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All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Proteomes reveal metabolic capabilities of Yarrowia lipolytica for biological upcycling of polyethylene into high-value chemicals

ABSTRACT Polyolefins derived from plastic wastes are recalcitrant for biological upcycling. However, chemical depolymerization of polyolefins can generate depolymerized plastic (DP) oil, comprising of a complex mixture of saturated, unsaturated, even, and odd hydrocarbons suitable for biological conversion. While DP oil contains a rich carbon and energy source, it is inhibitory to cells. Understanding and harnessing robust metabolic capabilities of microorganisms to upcycle the hydrocarbons in DP oil, both naturally and unnaturally occurring, into high-value chemicals are limited. Here, we discovered that an oleaginous yeast, Yarrowia lipolytica, undergoing short-term adaptation to DP oil robustly utilized a wide range of hydrocarbons for cell growth and production of citric acid and neutral lipids. When growing on hydrocarbons, Y. lipolytica partitioned into planktonic and oil-bound cells with each exhibiting distinct proteomes and amino acid distributions invested in establishing these proteomes. Significant proteome reallocation toward energy and lipid metabolism, belonging to 2 of the 23 Eukaryotic Orthologous Groups classes C and I, enabled robust growth of Y. lipolytica on hydrocarbons, with n-hexadecane as the preferential substrate. This investment was even higher for growth on DP oil where classes C and I were ranked one and two, respectively, and many associated proteins and pathways were expressed and upregulated including the hydrocarbon degradation pathway, Krebs cycle, glyoxylate shunt and, unexpectedly, propionate metabolism. However, a reduction in proteome allocation for protein biosynthesis, at the expense of the observed increase toward energy and lipid metabolisms, might have caused the inhibitory effect of DP oil on cell growth. IMPORTANCE Sustainable processes for biological upcycling of plastic wastes in a circular bioeconomy are needed to promote decarbonization and reduce environmental pollution due to increased plastic consumption, incineration, and landfill storage. Strain characterization and proteomic analysis revealed the robust metabolic capabilities of Yarrowia lipolytica to upcycle polyethylene into high-value chemicals. Significant proteome reallocation toward energy and lipid metabolisms was required for robust growth on hydrocarbons with n-hexadecane as the preferential substrate. However, an apparent over-investment in these same categories to utilize complex depolymerized plastic (DP) oil came at the expense of protein biosynthesis, limiting cell growth. Taken together, this study elucidates how Y. lipolytica activates its metabolism to utilize DP oil and establishes Y. lipolytica as a promising host for the upcycling of plastic wastes.

1-alkenes↗
DOE OSTI2023

Comparative Genomic Analyses of Cellulolytic Machinery Reveal Two Nutritional Strategies of Marine Labyrinthulomycetes Protists

Labyrinthulomycetes are a group of ubiquitous and diverse unicellular Stramenopiles and have long been known for their vital role in ocean carbon cycling. However, their ecological function from the perspective of organic matter degradation remains poorly understood. This study reports high-quality genomes of two newly isolated Labyrinthulomycetes strains, namely, Botryochytrium sp. strain S-28 and Oblongichytrium sp. strain S-429, and provides molecular analysis of their ecological functions using comparative genomics and a biochemical assay. Our results suggest that Labyrinthulomycetes may occupy multiple ecological niches in marine ecosystems because of the significant differences in gene function among different genera. Certain strains could degrade wheat bran independently by secreting cellulase. The key glycoside hydrolase families (GH1, GH5, and GH9) related to cellulase and the functional domains of carbohydrate-active enzymes (CAZymes) were more enriched in their genomes. This group can actively participate in marine biochemical cycles as decomposers. In contrast, other strains that could not produce cellulase may thrive as “leftover scavengers” and act as a source of nutrients to the higher-trophic-level plankton. In addition, our findings emphasize the dual roles of endoglucanase, acting as both exo- and endoglucanases, in the process of cellulose degradation. Using genomic, biochemical, and phylogenetic analyses, our study provides a broader insight into the nutritional patterns and ecological functions of Labyrinthulomycetes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

A green sulfur bacterium from epsomitic Hot Lake, Washington, USA

Hot Lake is a small heliothermal and hypersaline lake in far north–central Washington State (USA) and is limnologically unusual because MgSO 4 rather than NaCl is the dominant salt. In late summer, the Hot Lake metalimnion becomes distinctly green from blooms of planktonic phototrophs. In a study undertaken over 60 years ago, these blooms were predicted to include green sulfur bacteria, but no cultures were obtained. We sampled Hot Lake and established enrichment cultures for phototrophic sulfur bacteria in MgSO 4 -rich sulfidic media. Most enrichments turned green or red within 2 weeks, and from green-colored enrichments, pure cultures of a lobed green sulfur bacterium (phylum Chlorobi) were isolated. Phylogenetic analyses showed the organism to be a species of the prosthecate green sulfur bacterium Prosthecochloris. Cultures of this Hot Lake phototroph were halophilic and tolerated high levels of sulfide and MgSO 4 . In addition, unlike all recognized species of Prosthecochloris, the Hot Lake isolates grew at temperatures up to 45°C, indicating an adaptation to the warm summer temperatures of the lake. Photoautotrophy by Hot Lake green sulfur bacteria may contribute dissolved organic matter to anoxic zones of the lake, and their diazotrophic capacity may provide a key source of bioavailable nitrogen, as well.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

DVRFS Microbiome 2021 Manuscript Code

These scripts document the code used to analyze the Death Valley Regional Flow System (DVRFS) microbial community for a manuscript titled "Subsurface Planktonic Microbial Communities Reflect Regional-Scale Groundwater Hydraulic Connectivity".

Merino, NancyS.↗
DOE OSTI2020

A pipeline for targeted metagenomics of environmental bacteria

Background:Metagenomics and single cell genomics provide a window into the genetic repertoire of yet uncultivated microorganisms, but both methods are usually taxonomically untargeted. The combination of fluorescence in situ hybridization (FISH) and fluorescence activated cell sorting (FACS) has the potential to enrich taxonomically well-defined clades for genomic analyses. Methods:Cells hybridized with a taxon-specific FISH probe are enriched based on their fluorescence signal via flow cytometric cell sorting. A recently developed FISH procedure, the hybridization chain reaction (HCR)-FISH, provides the high signal intensities required for flow cytometric sorting while maintaining the integrity of the cellular DNA for subsequent genome sequencing. Sorted cells are subjected to shotgun sequencing, resulting in targeted metagenomes of low diversity. Results: Pure cultures of different taxonomic groups were used to (1) adapt and optimize the HCR-FISH protocol and (2) assess the effects of various cell fixation methods on both the signal intensity for cell sorting and the quality of subsequent genome amplification and sequencing. Best results were obtained for ethanol-fixed cells in terms of both HCR-FISH signal intensity and genome assembly quality. Our newly developed pipeline was successfully applied to a marine plankton sample from the North Sea yielding good quality metagenome assembled genomes from a yet uncultivated flavobacterial clade. Conclusions: With the developed pipeline, targeted metagenomes at various taxonomic levels can be efficiently retrieved from environmental samples. The resulting metagenome assembled genomes allow for the description of yet uncharacterized microbial clades.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Microbial community dynamics and coexistence in a sulfide-driven phototrophic bloom

Background: Lagoons are common along coastlines worldwide and are important for biogeochemical element cycling, coastal biodiversity, coastal erosion protection and blue carbon sequestration. These ecosystems are frequently disturbed by weather, tides, and human activities. Here, we investigated a shallow lagoon in New England. The brackish ecosystem releases hydrogen sulfide particularly upon physical disturbance, causing blooms of anoxygenic sulfur-oxidizing phototrophs. To study the habitat, microbial community structure, assembly and function we carried out in situ experiments investigating the bloom dynamics over time. Results: Phototrophic microbial mats and permanently or seasonally stratified water columns commonly contain multiple phototrophic lineages that coexist based on their light, oxygen and nutrient preferences. We describe similar coexistence patterns and ecological niches in estuarine planktonic blooms of phototrophs. The water column showed steep gradients of oxygen, pH, sulfate, sulfide, and salinity. The upper part of the bloom was dominated by aerobic phototrophic Cyanobacteria, the middle and lower parts by anoxygenic purple sulfur bacteria (Chromatiales) and green sulfur bacteria (Chlorobiales), respectively. We show stable coexistence of phototrophic lineages from five bacterial phyla and present metagenome-assembled genomes (MAGs) of two uncultured Chlorobaculum and Prosthecochloris species. In addition to genes involved in sulfur oxidation and photopigment biosynthesis the MAGs contained complete operons encoding for terminal oxidases. The metagenomes also contained numerous contigs affiliating with Microviridae viruses, potentially affecting Chlorobi. Our data suggest a short sulfur cycle within the bloom in which elemental sulfur produced by sulfide-oxidizing phototrophs is most likely reduced back to sulfide by Desulfuromonas sp. Conclusions: The release of sulfide creates a habitat selecting for anoxygenic sulfur-oxidizing phototrophs, which in turn create a niche for sulfur reducers. Strong syntrophism between these guilds apparently drives a short sulfur cycle that may explain the rapid development of the bloom. The fast growth and high biomass yield of Chlorobi-affiliated organisms implies that the studied lineages of green sulfur bacteria can thrive in hypoxic habitats. This oxygen tolerance is corroborated by oxidases found in MAGs of uncultured Chlorobi. The findings improve our understanding of the ecology and ecophysiology of anoxygenic phototrophs and their impact on the coupled biogeochemical cycles of sulfur and carbon.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Marine aerosol generation experiments in the High Arctic during summertime

The rapidly warming Arctic has transitioned to thinner sea ice which fractures, producing leads. Few studies have investigated Arctic sea spray aerosol (SSA) produced from open ocean, leads, and melt ponds, which vary in salinity and organic and microbial community composition. A marine aerosol reference tank was deployed aboard an icebreaker to the Arctic Ocean during August–September 2018 to study SSA generated from locally collected surface waters. Aerosol generation experiments were carried out using water collected from the marginal ice zone, a human-made hole in sea ice near the North Pole, and both lead and melt pond water during an ice floe drift period. Salinity, chlorophyll a, organic carbon, nitrogen, and microbial community composition were measured. Eukaryotic plankton and bacterial abundance were elevated in experimental water from the marginal ice zone, but the relative contributions from major eukaryotic taxonomic groups varied little across the experiments. The chemical composition of individual SSA particles was analyzed using Raman microspectroscopy and computer-controlled scanning electron microscopy with energy-dispersive X-ray spectroscopy. Individual sea salt aerosol, primary organic aerosol, and mineral dust particles were observed. Sea salt aerosol constituted 44–95% of individual submicrometer and 68–100% of supermicrometer particles, by number, generated during each experiment. Carbon was detected in 85%, by number, of the individual sea salt particles, with visible organic coatings. Carbohydrates were detected in 72% of particles, by number, with smaller contributions from long-chain fatty acids (13%) and siliceous material (15%). SSA generated from melt pond water contained only long-chain fatty acids and siliceous material. Quantification of the ice-nucleating activity showed that locally produced SSA may define the High Arctic background ice-nucleating particle population, but cannot account for the peak atmospheric concentrations observed. As the Arctic warms, the increasing SSA emissions have a complex dependence on changing biological and physical processes.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Quantitative Biostratigraphic Analysis and Age Estimates of Middle Cretaceous Sequences in The Baltimore Canyon Trough, Offshore Mid-Atlantic U.S. Margin

ABSTRACT We applied quantitative methods to previously published biostratigraphic data from the Baltimore Canyon Trough (offshore of the Mid-Atlantic U.S.A.) to provide an improved chronostratigraphic framework for Cretaceous sequences. Here, we successfully used graphic correlation of 228 planktonic foraminifera, nannofossil, and palynological events spanning 22 wells to define assemblage and interval zones as well as major paleoenvironmental changes in the Dawson Canyon, Logan Canyon (three sequences), and Missisauga Formations (two sequences, undifferentiated here). Ranking and scaling techniques were not successful because of the of the limited number of usable biostratigraphic markers. The ages of the sequences previously identified using well logs and seismic profiles were temporally constrained based on chronostratigraphically significant biostratigraphic markers that we identified: the late Cenomanian to Turonian DCx sequence (Rotalipora cushmani and Thalmanninella greenhornensis); the early Cenomanian LC1 sequence; the middle and late Albian LC2 sequence (Braarudosphaera africana, Planomalina buxtorfi, and Spinidinium vestitum); the late Aptian LC3 sequence (Cyclonephelium tabulatum); and the early Aptian to Barremian Missisauga sequences (Aptea anaphrissa, Pseudoceratium pelliferum, and Muderongia simplex). These five biostratigraphic associations are correlated with six prominent seismic reflectors and sequence boundaries that can be traced across the basin. Duration of hiatuses associated with these sequence boundaries are uncertain, though our Monte Carlo analysis allows extraction of age estimates from broad and sometimes contradictory ranges and suggests correlation of hiatuses with global sea-level falls. Together, these seismic and biostratigraphic interpretations can be applied (1) to evaluate reservoir continuity and the viability of offshore carbon storage reservoirs in the Baltimore Canyon Trough, (2) to better define the tectonostratigraphic evolution of the basin, and (3) to contribute to the understanding of regional and global variations in Cretaceous sea level.

Paleontology↗
DOE OSTI2021

Analysis of microplastics in bivalves along Fourmile Branch

Microplastics are commonly found near wastewater treatment facilities with the source originating typically from fibers associated with laundry detergents. Fourmile branch would have limited laundry associated effluent and does not receive any input from water originating from upstream industry sources, therefore microplastics would have originated from Site operations or through atmospheric deposition which are both unexplored pathways. In order to assess environmental inventory effects on the biota, water samples were collected from sampling locations along Fourmile Branch on the Savannah River Site using plankton nets and grab samples at Fourmile Branch locations (i.e., FM-2B, FM-A7, and FM6). Fourmile Branch has a long history of receiving industrial effluents from site operations as well effluent from the site’s wastewater treatment plant. Water samples and the debris collected in the nets were rinsed with deionized water and sieved to remove the larger fractions of plastics (4000-2000 µm) and retain fractions <500 µm. The water samples were analyzed for the type and size of plastics by μ-Raman, mass spectrometry, and Fourier Transform Infrared Spectroscopy (FTIR) methods. Previously collected bivalves were prepared into thin sections and analyzed using microscopy.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2025

Biophysical model of eelgrass and water quality in Coos Bay, OR shows greater mitigation potential for ocean acidification than hypoxia

Seagrass beds provide important ecosystem services and are valued, in part, for their potential to mediate stressors such as ocean acidification and hypoxia (OAH) for sensitive species. However, the susceptibility of seagrasses to anthropogenic impacts and recent declines motivate the need to better understand the drivers of seagrass and the water quality consequences that occur with variation in seagrass abundance. To meet this need, we leveraged existing monitoring data (water quality and seagrass), hydrodynamic circulation model, and biogeochemical model framework with seagrass submodel, to produce a biophysical model of Coos Bay estuary, Oregon, U.S. The model includes biogeochemical processes involving water quality, plankton, seagrass, and sediment-water interactions. Ecosystem models like this are useful for evaluating complex estuarine systems because they allow us to extend our understanding of system dynamics beyond existing observations and perform experiments to identify the processes driving observed patterns. We used the biophysical model of Coos Bay to evaluate the dynamics of water quality and native eelgrass (Zostera marina) under three eelgrass abundance scenarios (zero eelgrass, current extent, and maximum observed extent) to elucidate the relationship between eelgrass and OAH. Including eelgrass in the Coos Bay model produced results that more closely resembled water quality observations - dissolved oxygen (DO) and pH were more dynamic in simulations with eelgrass, often having both higher highs and lower lows. While there were some areas of the estuary where DO improved with the addition of eelgrass to the model there was overall a small net increase in harmful DO conditions (based on a salmon physiological threshold). In contrast, ocean acidification conditions, pH and calcium carbonate saturation state for aragonite (Ω), were improved (based on oyster requirements) with the addition of eelgrass - although the magnitude of improvement differed seasonally and spatially. Our new model represents a useful tool - one which accounts for and controls the relevant physical and biogeochemical processes - to evaluate conditions that confer resilience or enhance vulnerability to OAH in an important Pacific Northwest coastal estuary and results can inform the OAH-related dynamics occurring in other eastern boundary current estuaries.

FVCOM-ICM↗
DOE OSTI2025

Ocean liming effect on a North Atlantic microbial community: changes in composition and rates

The ongoing rise in atmospheric CO 2 levels and the consequent global warming make it increasingly difficult to maintain the global temperature within the 1.5 - 2°C target set by the Paris Agreement. Therefore, strategies to remove carbon dioxide from the atmosphere are being developed, with ocean alkalinity enhancement (OAE) gaining most attention. Within OAE, ocean liming- the addition of quicklime (CaO) or hydrated lime (Ca(OH) 2 )- can not only remove CO 2 from the atmosphere but potentially counteract the effects of ocean acidification. Although quite attractive, these technologies have yet to be tested regarding ecological safety and efficacy. Here we report the impacts of ocean liming on the abundance, composition and extracellular enzymatic activity (EEA) rates of a North Atlantic planktonic community. The results demonstrate that OAE led to a decreased phytoplankton development, mainly diatoms. The bacterial response to OAE was community-specific, with a consistent increase in the relative abundance of the order Oceanospirillales. OAE also led to increased EEA rates, especially within the bacterial community. These findings suggest that while initial effects on phytoplankton may be limited, the specific impacts on bacterial groups suggest that OAE could influence the remineralization of organic matter. If our results apply to other communities, OAE might initially affect marine microbial dynamics, but further studies are needed to determine if these effects are long-term.

16S↗
DOE OSTI2020

PcsR2 Is a LuxR-Type Regulator That Is Upregulated on Wheat Roots and Is Unique to Pseudomonas chlororaphis

LuxR solos are common in plant-associated bacteria and increasingly recognized for playing important roles in plant-microbe interkingdom signaling. Unlike the LuxR-type transcriptional regulators of prototype LuxR/LuxI quorum sensing systems, luxR solos do not have a LuxI-type autoinducer synthase gene associated with them. LuxR solos in plant-pathogenic bacteria are important for virulence and in plant endosymbionts contribute to symbiosis. In the present study, we characterized an atypical LuxR solo, PcsR2, in the biological control species Pseudomonas chlororaphis 30–84 that is highly conserved among sequenced P. chlororaphis strains. Unlike most LuxR solos in the plant-associated bacteria characterized to date, pcsR2 is not associated with a proline iminopeptidase gene and the protein has an atypical N-terminal binding domain. We created a pcsR2 deletion mutant and used quantitative RT-PCR to show that the expression of pcsR2 and genes in the operon immediately downstream was upregulated ~10-fold when the wild type strain was grown on wheat roots compared to planktonic culture. PcsR2 was involved in upregulation. Using a GFP transcriptional reporter, we found that expression of pcsR2 responded specifically to root-derived substrates as compared to leaf-derived substrates but not to endogenous AHLs. Compared to the wild type, the mutant was impaired in the ability to utilize root carbon and nitrogen sources in wheat root macerate and to colonize wheat roots. Phenazine production and most biofilm traits previously shown to be correlated with phenazine production also were diminished in the mutant. Gene expression of several of the proteins in the phenazine regulatory network including PhzR, Pip (phenazine inducing protein) and RpeA/RpeB were reduced in the mutant, and overexpression of these genes in trans restored phenazine production in the mutant to wild-type levels, indicating PcsR2 affects the activity of the these regulatory genes upstream of RpeA/RpeB via an undetermined mechanism. Our results indicate PcsR2 upregulates the expression of the adjacent operon in response to unknown wheat root-derived signals and belongs to a novel subfamily of LuxR-type transcriptional regulators found in sequenced P. chlororaphis strains.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Antarctic lake viromes reveal potential virus associated influences on nutrient cycling in ice-covered lakes

The McMurdo Dry Valleys (MDVs) of Antarctica are a mosaic of extreme habitats which are dominated by microbial life. The MDVs include glacial melt holes, streams, lakes, and soils, which are interconnected through the transfer of energy and flux of inorganic and organic material via wind and hydrology. For the first time, we provide new data on the viral community structure and function in the MDVs through metagenomics of the planktonic and benthic mat communities of Lakes Bonney and Fryxell. Viral taxonomic diversity was compared across lakes and ecological function was investigated by characterizing auxiliary metabolic genes (AMGs) and predicting viral hosts. Our data suggest that viral communities differed between the lakes and among sites: these differences were connected to microbial host communities. AMGs were associated with the potential augmentation of multiple biogeochemical processes in host, most notably with phosphorus acquisition, organic nitrogen acquisition, sulfur oxidation, and photosynthesis. Viral genome abundances containing AMGs differed between the lakes and microbial mats, indicating site specialization. Using procrustes analysis, we also identified significant coupling between viral and bacterial communities (p = 0.001). Finally, host predictions indicate viral host preference among the assembled viromes. Collectively, our data show that: (i) viruses are uniquely distributed through the McMurdo Dry Valley lakes, (ii) their AMGs can contribute to overcoming host nutrient limitation and, (iii) viral and bacterial MDV communities are tightly coupled.

Microbiology↗
DOE OSTI2023

Examining the role of glycoside hydrolases in local rheology of Pseudomonas aeruginosa biofilms

Current research strategies in the treatment of biofilm infections have focused on dispersal, in which bacteria are made to vacate the extracellular polymeric substance (EPS) surrounding them and return to a planktonic state where antimicrobial treatments are more effective. Glycoside hydrolases (GHs), which cleave bonds in EPS polysaccharides, have been shown to promote dispersal in Pseudomonas aeruginosa biofilms. The dispersal mechanism is possibly due to GHs’ ability to directly release bacteria from the EPS, disrupt EPS’ ability to regulate the environment, or reduce overall mechanical stability. In this work, passive microrheology is used to examine the relevance of the last mechanism by exploring the effects of three GHs (α-amylase, cellulase, and xylanase) known to disperse P. aeruginosa on local biofilm viscoelasticity. Compared to control studies in wild-type strains, it is found that treatment with all three GHs results in statistically relatively less elastic and stiffer biofilms, indicating that changes to mechanical stability may be a factor in effective dispersal. Both cellulase and xylanase were observed to have the greatest impact in creating a less stiff and elastic biofilm; these GHs have been observed to be effective at dispersal in the published results. Each GH was further tested on biofilms grown with strains that produced EPS missing specific polysaccharide components. Cellulase specifically targeted Psl, which forms the major structural and mechanical backbone of the EPS, explaining its efficacy in dispersal. However, xylanase did not appear to exhibit any affinity to any polysaccharide within the EPS based on the microrheology results. Overall, these results suggest that the local microrheology of the biofilms is impacted by GHs and that may be one of the factors that is causing the ability of these therapeutics to enhance dispersal.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

The Role of Phytoplankton Biomacromolecules in Controlling Ocean Surface Roughness

Satellite altimetric data routinely map sea surface topography by measuring the ocean return signal. One source of altimeter measurement contamination occurs when the radar ocean backscatter becomes unusually large, a situation termed a Sigma-0 bloom. Past research suggests Sigma-0 blooms are associated with weak wind and natural surface slick conditions where capillary waves at the air–sea interface are suppressed. To date, no explicit connection between these conditions and Sigma-0 bloom presence has been provided. Using a series of simplified equations, our reduced model determines capillary wave heights from estimates of planktonic carbon concentrations and regional wind speed. Our results suggest that the radar signal reflection increases as capillary wave height decreases. This relationship depends on surfactant concentration, surfactant composition, and wind speed. Model sensitivity analysis indicates that the interface reflectivity depends on biological activity and wind speed. Our proposed simplified model provides a method to identify potential Sigma-0 bloom regions. We conclude that because of the demonstrated impact of biological surfactants on ocean roughness, it is necessary to consider the biological activity, i.e., phytoplankton bloom events, when interpreting signals from radar altimetry and when developing ocean hydrology models.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2025

Evaluation of cloud height, optical thickness, and phase retrievals from the CHROMA algorithm applied to Sentinel-3 OLCI data

We previously developed the Cloud Height Retrieval from O 2 Molecular Absorption (CHROMA) algorithm for the Ocean Color Instrument (OCI) on the new NASA Plankton, Aerosol, Cloud, ocean Ecosystem (PACE) mission. Here, we apply CHROMA to observations from the Ocean Land Colour Instrument (OLCI) to guide expectations for PACE, as it will take some time to obtain large-scale validation data for OCI. We use cloud top height (CTH), phase, and (for liquid clouds) cloud optical thickness (COT) data from the ground-based Atmospheric Radiation Measurement (ARM) network to evaluate the OLCI retrievals. We found that OLCI and Moderate Resolution Imaging Spectroradiometer (MODIS) CTH compare similarly well to the ARM reference. OLCI has a tendency to underestimate CTH as CTH increases, and algorithm assumptions about cloud geometric thickness may contribute to this. ARM COT from multifilter shadowband radiometers (MFRSR) and Sun photometers are well-correlated with one another, albeit with a roughly 30 % offset on average; OLCI and MODIS COT agree more closely with the MFRSR data. OLCI retrieval uncertainty estimates show skill at telling low-uncertainty cases from high-uncertainty ones, although CTH uncertainties are underestimated. Additionally, we compare the OLCI data to satellite retrievals based on thermal infrared measurements from MODIS and Sea and Land Surface Temperature Radiometer (SLSTR) data. Differences are broadly consistent with physical expectations based on the A-band vs. thermal techniques, although one key challenge in such aggregated comparisons is different cloud masking sensitivities and algorithm failure rates meaning additional sampling differences are introduced. We conclude by discussing the transition to and possible enhancements for PACE OCI.

Sayer, Andrew M. [Univ. of Maryland Baltimore Coun↗
DOE OSTI2021

A bacterial membrane sculpting protein with BAR domain-like activity

Bin/Amphiphysin/RVS (BAR) domain proteins belong to a superfamily of coiled-coil proteins influencing membrane curvature in eukaryotes and are associated with vesicle biogenesis, vesicle-mediated protein trafficking, and intracellular signaling. Here, we report a bacterial protein with BAR domain-like activity, BdpA, from Shewanella oneidensis MR-1, known to produce redox-active membrane vesicles and micrometer-scale outer membrane extensions (OMEs). BdpA is required for uniform size distribution of membrane vesicles and influences scaffolding of OMEs into a consistent diameter and curvature. Cryo-TEM reveals that a strain lacking BdpA produces lobed, disordered OMEs rather than membrane tubules or narrow chains produced by the wild-type strain. Overexpression of BdpA promotes OME formation during planktonic growth of S. oneidensis where they are not typically observed. Heterologous expression results in OME production in Marinobacter atlanticus and Escherichia coli . Based on the ability of BdpA to alter membrane architecture in vivo, we propose that BdpA and its homologs comprise a newly identified class of bacterial BAR domain-like proteins.

60 APPLIED LIFE SCIENCES↗