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At least 235 records · Page 13

Identification and overexpression of endogenous transcription factors to enhance lipid accumulation in the biotechnologically relevant species Chlamydomonas pacifica

Sustainable low-carbon energy solutions are critical to mitigating global carbon emissions. Algae-based platforms offer potential by converting carbon dioxide into valuable products while aiding carbon sequestration. However, scaling algae cultivation faces challenges like contamination in outdoor systems. Previously, our lab evolved Chlamydomonas pacifica, an extremophile green alga, which tolerates high temperature, pH, salinity, and light, making it ideal for large-scale bioproduct production, including biodiesel. Here, we enhanced lipid accumulation in evolved C. pacifica by identifying and overexpressing key endogenous transcription factors through genome-wide in-silico analysis and in-vivo testing. These factors include Lipid Remodeling Regulator 1 (CpaLRL1), Nitrogen Response Regulator 1 (CpaNRR1), Compromised Hydrolysis of Triacylglycerols 7 (CpaCHT7), and Phosphorus Starvation Response 1 (CpaPSR1). Under nitrogen deprivation, CpaLRL1, CpaNRR1, and CpaCHT7 overexpression enhanced lipid accumulation compared to wild-type. However, CpaPSR1 increased lipid accumulation compared to wild-type in normal media and did not increase further under nitrogen deprivation, highlighting the difference in function based on media conditions. Notably, lipid analysis of CpaPSR1 under normal media conditions revealed a 2.4-fold increase in triglycerides (TAGs) compared to the wild-type, highlighting its potential for biodiesel production. This approach provides a framework for transcription factor-focused metabolic engineering in algae, advancing bioenergy and biomaterial production.

Biofuels↗

Scarcity of fixed carbon transfer in a model microbial phototroph–heterotroph interaction

Although the green alga Chlamydomonas reinhardtii has long served as a reference organism, few studies have interrogated its role as a primary producer in microbial interactions. Here, we quantitatively investigated C. reinhardtii’s capacity to support a heterotrophic microbe using the established coculture system with Mesorhizobium japonicum, a vitamin B 12 -producing α-proteobacterium. Using stable isotope probing and nanoscale secondary ion mass spectrometry (nanoSIMS), we tracked the flow of photosynthetic fixed carbon and consequent bacterial biomass synthesis under continuous and diurnal light with single-cell resolution. We found that more 13 C fixed by the alga was taken up by bacterial cells under continuous light, invalidating the hypothesis that the alga’s fermentative degradation of starch reserves during the night would boost M. japonicum heterotrophy. 15 NH 4 assimilation rates and changes in cell size revealed that M. japonicum cells reduced new biomass synthesis in coculture with the alga but continued to divide—a hallmark of nutrient limitation often referred to as reductive division. Despite this sign of starvation, the bacterium still synthesized vitamin B 12 and supported the growth of a B 12 -dependent C. reinhardtii mutant. Finally, we showed that bacterial proliferation could be supported solely by the algal lysis that occurred in coculture, highlighting the role of necromass in carbon cycling. Collectively, these results reveal the scarcity of fixed carbon in this microbial trophic relationship (particularly under environmentally relevant light regimes), demonstrate B 12 exchange even during bacterial starvation, and underscore the importance of quantitative approaches for assessing metabolic coupling in algal–bacterial interactions.

59 BASIC BIOLOGICAL SCIENCES↗

Direct Ink Write and Processing of Complex 3D Marine Compatible Structures with Calcium Carbonate Slurries

Ocean acidification heavily impacts marine ecosystems by reducing calcification. Many coral-algae symbiotic relationships are in jeopardy due to the destruction of coral reefs. Here, a ceramic ink compatible with the direct ink write additive manufacturing technique was formulated and used to print marine compatible structures that could grow algae and restore that relationship. A diacrylate polymer was mixed with calcium carbonate, a material that comprises a coral skeleton, to create a slurry with shear thinning properties. Rheology studies were conducted to confirm printing properties and characterize the slurry. Printed parts demonstrated strong control over print features, including size and infill design. Various infill patterns and percentages were attempted to optimize printability and potential algae growth. Thermogravimetric analysis helped determine a logical burnout and sintering procedure to avoid large cracking. This project developed a printable and sinter-able calcium carbonate ceramic slurry for complex marine-compatible structures and algae growth. This research was conducted in the support of the Eco Reef project.

36 MATERIALS SCIENCE↗

Underlying data for characterization of phycocyanobilin (PCB) biosynthesis in Galdieria sulphuraria

This dataset contains data for studies of phycocyanobilin (PCB) synthesis in the red alga Galdieria sulphuraria. Red algae such as G. sulphuraria utilize phycobilisomes for light harvesting. The phycobilisomes of early-branching organisms such as G. sulphuraria or Cyanidioschyzon merolae contain PCB chromophores but not phycoerythrobilin (PEB), in contrast to the phycobilisomes of other red algae. The studies reported in this dataset examine biosynthesis of PCB in G. sulphuraria and C. merolae, starting from biliverdin IX-alpha (BV), the last known common precursor for PCB and PEB. In cyanobacteria, phages, green algae, and land plants, conversion of BV into PCB or PEB is carried out by a family of enzymes called ferredoxin-dependent bilin reductases (FDBRs). The current studies demonstrate that G. sulphuraria, but not C. merolae, require the action of an additional isomerase to synthesize PCB.

59 BASIC BIOLOGICAL SCIENCES↗

Observations on gas exchange and element recycle within a gas-closed algal-mouse system

Life support systems based on bioregeneration rely on the control and manipulation of organisms. Algae are potentially useful for a variety of Closed Ecological Life Support System (CELSS) functions including the revitalization of atmospheres, production of food and for nitrogen fixation. The results of experiments conducted with a gas-closed algal-mouse system designed to investigate gas exchange phenomena under varying algal environmental conditions, and the ability of algae to utilize oxidized mouse solid waste are reported. Inherent instabilities exist between the uptake and release of carbon dioxide (CO2) and oxygen (O2) by the mouse and algae in a gas-closed system. Variations in light intensity and cell density alter the photosynthetic rate of the algae and enable short-term steady-state concentrations of atmospheric CO2 and O2. Different nitrogen sources (urea and nitrate) result in different algal assimilatory quotients (AQ). Combinations of photosynthetic rate and AQ ratio manipulations were examined for their potential in stabilizing atmospheric gas concentrations in the gas-closed algal-mouse system.

Smernoff, D. T.↗

Algal Accessory Pigment Detection Using AVIRIS Image-Derived Spectral Radiance Data

Visual and derivative analyses of AVIRIS spectral data can be used to detect algal accessory pigments in aquatic communities. This capability extends the use of remote sensing for the study of aquatic ecosystems by allowing detection of taxonomically significant pigment signatures which yield information about the type of algae present. Such information allows remote sensing-based assessment of aquatic ecosystem health, as in the detection of nuisance blooms of cyanobacteria or toxic blooms of dinoflagellates. Remote sensing of aquatic systems has traditionally focused on quantification of chlorophyll a, a photoreactive (and light-harvesting) pigment which is common to all algae as well as cyanobacteria (bluegreen algae). Due to the ubiquitousness of this pigment within algae, chl a is routinely measured to estimate algal biomass both during ground-truthing and using various airborne or satellite based sensors, including AVIRIS. Within the remote sensing and aquatic sciences communities, ongoing research has been performed to detect algal accessory pigments for assessment of algal population composition. This research is based on the fact that many algal accessory pigments are taxonomically significant, and all are spectrally unique. Aquatic scientists have been refining pigment analysis techniques, primarily high performance liquid chromatography, or HPLC, to detect specific pigments as a time-saving alternative to individual algal cell identifications and counts. Remote sensing scientists are investigating the use of pigment signatures to construct pigment libraries analogous to mineral spectral libraries used in geological remote sensing applications. The accessory pigment approach has been used successfully in remote sensing using data from the Thematic Mapper, low-altitude, multiple channel scanners, field spectroradiometers and the AVIRIS hyperspectral scanner. Due to spectral and spatial resolution capabilities, AVIRIS is the sensor of choice for such studies. We present here our results on detection of algal accessory pigments using AVIRIS data.

Richardson, Laurie L.↗

Method of producing purified carotenoid compounds

A method of producing a carotenoid in solid form includes culturing a strain of Chlorophyta algae cells in a minimal inorganic medium and separating the algae comprising a solid form of carotenoid. In one embodiment f the invention, the strain of Chlorophyta algae cells includes a strain f Chlamydomonas algae cells.

Eggink, Laura↗

Quantifying Spatiotemporal Variability of Glacier Algal Blooms and the Impact on Surface Albedo in Southwestern Greenland

Albedo reduction due to light-absorbing impurities can substantially enhance ice sheet surface melt by increasing surface absorption of solar energy. Glacier algae have been suggested to play a critical role in darkening the ablation zone in southwestern Greenland. It was very recently found that the Sentinel-3 Ocean and Land Colour Instrument (OLCI) band ratio R709 nm∕R673 nm can characterize the spatial patterns of glacier algal blooms. However, Sentinel-3 was launched in 2016, and current data are only available over three melting seasons (2016–2019). Here, we demonstrate the capability of the MEdium Resolution Imaging Spectrometer (MERIS) for mapping glacier algae from space and extend the quantification of glacier algal blooms over southwestern Greenland back to the period 2004–2011. Several band ratio indices (MERIS chlorophyll a indices and the impurity index) were computed and compared with each other. The results indicate that the MERIS two-band ratio index (2BDA) R709 nm∕R665 nm is very effective in capturing the spatial distribution and temporal dynamics of glacier algal growth on bare ice in July and August. We analyzed the interannual (2004–2011) and summer (July–August) trends of algal distribution and found significant seasonal and interannual increases in glacier algae close to the Jakobshavn Isbrae Glacier and along the middle dark zone between the altitudes of 1200 and 1400 m. Using broadband albedo data from the Moderate Resolution Imaging Spectroradiometer (MODIS), we quantified the impact of glacier algal growth on bare ice albedo, finding a significant correlation between algal development and albedo reduction over algae-abundant areas. Our analysis indicates the strong potential for the satellite algal index to be used to reduce bare ice albedo biases in regional climate model simulations.

surface albedo↗

Multicriteria-Based Selection of Microalgae Biorefineries: Biomass Composition Defines the Most Suitable Product Portfolios

The supply of microalgae-derived biofuels and bioproducts will be vital in a global-scale bioeconomy. As the diversity of microalgae strains and their compositional plasticity may yield a wide array of products of market interest, the design of effective biorefineries is a central aspect in the path to making microalgae-based products available in the market. In this way, the conversion of microalgae biomass should be planned to employ mature technologies, while maximizing economic and environmental benefits obtained from a diverse product portfolio. This study applied sequential, hybrid Multicriteria Decision Analyses (MCDA) to aid the decision-making process of outlining the most suitable biorefining pathways for specific compositional profiles. For this, the methodology simultaneously considered multiple technical, economic, and environmental criteria, such as market aspects for the main algae-derived products, potential reduction in greenhouse gas (GHG) emissions provided by the biobased alternatives, and technology readiness level of conversion routes, among others. The framework was tested using productivity and compositional data for 13 high-productivity summer strains cultivated under varying nutrient availability. The analysis pointed to compositional profile being a key driver in defining the core biorefining strategy of algae biomass, with lipid-to-carbohydrate ratios higher than roughly 1 warranting the preferential processing of lipids into hydrocarbon fuels with the remainder of the algae biomass compounds being sent to higher-value applications, such as carboxylic acids and renewable thermoplastic substitutes. An in-depth process simulation, techno-economic assessment (TEA), and life-cycle analysis (LCA) effort was carried out as a closing step to corroborate the results stemming from the proposed framework. This study validates the use of MCDAs as a screening method prior to implementing more time-consuming analysis techniques and makes a compelling case for this approach as a product selection tool on a wide range of algae species, thus helping establish species-agnostic (but composition-driven) biorefineries.

biofuels↗

Scale-up study of enhancing algal growth through bioaugmentation with the indole-3-acetic acid producing bacteria Azospirillum brasilense

Algae-based wastewater treatment technologies can recover nutrients while generating algal biomass with diverse application potentials. While numerous bacteria have demonstrated the ability to enhance algal growth through symbiotic relationships in laboratory settings, scaling this benefit to larger applications remains challenging due to complex wastewater conditions and microbe-algae dynamics. This study explores the effectiveness of bioaugmentation with Azospirillum brasilense, a plant growth-promoting bacterium known for producing the phytohormone indole-3-acetic acid (IAA), in boosting algal productivity within an Algaewheel wastewater treatment system. Through a series of experiments conducted at lab-, pilot-, and full-scale levels, we evaluated the impact of bioaugmentation on algal growth and developed a bioaugmentation strategy for an Algaewheel system. The results indicate that a biweekly dosing, achieving a final bioaugmenting cell density of 0.2–4 × 10 9 cells/L, effectively doubles the biomass productivity in a residential subdivision wastewater treatment plant in Northern Illinois. The bacterial production of IAA was the primary mechanism driving this enhancement, as demonstrated by the similar growth and yield improvements observed with chemically supplemented IAA and bioaugmentation across all experimental scales. High-throughput sequencing of 18S rRNA genes revealed that the algal community in the bioaugmented tank exhibited more stable biodiversity than the control. Additionally, bioaugmentation improved nutrient removal efficiency during winter, highlighting its potential to enhance the overall performance and sustainability of algae-based wastewater systems. The developed bioaugmentation and chemical treatment methods offer further operational solutions for managing yield, optimizing biochemical profiles, and enhancing biofuel production potential.

Algae-bacterial symbiotic relationship↗

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↗

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

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

09 BIOMASS FUELS↗

Unraveling metabolism underpinning biomass composition shift in Scenedesmus obliquus under simulated outdoor conditions using 13 C-fluxomics

To render the resulting biomass more attractive and amenable for utilization as the basis for low-carbon intensity bioproducts, single-celled algae need to be biochemically and metabolically poised to assimilate and store the delivered carbon in the fastest and most efficient manner. Accelerating biochemical carbon storage, as primarily carbohydrates or lipids, is critical to achieve the high carbon capture potential that is assigned to algae. To guide strain optimization and engineering for maximizing carbon capture and storage, it is essential to elucidate the link between carbon metabolism and biomass composition. Most published metabolomics work in algae remains largely restricted to ideal and simplified environmental conditions in model organisms, thereby limiting their translation to outdoor implementation. In this work, we utilize 13 C isotopic labeling to characterize distinct intracellular metabolic fluxes before, during, and after nitrogen depletion-induced compositional shifts in Scenedesmus obliquus UTEX 393. The results indicate that a transition to carbohydrates is characterized by diverting flux to starch instead of replenishing the Calvin cycle for CO 2 fixation whereas the subsequent transition to lipids is fueled by NADPH produced by upregulating the phosphoenolpyruvate carboxylase (PEPC)–malic enzyme (ME) cycle flux. Our work highlights bottlenecks to carbohydrate- and lipid-rich biomass and can guide implementable strategies to control the fate of fixed carbon in S. obliquus.

09 BIOMASS FUELS↗

Probable calcified metaphytes in the latest Proterozoic Nama Group, Namibia: origin, diagenesis, and implications

Samples from the Huns Limestone Member, Urusis Formation, Nama Group, at two adjacent localities in southern Namibia contain thin foliose to arched, sheet-like carbonate crusts that are 100-500 micrometers thick and up to 5 cm in lateral dimension. Morphologic, petrographic, and geochemical evidence supports the interpretation of these delicate crusts as biogenic, most likely the remains of calcified encrusting metaphytes. The original sediments of the fossiliferous samples contained aragonitic encrusting algae, botryoidal aragonite cements, and an aragonite mud groundmass. Spherulites within the precursor mud could represent bacterially induced mineral growths or the concretions of marine rivularian cyanobacteria. Original textures were severely disrupted during the diagenetic transition of aragonite to low-magnesian calcite, but some primary structures remain discernible as ghosts in the neomorphic mosaic. Gross morphology, original aragonite mineralogy, and hypobasal calcification indicate that the crusts are similar to late Paleozoic phylloid algae and extant peyssonnelid red algae. Structures interpreted as possible conceptacles also suggest possible affinities with the Corallinaceae. Two species of Cloudina, interpreted as the remains of a shelly metazoan, are also known from limestones in the Nama Group. It is possible, therefore, that skeletalization in metaphytes and animals arose nearly simultaneously near the end of the Proterozoic Eon.

NASA Discipline Number 52-30↗

Integration of CO 2 capture and microalgae cultivation: demonstration assessment of outdoor Scenedesmus acutus cultivation using gaseous ammonia as a nitrogen source

Towards the goal of integrating CO 2 capture using aqueous ammonia with its utilization for algae cultivation, Scenedesmus acutus (UTEX B72) was grown in 1100 L open raceway ponds using CO 2 and NH 3 supplied from gas cylinders. CO 2 /NH 3 mole ratios of 7 and 10 were employed, the gas mixture acting as a surrogate for the output from a CO 2 scrubbing system using aqueous ammonia. Compared to Scenedesmus acutus grown in open ponds using gaseous CO 2 and NaNO 3 as the N-source, the ponds supplied with gaseous CO 2 and NH 3 displayed higher productivity at both CO 2 /NH 3 ratios, with the higher ratio providing the best growth. Depending on the culturing conditions and CO 2 /NH 3 ratio, CO 2 utilization ranged up to 15.8% and NH 3 utilization to 23.0%. These rather low values reflect the fact the high CO 2 /NH 3 feed rate used, resulting in a substantial release of NH 3 from the ORPs (~45%). Finally, these findings demonstrate the suitability of gaseous NH 3 as a N-source for microalgae cultivation, while highlighting the need for a control strategy that closely balances the CO 2 /NH 3 supply with the algae growth rate. The produced algae biomass possessed a high protein and low ash content, rendering it particularly suitable for use as a bioplastic feedstock.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

An Algal Nutrient‐Replete, Optimized Medium for Fast Growth and High Triacylglycerol Accumulation

Microalgae are promising sources to sustainably meet the global needs for energy and products. Algae grow under different trophic conditions, where nutritional status regulates biosynthetic pathways, energy production, and growth. The green alga Chromochloris zofingiensis has strong economic potential because it co-produces biofuel precursors and the high-value antioxidant astaxanthin while accumulating biomass when grown mixotrophically. As an emerging reference alga for photosynthesis, metabolism, and bioproduction, C. zofingiensis needs a defined, optimized medium to standardize experiments during fast growth for batch cultivation. Because the interplay of glucose treatment (+Glc) and mineral deficiency influences photosynthesis, growth, and the production of lipids and astaxanthin, we designed a replete nutrient medium tailored to the C. zofingiensis cellular ionome. We combined inductively coupled plasma mass spectrometry (ICP-MS) and +Glc growth curves to determine a medium that is nutrient replete for at least 5 days of +Glc logarithmic growth. We found that there are high nutritional needs for phosphorus and sulfur during mixotrophy. Iron was the only element measured for which the cellular concentration correlated with exogenous concentration and was iteratively adjusted until the cellular ionome was consistent through the logarithmic growth phase. This Chromochloris-Optimized Ratio of Elements (CORE) medium supports fast growth and high biomass and lipid accumulation without causing excess nutrient toxicity. This defined, nutrient-replete standard is important for future C. zofingiensis investigations and can be adapted for other species to support high biomass for batch cultivation. The method used to develop CORE medium shows how ionomics informs replicable media design and may be applied in industrial settings to inform cost-effective biofuel production.

Jeffers, Tim L. [University of California, Berkele↗

Salinity Stress and the Holobiont: Investigating Algal-Bacterial Interactions in Chrysochromulina tobinii

Algae and bacteria form a symbiotic partnership, known as a ”holobiont”, in which both partners provide critical nutrients to each other. Previous research indicates that the composition and structure of the holobiont plays a crucial role in the algal response to environmental stressors, with expression trends varying under stress conditions. Given the increasing prevalence of salinity shifts due to climate change, investigating the responses of these aquatic organisms to salt stress is essential. This is especially true because algae are ecologically and economically important, serving as a nutritive food source for a large range of eco-cohorts and holding potential for biofuel and nutraceutical applications. This study aimed to examine how holobiont community structure is altered in response to salinity shifts and to assess the corresponding changes in gene expression. Metagenomic and metatranscriptomic approaches were utilized to analyze the interactions between the algae, Chrysochromulina tobinii, and its associated bacterial community under varying salinity conditions. Our findings provide insight into the metabolic adaptations of the holobiont and enhance understanding of the survival mechanisms employed in response to osmotic stress.

59 BASIC BIOLOGICAL SCIENCES↗