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Whole Algae Hydrothermal Liquefaction and Upgrading: A review of progress and challenges and insight into the future

This report summarizes the research at Pacific Northwest National Laboratory (PNNL) to evaluate the economic viability and environmental impact of using microalgae to produce fuels and other products via hydrothermal liquefaction (HTL). Over the past several years, PNNL has examined key aspects of feedstock cost and availability, formatting and conversion techniques, and the utilization of all HTL products. Investigations of feedstock cost led to opportunities to work with cost-advantaged algal feedstocks that can be provided at minimal cost for HTL processing. Cost-advantaged algae include wastewater-grown algae and harvested algal blooms. Although farm-cultivated algae offer the best possible biomass composition and scalability for HTL processing, the cost of the feedstock is too high to yield an economically competitive biofuel. Processing cost-advantaged feedstocks creates other unique challenges in adapting HTL to upgrade biomass with higher than typical ash content and less preferred composition (low lipid). Despite the challenges, HTL of cost-advantaged algae results in economically competitive pricing scenarios and significant advantages in reducing net emissions below 70% of the petroleum baseline. The utilization of a variety of potential non-fuel products from algal HTL, such as the use of HTL solids as a cement additive, provides a significant reduction in net emissions by offsetting emissions from other carbon-intense products. This report presents an analysis of the research conducted at PNNL to develop an economically and environmentally beneficial process for algae HTL.

09 BIOMASS FUELS↗

Hydrothermal liquefaction of wastewater-grown algae to produce synthetic aviation fuel: A combined experimental study and techno-economic assessment

Large-scale algae farms may someday become a consistent source of biomass feedstock for biofuels. Near-term supplies of algal biomass are available at certain water resource recovery facilities as algae cultivation is used as a method for nutrient recovery from specific effluent streams. Algae grown as a service shifts the value to the service rather than its sole use as a feedstock, which could enable the provision of algal biomass at low to no cost to biofuel producers. Hydrothermal liquefaction (HTL) can readily upgrade wet feedstock slurries, such as algae, to produce a carbon-enriched biocrude. The HTL biocrude can be hydrotreated and distilled, producing a variety of distillate fuels, including synthetic aviation fuel (SAF). We present a pathway, showing the experimental production of SAF from wastewater-grown algae via HTL, along with a techno-economic assessment to identify opportunities for process improvements. Critical quality attributes of the SAF, such as density, viscosity, surface tension, and freeze point, were estimated within the expected fuel experience ranges when compared against petroleum jet fuel. The average minimum fuel selling price of fuels from wastewater-grown algae for breakeven economics was $\$9.04$ per gasoline gallon equivalent (GGE). The sale of co-products such as struvite fertilizers and cement additives can add revenue to reduce the net cost. Ultimately, the selling price is influenced by the scale of the HTL processing facility. Adjusting estimations in the process scale, algae yield, and capital cost estimation can lower the price to $\$6.51$/GGE or raise it to $13.07/GGE.

Biofuels↗

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↗

Balancing Water Sustainability and Productivity Objectives in Microalgae Cultivation: Siting Open Ponds by Considering Seasonal Water-Stress Impact Using AWARE-US

Microalgae have great potential as an energy crop. Scaling-up algal biofuel production in the United States (US) should be done with careful attention to water stress. This study evaluates the regional and seasonal water-stress impact of potential algae-pond deployments in the US. Three site-selection strategies focusing on biomass yield, water-use efficiency (WUE), and water-stress impact, respectively, are applied and compared to meet a US algae biomass production target of 30 million metric tons/yr ash-free dry weight, which converts to 20.8 billion L renewable diesel, via hydrothermal liquefaction. Ranking algae ponds based on biomass yield leads to freshwater consumption of 2.66 km3/yr, resulting in the highest water-stress impact (39.1 US equivalent km3). Under the WUE scenario, water consumption is reduced by 81%, but biomass yield is reduced by 12%. In contrast, adding a water-stress constraint to the biomass-yield ranking reduces water consumption by 50% and water-stress impact by 97%, with a small yield reduction (1.7%). Results show that pond location has a significant effect on water stress and that water stress is not proportional to water consumption or yield. Furthermore, capturing seasonal water patterns is critical for planning because sites in water-abundant regions can have short-term but significant water-stress impacts.

algae, Biofuel, water scarcity footprint, hydrothe↗

Biomolecular Films for Direct Air Capture of CO 2

Efficient utilization of CO 2 is amongst the most critical cost drivers in algal biomass production in open pond systems. CO 2 delivery costs represent approximately 20% of the final biomass selling price in algal mass cultivation systems. Technologies that enable direct air capture (DAC) of atmospheric CO 2 to decouple algae cultivation from CO 2 point sources thus present an opportunity to improve the economics and resource potential of algal biomass. Current DAC technologies typically employ amine- or caustic-based absorption, demanding significant water and/or energy inputs and incurring substantial capital expenditures. Conversely, bio-based approaches to DAC offer a means to bypass conventional technoeconomic and sustainability hurdles. We integrate recent advances in computational metabolic modeling, algal genetic engineering, algal cultivation, and algal biomass upgrading to enable directed localization and self-assembly of carbonic anhydrase molecular films to gas-liquid interfaces for enhanced CO 2 capture and conversion.

09 BIOMASS FUELS↗

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↗

Integrating microbial communities into algal biotechnology: a pathway to enhanced commercialization

Microalgae are increasingly recognized for their potential in wastewater treatment and the sustainable production of feedstock for fuel, feed, food, and other bioproducts. Like conventional agricultural systems, algal cultivation involves complex microbial communities. However, despite their pivotal role in cultivation outcomes, especially at the commodity-scale, the critical interactions between microalgae and their microbiomes are often overlooked. Here we synthesize current knowledge on the taxonomic diversity, ecological roles, and biotechnological potential of algal microbiomes, with a focus on their interactions with algal hosts through nutrient exchange, growth modulation, pathogen defense, and environmental conditioning. We also examine how environmental factors such as nutrient availability, salinity, and temperature influence these interactions. Advances in microbiome engineering, including synthetic biology and ecological approaches, offer opportunities to enhance beneficial algal-microbiome interactions, thereby improving growth, resilience, and yield. These advancements could lead to more sustainable and economically viable microalgae cultivation, with far-reaching implications for environmental management and biotechnological innovation. By addressing key economic and environmental barriers, microbiome engineering holds transformative potential to revolutionize large-scale algae cultivation and provide sustainable solutions to global challenges.

Microbiology↗

FWP FEW0223: "Advanced Manufactured Carbonate Materials for Algal Biomass Production: Joint LLNL SNL Program" (Final Report)

The ability to easily and cheaply transport carbon dioxide (CO 2 ) from coal-fired power plants equipped with capture facilities to multiple, potentially distant utilization sites of widely varying scales will enable wider use of CO 2 captured from flue gas. In the case of algal biomass cultivation, delivery of CO 2 in the form of carbonate-based materials may provide a more efficient and economical method of inorganic carbon delivery than CO 2 sparging, reducing the cost of algae cultivation and increasing the value of carbon capture. Lawrence Livermore National Laboratory (LLNL) has developed advanced manufactured materials that are well-suited for carbon capture, storage, transport, and delivery. These materials consist of aqueous sodium carbonate, which captures CO 2 as sodium bicarbonate, extruded or 3D printed in a CO 2 permeable polymer to increase the surface area and improve capture kinetics. By encapsulating carbonate solutions, LLNL has demonstrated order-of-magnitude enhancements in carbon capture rates compared with liquid carbonates, enabling the use of these (otherwise kinetically-limited) inexpensive, environmentally-benign solvents for carbon capture.

20 FOSSIL-FUELED POWER PLANTS↗

Pilot-Scale Algal Oil Production

The main objective of the project is complete: development of a preliminary planning and design for a pilot-scale algal oil cultivation and processing facility, FEL-3 design with -5% / +15% cost estimate accuracy, and conversion of the algal oil to biofuel in an off-site existing bio-oil refinery. The design basis includes 10 tons per day of dried algae cultivated with CO 2 supplied by direct air capture, electricity supplied solar power, well water supply, zero liquid discharge, off-site extraction, and off-site conversion of oil to biofuel. The design and permitting package is an important milestone in the path to commercialization of algal biofuels and bioproducts as it provides the preliminary design, permitting path, long-term land lease, planning documents, and team needed for success in future engineering, construction, start-up and operations of a pilot-scale farm at a site in Paso Robles, CA. Outcomes of the business assessment include (i) identification of a product spectrum for economical algal biofuels using co-products with markets that are commensurate with production of 6-7 billion gallons per year of sustainable aviation fuel (SAF), renewable diesel, and renewable gasoline, (ii) a path toward near-term contribution of algae oil to SAF, and (iii) an approach for long-term operation of a pilot-scale farm.

09 BIOMASS FUELS↗

Lipids from High-Protein Algae Biomass

Biofuels from renewable and sustainable sources will be a critical component of reducing greenhouse gasses. To this end, in achieving maximum biomass productivity in outdoor ponds at a reasonable cost, algae must be grown at a maximal rate. This current algae cultivation strategy results in high-protein biomass because there is insufficient time for deplete-hold steps that increase lipids but also cost. Thus methods to convert high-protein (>50%) need to be developed for biofuels. The Consolidated Algae Processing (CAP) strategy has previously shown utility on high-carbohydrate algae biomass and has now been expanded to include methods aimed at converting the protein fraction to lipids. After acid-pretreatment, hydrolysate liquor is oxidatively treated to deaminate and convert soluble proteins and amino acids into four main carboxylate acids; formic, acetic, succinic, and propionic. These acids are readily and completely fermented into intracellular lipids using oleaginous yeast. Using a fed-batch fermentation strategy and no added nutrients, we achieved nearly 30% intracellular lipids and 1.3 g/L lipids demonstrating the potential of our expanded CAP process to generate additional lipids for conversion to biofuels as part of an algae-focused biorefinery.

AAD↗

Carbon Capture and Utilization for Protein and Fatty Acids

The unlimited release version of the final report for the "Carbon Capture and Utilization for Protein and Fatty Acids" project. This project advanced an integrated open raceway algae cultivation and processing system to engineering scale for carbon capture and utilization (CCU) from the flue gas of a naphtha-fired power plant.

02 PETROLEUM↗

A validated thermal and biological model for predicting algal productivity in large scale outdoor cultivation systems

Proper assessment of the sustainability of algal products requires an understanding of algal growth rate with regional and temporal resolution. This study developed a bulk growth model that utilizes geospatial data, reactor geometry inputs, and a maximum of six strain-specific parameters to predict algal productivity with temporal and regional resolution. The model requires local weather data to calculate the time-resolved culture temperature which is combined with a biological model based on maximum theoretical carbon fixation to predict the time-resolved algal concentration. Validation of the model against experimental data illustrates an average ac- curacy of 0.27% ± 5.32% when comparing algal concentration at harvest over 45 days of cultivation data for Galdieria sulphuraria 5587.1, and Galdieria sulphuraria Soos grown in photobioreactors (PBRs) in Mesa, AZ (33.4152° N, 111.8315° W). For open raceway ponds (ORPs), an accuracy of 0.9% ± 2.35% was achieved when comparing algal concentration at harvest over 50 days of cultivation data for Chlorella vulgaris, Desmodesmus intermedius, Nannochloropsis oceanica grown in Atlanta, GA (33.7490° N, 84.3880° W) and Mesa, AZ (33.4152° N, 111.8315° W). The validation trials spanned three seasons (winter, spring, and summer) to ensure model ac- curacy throughout the year, and concentration between harvests was modeled with hourly resolution. The validated model is used to assess the productivity at several case-study locations in the United States with Chlorella vulgaris, grown in open raceway ponds and Galdieria sulphuraria grown in vertical flat panel photo- bioreactors. In conclusion, the model has been published as an open-source tool, in an effort to increase the fidelity of future studies that rely on outdoor algae cultivation for feedstock production and allow for time resolved results for future techno-economic and life cycle assessments.

59 BASIC BIOLOGICAL SCIENCES↗

ASU’s DAC polymer-enhanced cyanobacterial bioproductivity (AUDACity)

ASU’s DAC polymer-enhanced cyanobacterial bioproductivity (AUDACity) project aims to demonstrate a novel, scalable method for removing carbon dioxide (CO 2 ) directly from ambient air and delivering it to cyanobacterial cultures to produce commodity biofuel, mid-value protein for supplements, and high value phycocyanin (PC), a natural blue colorant (Figure A). This approach uses low-cost, reusable anion exchange polymers embedded in modular mesh packets, which capture CO 2 during drying cycles when exposed to ambient air, and release concentrated CO 2 into aqueous cultivation systems. The project addresses a critical challenge in energy research needed for developing sustainable, economically viable methods of Direct Air Capture (DAC) that can be integrated with bio-based systems for fuel and chemical production. AUDACity contributes to scientific understanding by integrating materials chemistry, cyanobacterial biology, and system engineering to create a distributed CO 2 delivery platform. Key insights have emerged around the design of biocompatible sorbents, optimization of CO 2 capture-release cycles, and durability of packet-based delivery systems under outdoor conditions. Notably, the team has synthesized and tested a range of polymer sorbents, identified mechanisms of material degradation and fouling, and advanced both lab- and pilot-scale cultivation systems to evaluate performance. From a technical and economic standpoint, AUDACity shows promise for achieving cost-effective CO 2 capture and delivery into aqueous media and biofuel production. Preliminary techno-economic analysis (TEA) indicates that the DAC system based on current performance can reach $\$$680/tonne CO 2 delivered into aqueous solution; with reasonable improvements to sorbent lifetime, sorbent capacity, reducing water uptake the approach could reach $\$$66/tonne by avoiding the need for energy-intensive sorbent regeneration and CO 2 compression, making it more feasible for decentralized deployment. With these costs for CO 2 and by extracting and selling high-value PC ($\$$50/kg) and mid-value protein supplement ($\$$6/kg), the remaining biomass can be hydrothermally treated into biofuel for $\$$2.50/gallon, and would support a small first-of-a-kind biorefinery capable of producing 500 barrels per day of biofuel. The project offers meaningful public benefits by advancing carbon removal technologies that are low-energy, modular, and adaptable to non-arable land and brackish water use. It aligns with national goals to develop advanced biotechnology and supports future pathways for bio-based fuels and products. By enabling direct coupling of CO 2 transfer into aqueous medium and biological carbon utilization, AUDACity lays the groundwork for effective algae cultivation without wasteful CO 2 delivery and is a promising and innovative solution for low-carbon fuel and bioproduct generation contributing to a vigorous bioeconomy.

09 BIOMASS FUELS↗

Life-cycle analysis of microalgae-based polyurethane foams

Polyurethane plastics are essential in many consumer and commercial products such as insulation, furniture, automotive interiors, and clothing. Pathways for producing polyurethane from microalgae offer an opportunity to reduce greenhouse gas emissions and other environmental impacts and can incorporate processes that avoid the use of toxic isocyanates typically used in conventional polyurethane production processes. In this study, the greenhouse gas emissions, fossil energy, and water consumption of biobased polyurethane and biobased non-isocyanate polyurethane were evaluated via life-cycle analysis using the R&D Greenhouse Gases, Regulated Emissions, and Energy Use in Technologies model. Microalgae-based polyurethane foam was found to achieve greenhouse gas emission reductions of up to 79% compared with conventional polyurethane foam production. The greenhouse gas reductions for the non-isocyanate microalgae polyurethane pathway are slightly lower at 58% compared with conventional polyurethane foam. However, it offers additional benefits by reducing toxicity potential compared to the isocyanate polyurethane pathway. The analysis also included a biorefinery-level analysis to evaluate the impact of incorporating polyurethane production into fuel-processing microalgae biorefineries. The sensitivity analyses conducted in this study reveal that improved algae cultivation strategies can lead to decreases of up to 127% and 80% in GHG emissions from the baseline process of Bio-PU and Bio-NIPU, respectively. Likewise, implementation of renewable electricity can result in up to 128% and 74% lower GHG emissions compared to the baseline production of Bio-PU and Bio-NIPU, respectively. Finally, the analysis evaluated different coproduct handling methods including displacement and allocation (based on mass, energy, and market-value). The results suggest that it is important to consider both the displacement and allocation methods as these led to significant differences in the environmental impacts.

36 MATERIALS SCIENCE↗

Continuous measurements of volatile gases as detection of algae crop health

Wide adoption of algae cultivation to produce environmentally sustainable biofuels and fine chemicals is currently hampered by large losses (10 to 30%) incurred by grazer infections. We show the usage of real-time chemical ionization mass spectrometry to rapidly identify gaseous indicators of grazer infections in cyanobacteria cultures. Grazing was detected significantly faster (up to 3 d) using real-time mass spectrometry than the current methods of microscopy and qPCR. By employing this technology, cultivators may be empowered to treat grazer infestations sooner, thereby protecting the crop and enhancing profitability.

09 BIOMASS FUELS↗

Unrealized Critical Lanthanide Extraction from Sea Algae Mining (UNCLE SAM): Domestic production of critical minerals from seawater

The UNCLE-SAM project, under the Biotechnologies to Ensure a Robust Supply of Critical Materials for Clean Energy program, examined the biomining applications of seaweeds for sustainable, domestic production of critical mineral feedstocks. The ocean is a vast reserve of mineralogical wealth including rare earth elements (REEs) and platinum group metal (PGMs). These elements, categorized as “critical minerals”, are used in telecommunication devices, lasers, LED lighting, turbine generators, electric car motors, jet engine alloys, and many other applications. These critical elements are increasingly vital to a thriving, efficient and sustainable society. However, only a few countries in the global market currently produce and export REEs, leading to potential geopolitical supply disruptions. Marine macroalgae, often referred to as seaweeds, bioconcentrate critical minerals from seawater, including REEs and PGMs. Marine algae cultivation can generate a significant amount of biomass with minimal freshwater, fertilizer, and land requirements. In summary, the UNCLE-SAM project successfully evaluated the technological feasibility for marine macroalgal cultivation as a feedstock for critical minerals, explored the biological capacity of different seaweeds to provide economically relevant domestic mineral production, assessed processing techniques for thermal co-conversion of seaweeds into renewable fuel and mineral feedstocks, and executed techno-economic and lifecycle assessments for identifying the most critical gaps in our current understanding to move the technology into commercially relevant deployment. Further development of this technology could transform the bioproduct and REE mining industries and catalyze the development of a more sustainable future.

58 GEOSCIENCES↗

2024 Case Study: Hydrothermal Liquefaction of Biomass Sources at a Wastewater Treatment Facility

A case study was developed for the fiscal year 2024 to examine the potential of wastewater-grown microalgae as a feedstock for hydrothermal liquefaction (HTL) to produce fuels and other products. In this year’s case study, scenarios are investigated in which the algae cultivation process is placed at different points within the water resource recovery facility (WRRF). Additionally, the supply of biomass for HTL is boosted by blending with wastewater solids collected at different locations within the WRRF.

09 BIOMASS FUELS↗

UNrealized Critical Lanthanide Extraction from Sea Algae Mining (UNCLE SAM)

The UNCLE-SAM project, under the Biotechnologies to Ensure a Robust Supply of Critical Materials for Clean Energy program, examined the biomining applications of seaweeds for sustainable, domestic production of critical mineral feedstocks. The ocean is a vast reserve of mineralogical wealth including rare earth elements (REEs) and platinum group metal (PGMs). These elements, categorized as “critical minerals”, are used in telecommunication devices, lasers, LED lighting, turbine generators, electric car motors, jet engine alloys, and many other applications. These critical elements are increasingly vital to a thriving, efficient and sustainable society. However, only a few countries in the global market currently produce and export REEs, leading to potential geopolitical supply disruptions. Marine macroalgae, often referred to as seaweeds, bioconcentrate critical minerals from seawater, including REEs and PGMs. Marine algae cultivation can generate a significant amount of biomass with minimal freshwater, fertilizer, and land requirements. In summary, the UNCLE-SAM project successfully evaluated the technological feasibility for marine macroalgal cultivation as a feedstock for critical minerals, explored the biological capacity of different seaweeds to provide economically relevant domestic mineral production, assessed processing techniques for thermal co-conversion of seaweeds into renewable fuel and mineral feedstocks, and executed techno-economic and lifecycle assessments for identifying the most critical gaps in our current understanding to move the technology into commercially relevant deployment. Further development of this technology could transform the bioproduct and REE mining industries and catalyze the development of a more sustainable future.

09 BIOMASS FUELS↗