and PPCs.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2026

Phycocyanobilin biosynthesis in Galdieria sulphuraria requires isomerization of phycoerythrobilin synthesized by bilin reductases

Phycobiliproteins are essential components of the light-harvesting antennae in cyanobacteria and red algae, requiring covalently bound open-chain tetrapyrrole chromophores (bilins) for proper function. In the red alga Galdieria sulphuraria , the primary chromophore is phycocyanobilin (PCB), despite the apparent presence of only biosynthetic genes for phycoerythrobilin (PEB) biosynthesis (PEBA and PEBB). This observation suggests the presence of an alternative, atypical biosynthetic pathway for PCB. In this study, we confirmed the presence of PEB:PCB isomerase activity in an enriched protein fraction from G. sulphuraria . To further investigate this unusual pathway, we combined in silico analyses with biochemical assays. Phylogenetic analyses confirmed the placement of the G. sulphuraria ferredoxin-dependent bilin reductases within the PEBA and PEBB lineages, typically associated with PEB synthesis, whereas the related red alga Cyanidioschyzon merolae was found to contain only PCYA. This gene distribution presents a functional paradox. G. sulphuraria harbors PEB biosynthesis genes but no detectable PEB chromophores and lacks known PCB-synthesizing enzymes despite containing PCB. Functional characterization of recombinant GsPEBA (G. sulphuraria PEBA) and GsPEBB ( G. sulphuraria PEBB) confirmed their roles in PEB synthesis, demonstrating that these enzymes have not evolved to synthesize PCB or act as isomerases despite their phylogenetic placement. In contrast, Cm PCYA ( C. merolae PCYA) catalyzed direct PCB formation from biliverdin. Together, these findings reveal an atypical isomerase-based pathway for PCB biosynthesis in G. sulphuraria , expanding our understanding of bilin metabolism and providing new insight into the evolutionary flexibility of photosynthetic pigment biosynthesis in Rhodophyta.

14 SOLAR ENERGY↗