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At least 37 records · Page 2

Marine algae growth and carbon capture augmentation by antioxidant nanozymes

Large scale commercial cultivation of microalgae year-round is limited by seasonal stress conditions. The rapid growth and high CO 2 capture of the marine microalga Picochlorum celeri is largely inhibited under winter stress conditions of low temperature and high light. Herein, we demonstrated a nanotechnology approach to enhance the biomass productivity and CO 2 capture of P. celeri under abiotic stress by interfacing with antioxidant cerium oxide nanozymes (nanoceria). Antioxidant nanoceria catalytically scavenged reactive oxygen species (ROS) generated under stress conditions, reducing damage to the microalgae photosynthetic machinery in chloroplasts. Negatively charged poly-acrylic acid-coated nanoceria (PNC, 10 μM) were biocompatible in microalgae cells and colocalized with chloroplasts. In contrast, positively charged aminated nanoceria (ANC) resulted in microalgae aggregation (>50 μM) and were toxic at all concentrations tested (≥10 μM). PNC reduction of ROS levels in microalgae (78%) and superoxide levels (26%), enhanced microalgae growth (65%), photosynthetic performance (130%), and CO 2 uptake rate (380%) under low-temperature stress (15 °C) and high light (500 μmol/m 2 /s PAR) stress relative to controls without nanoceria. Nanoceria augmentation of microalgae provides a rapid and facile technology to increase algae CO 2 capture and biomass under stress conditions.

09 BIOMASS FUELS

Investigating overflow metabolism in heterotrophic cultures of the green alga Chromochloris zofingiensis

Chromochloris zofingiensis is of interest for its ability to perform a reversible trophic switch in the presence of glucose that is characterized by a shutdown of photosynthesis and an accumulation of energy storage metabolites. Previous work has shown that this trophic switch is accompanied by overflow metabolism and the production of lactate in aerobic conditions. This trophic switch is not observed in nutrient replete media. We utilized isotopically assisted metabolic flux analysis to characterize intracellular flux distributions that are associated with different metabolic phenotypes observed in this organism in different media formulations in light and dark conditions. The results of this analysis showed that low iron cultures have no flux through carbon fixation reactions, and that the carbon flux entering the TCA cycle in these cultures is approximately 40 % lower than that in iron replete cultures grown heterotrophically. This analysis was complemented with transcriptomics data collected for C. zofingiensis grown in iron limited conditions to provide further evidence towards the negative impact of iron limitation on both photosynthetic and respiratory activity. Overflow metabolism allows this alga to compensate for the lower energy production that results from iron limitation. This work highlights how nutrient availability can lead to changes in the metabolism of C. zofingiensis.

59 BASIC BIOLOGICAL SCIENCES

Remediation and upcycling of microplastics by algae with wastewater nutrient removal and bioproduction potential

Microplastics have emerged as major environmental hazards that require efficient, cost-effective, and sustainable remediation technologies. This study introduces an integrative platform for the remediation and upcycling of microplastics by algae, while synergizing with plastic upcycling, wastewater treatment, and algal production. The strategy employs a mechanism that enhances hydrophobic interactions between the cell surface and microplastics, enabling rapid aggregation and removal. The platform achieves a superior microplastic removal efficiency of 91.4% within 1 hour, with a capacity of 0.1-gram microplastic per gram of biomass. Furthermore, the study demonstrates an upcycling strategy that converts microplastics-enriched cyanobacteria into plastic composites with unique performance. This work also integrates microplastic removal with cyanobacterial bioproduction and wastewater treatment, offering an approach that synergizes remediation with these value-added processes. Ultimately, this platform provides a viable and sustainable pathway to address microplastic pollution by creating value through plastic upcycling, wastewater nutrient removal, and CO 2 -based bioproduction.

09 BIOMASS FUELS

AlgaeOrtho, a bioinformatics tool for processing ortholog inference results in algae

Introduction: Microalgae constitute a prominent feedstock for producing biofuels and biochemicals by virtue of their prolific reproduction, high bioproduct accumulation, and the ability to grow in brackish and saline water. However, naturally occurring wild type algal strains are rarely optimal for industrial use; therefore, bioengineering of algae is necessary to generate superior performing strains that can address production challenges in industrial settings, particularly the bioenergy and bioproduct sectors. One of the crucial steps in this process is deciding on a bioengineering target: namely, which gene/protein to differentially express. These targets are often orthologs which are defined as genes/proteins originating from a common ancestor in divergent species. Although bioinformatics tools for the identification of protein orthologs already exist, processing the output from such tools is nontrivial, especially for a researcher with little or no bioinformatics experience. Methods: The present study introduces AlgaeOrtho, a user-friendly tool that builds upon the SonicParanoid orthology inference tool (based on an algorithm that identifies potential protein orthologs based on amino acid sequences) and the PhycoCosm database from JGI (Joint Genome Institute) to help researchers identify orthologs of their proteins of interest in multiple diverse algal species. Results: The output of this application includes a table of the putative orthologs of their protein of interest, a heatmap showing sequence similarity (%), and an unrooted tree of the putative protein orthologs. Notably, the tool would be instrumental in identifying novel bioengineering targets in different algal strains, including targets in not-fully annotated algal species, since it does not depend on existing protein annotations. We tested AlgaeOrtho using three case studies, for which orthologs of proteins relevant to bioengineering targets, were identified from diverse algal species, demonstrating its ease of use and utility for bioengineering researchers. Discussion: This tool is unique in the protein ortholog identification space as it can visualize putative orthologs, as desired by the user, across several algal species.

09 BIOMASS FUELS

Genetically pliable green algae for bioproduction of modified fatty acids, nutritional therapeutic oils, and biopharmaceuticals

Homologous recombination (HR) is an essential tool for complex metabolic engineering in yeast, but transgene integration into plant and green algal nuclear genomes predominantly occurs by non-homologous end-joining. Species of the closely related, oleaginous trebouxiophytes Auxenochlorella and Prototheca, are unusual among the green algae in that HR is the favored mechanism for DNA integration into the nuclear genome. This property enables locus-specific targeting of gene cassettes encoding multiple enzymes for manipulating existing biochemical pathways or introducing new functions. Genetic malleability, and regulatory approval for human consumption, coupled with robust fermentation performance at industrial scale, establishes Auxenochlorella and Prototheca as prime candidates for algal production of biochemicals and biomaterials. The examples presented here highlight strain improvement and engineering for synthesis of hydroxylated fatty acids for biomaterials, structured triglycerides resembling human milk fat for infant nutrition, very-long-chain mono- and polyunsaturated fatty acids with nutraceutical or therapeutic potential, and cannabinoids for pharmacological applications.

Moseley, Jeffrey L. [University of California, Ber

Structural Conservation of the A 1 Binding Site in Photosystem I across Cyanobacteria and Green Algae

Time-resolved step-scan Fourier transform infrared (FTIR) difference spectroscopy was used to obtain (A 1 − − A 1 ) FTIR difference spectra from photosystem I (PSI) samples isolated from eight phylogenetically diverse cyanobacterial strains and one green alga, totaling 13 PSI preparations. These included samples from cells grown under farred light and PSI in monomeric, dimeric, trimeric, and tetrameric states. Spectral profiles were shown to be independent of oligomeric state. Remarkably, all (A 1 − − A 1 ) FTIR difference spectra exhibited high similarity, underscoring the robustness of the technique and indicating minimal experimental variability. This congruence reveals a highly conserved environment for the phylloquinone cofactor at the A 1 binding site across diverse taxa. Conserved bands associated with the A 0 pigment further suggest structural continuity from A 0 to A 1 . To leverage this consistency, we constructed a composite (A 1 − − A 1 ) FTIR difference spectrum by averaging all 13 spectra. This composite spectrum provides enhanced resolution, enabling unambiguous identification of previously unresolved bands. The fact that a highly resolved composite spectrum can be obtained by averaging demonstrates the similarity in the spectra from the different types of samples. Band assignments were refined using prior studies, yielding an improved spectral framework for future investigations of PSI electron transfer cofactors.

Charge transfer

Phototropin connects blue light perception to starch metabolism in green algae

Abstract In photosynthetic organisms, light acts as an environmental signal to control their development and physiology, as well as energy source to drive the conversion of CO 2 into carbohydrates used for growth or storage. The main storage carbohydrate in green algae is starch, which accumulates during the day and is broken down at night to meet cellular energy demands. The signaling role of light quality in the regulation of starch accumulation remains unexplored. Here, we identify PHOTOTROPIN-MEDIATED SIGNALING KINASE 1 (PMSK1) as a key regulator of starch metabolism inChlamydomonas reinhardtii. In its phosphorylated form (PMSK1-P), it activates GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (GAP1), promoting starch biosynthesis. We show that blue light, perceived by PHOTOTROPIN, induces PMSK1 dephosphorylation that in turn represses GAP1 mRNA levels and reduces starch accumulation. These findings reveal a previously uncharacterized blue light-mediated signaling pathway that advances our understanding of photoreceptor-controlled carbon metabolism in microalgae.

Science & Technology - Other Topics

Efficient secretion of a plastic degrading enzyme from the green algae Chlamydomonas reinhardtii

Abstract Plastic pollution has become a global crisis, with microplastics contaminating every environment on the planet, including our food, water, and even our bodies. In response, there is a growing interest in developing plastics that biodegrade naturally, thus avoiding the creation of persistent microplastics. As a mechanism to increase the rate of polyester plastic degradation, we examined the potential of using the green microalgaChlamydomonas reinhardtiifor the expression and secretion of PHL7, an enzyme that breaks down post-consumer polyethylene terephthalate (PET) plastics. We engineeredC. reinhardtiito secrete active PHL7 enzyme and selected strains showing robust expression, by using agar plates containing a polyester polyurethane (PU) dispersion as an efficient screening tool. This method demonstrated the enzyme’s efficacy in degrading ester bond-containing plastics, such as PET and bio-based polyurethanes, and highlights the potential for microalgae to be implemented in environmental biotechnology. The effectiveness of algal-expressed PHL7 in degrading plastics was shown by incubating PET with the supernatant from engineered strains, resulting in substantial plastic degradation, confirmed by mass spectrometry analysis of terephthalic acid formation from PET. Our findings demonstrate the feasibility of polyester plastic recycling using microalgae to produce plastic-degrading enzymes. This eco-friendly approach can support global efforts toward eliminating plastic in our environment, and aligns with the pursuit of low-carbon materials, as these engineered algae can also produce plastic monomer precursors. Finally, this data demonstratesC. reinhardtiicapabilities for recombinant enzyme production and secretion, offering a “green” alternative to traditional industrial enzyme production methods.

Science & Technology - Other Topics

Targeted genetic manipulation and yeast-like evolutionary genomics in the green alga Auxenochlorella

Auxenochlorella spp. are diploid oleaginous green algae whose streamlined genomes can be readily manipulated by homologous recombination, making them highly amenable to discovery research and bioengineering. Vegetatively diploid organisms experience specific evolutionary phenomena, including allodiploid hybridization, mitotic recombination, loss-of-heterozygosity, and aneuploidy; however, studies of these forces have largely focused on yeasts. Here, we present a telomere-to-telomere phased diploid genome assembly of Auxenochlorella UTEX 250-A (haploid length 22 Mb) and introduce a genetic toolkit for site-specific manipulation of the nuclear genome in multiple strains, featuring several selectable markers, inducible promoters, and fluorescent reporters for protein localization. UTEX 250-A is an allodiploid hybrid of Auxenochlorella protothecoides and Auxenochlorella symbiontica, two species differentiated by extensive chromosomal rearrangements. UTEX 250-A haplotypes are a mosaic of each parental species following mitotic recombination, and two chromosomes are trisomic. Loss-of-heterozygosity events are pervasive across Auxenochlorella and can evolve rapidly in the laboratory. High-quality structural annotation yielded ∼7,500 genes per haplotype. Auxenochlorella have experienced gene family loss and reduction, including core photosynthesis genes, and exhibit periodic adenine and cytosine methylation at promoters and gene bodies, respectively. Approximately 10% of genes, especially those involved in DNA repair and sex, overlap antisense long noncoding RNAs, which may participate in a regulatory mechanism. We demonstrate the utility of Auxenochlorella for fundamental research by knockout of a chlorophyll biosynthesis enzyme, and confirm one trisomy by allele-specific transformation. These results demonstrate the generality of several evolutionary forces associated with vegetative diploidy and provide a foundation for the use of Auxenochlorella as a reference organism.

CHL27

Integrating an Industrial Source and Commercial Algae Farm with Innovative CO 2 Transfer Membrane and Improved Strain Technologies

This report describes the overall findings of the research project “Integrating an Industrial Source and Commercial Algae Farm with Innovative CO 2 Transfer Membrane and Improved Strain Technologies”. The motivation and goal for this project were to increase the carbon utilization efficiency (CUE) and areal productivity for algal cultivations, thereby reducing CO 2 costs to cultivation operations and improving economics. This was achieved through a combination of enhanced delivery of inorganic carbon and improved strains of Nannochloropsis oceanica capable of higher rates of bicarbonate uptake and metabolism. The project succeeded in these goals. First, a bubble-free, membrane-based technology was developed for delivering CO 2 to cultivations, which increased the CUE from the 15-20% that is standard in the industry to more than 65%. In addition, N. oceanica was modified to express a bicarbonate transporter protein, BicA, which enabled the cells to grow more quickly. In addition, advances were made in the protein engineering of carbonic anhydrase (CA) for enhanced stability and catalytic performance, the computational fluid dynamics modeling of algal cultivation systems, and in the life-cycle assessment and technoeconomic analysis of algal production. Together, these outcomes contribute to advancing algal cultivation as an economically viable platform for production of fuels, materials, and other chemical products. The project results aid in addressing the dual challenges of reducing atmospheric CO 2 levels and achieving green energy solutions.

09 BIOMASS FUELS

Evaluation of DNA Extraction Efficiency in Diverse Algae Strains Using Commercial Kits and Lysis Approaches

Efficient DNA extraction is essential for accurately monitoring microalgae communities in large-scale cultivation systems such as raceway ponds and wastewater ponds. Traditional phenol chloroform extracts are a staple in microbiology but are obsolete for routine sampling due to its high toxicity reagents and time intensive setups. Commercial DNA extraction kits are more favorable for the microbes found in these ponds, but lack specific kits made for these communities. Little is known about which kits perform the best, leading researchers to use a variety of different kits with inconsistent results. This project compared one precipitation based commercial kit (Lucigen Masterpure) and five wash based kits (Monarch, Zymo Quick-DNA, and three Qiagen DNeasy kits) using four brackish algae strains to determine which methods yield the greatest quantity and quality of genomic DNA. Extractions were evaluated using the manufacturers protocol, and additional pretreatment options were administered before a single kit to compare its potential in being added routinely before extractions. Pretreatment options included both cryogenic freeze-thawing and heat incubation using enzymes. DNA was quantified using Qubit fluorometry and NanoDrop purity ratios. Overall, the Qiagen PowerWater kit provided the highest DNA yield and purity, but at a significantly higher cost then the precipitation-based kit (MasterPure). It was also noted that while the precipitation-based kit was significantly cheaper, provided similar results, it took significantly more time to complete a single run. Cryogenic pretreatment (6x cycles) increased average DNA yields by up to 80%, whereas enzymatic pretreatment most improved purity ratios without substantially improving quantity. The results suggest that it may be more cost and time efficient to use Qiagen kits with the addition of lysis pretreatments to procure better results. Future works includes developing a better system to efficiently collect multi variable data, and to upscale to artificial polycultures using similar methodologies alongside sequencing to confirm kit results.

59 BASIC BIOLOGICAL SCIENCES

Solar Spectrum Conversion for an Algae Bioreactor (CRADA Final Report)

This project focused on developing advanced optical coatings to improve solar energy utilization. The research aimed to create lanthanide-doped upconversion nanoparticles (UCNPs) capable of capturing unused near-infrared (NIR) light from the sun and converting it into visible light (blue and red photons) that can be used for photosynthesis. The primary goal was to identify, synthesize, and integrate highly efficient UCNPs into a transparent thin-film device. Through a comprehensive workflow involving computer simulations, high-throughput robotic synthesis, and detailed optical characterization, the project successfully developed a high-performance material. The key technical achievement was the creation of a core-shell UCNP (NaYF₄:20%Yb³⁺, 2%Er³⁺ coated with a 10 nm NaYF₄ shell) that demonstrated a quantum yield of 3.2% for converting 980 nm NIR light into visible light. Transparent thin films fabricated from these nanoparticles showed excellent optical properties, confirming their potential for practical applications. This research adds to the scientific understanding of energy transfer in lanthanide materials and demonstrates a technically effective method for creating efficient light-converting coatings. The primary benefit to the public lies in the potential for these coatings to enhance the efficiency of solar-driven processes, such as boosting the growth of algae in photobioreactors for biofuel production.

14 SOLAR ENERGY

An Experimental and Computational Approach to Investigating CO 2 Uptake of Cellulose-producing Algae from Cellulosic Ethanol Production (Final Report)

This project combined experimental algal cultures with predictive quantum calculations to evaluate system-level CO 2 uptake and conversion efficiency of cellulose-producing Nannochloropsis sp. algae. Recently, Nannochloropsis sp. has garnered attention as a novel host organism for converting low-cost CO 2 produced from cellulosic ethanol fermentations into algal lipids for biodiesel production and microcrystalline cellulose as a high-value co-product. As depicted in the figure below, this project (1) optimized Nannochloropsis salina (N. salina) cultures on effluent gas produced directly from cellulosic ethanol fermentation, (2) characterized the fermentation products, quantify cellulose production, and calculate CO 2 uptake efficiency with predictive quantum calculations, (3) conducted a life cycle and techno economic analysis of the proposed integration, and (4) provided training opportunities to students attending UC Riverside.

09 BIOMASS FUELS

Algae Hydrothermal Liquefaction: End-of-Project Report

This document is a close-out report for two Pacific Northwest National Laboratory (PNNL) projects funded by the Alternative Fuels and Feedstocks Office (formerly the Bioenergy Technologies Office) of the Department of Energy (DOE). The two projects are: • Thermochemical Interface (DOE ref. no. 1.3.4.101, PNNL ref. no. 64490) • HTL Model Development (DOE ref. no. 1.3.5.202, PNNL ref. no. 71612) At the end of fiscal year 2025, both projects ended. Additional activities related to the topics of algae, hydrothermal liquefaction (HTL), and the techno-economic assessment of HTL of algal feedstocks will be studied in other projects. Key technical findings of the projects were published in early 2025 and highlights will be summarized in this document. A summary of recent technical findings from 2025 are also included herein. This close-out report summarizes key outcomes from both projects highlighting the value and technical impact achieved.

09 BIOMASS FUELS

Engineering-Scale Validation of Novel Algae CO 2 Capture and Bioproducts Technology

The purpose of this project was to advance and scale the algae-based technology Helios-NRG has been developing for capture of CO 2 from the flue gas of coal-burning power plants with conversion of the CO 2 to high-value products. In order to encourage the process becoming commercial and be adoptable, it is essential that the cost of CO 2 be close to net zero or slightly revenue positive. Two methods to achieve this are reducing the cost of operations through efficiencies and selling of products derived from conversion. Helios and the project team partners have accomplished the goals of furthering the technology to be more scalable and substantially lower cost with a defined pathway for commercialization.

01 COAL, LIGNITE, AND PEAT

Iron-starvation induces photosystem I antenna remodeling in green algae

Dunaliella salina and Dunaliella tertiolecta are extremophile, marine algae that can survive in very low Fe conditions. In this study, we used TMT-proteomics to compare the Fe starvation responses to the Fe replete responses. Samples were digested with trypsin, labeled with TMT 10-Plex, then analyzed by LC-MS/MS. Data was searched with MS-GF+ using PNNL's DMS Processing pipeline.

59 BASIC BIOLOGICAL SCIENCES