Techno-economic uncertainty analysis of wet waste-to-biocrude via hydrothermal liquefaction
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Li, Shuyun↗
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Algae-Based Beneficial Re-use of Carbon Emissions Using a Novel Photobioreactor: a Techno-Economic and Life Cycle Analysis
Despite the many advantages of microalgae, the feasibility of large-scale cultivation requires significant amounts of carbon dioxide (CO 2 ) to enable high growth rates. A synergistic union typically proposed for the supply of CO 2 is the coupling of algal cultivation with emissions from power plants. This study investigates the sustainability of a novel microalgae platform coupled with coal-based flue gas. The proposed system consists of a novel photobioreactor (PBR) for the production of biomass followed by a two-stage dewatering process. Here, a systems model, which quantifies the CO 2 and energy consumption of the proposed system, was developed and the minimum biomass selling price (MBSP) was determined by a techno-economic analysis (TEA). TEA results indicate that a facility with the capacity to capture 30% of the emissions from a 1 MW power plant requires a biomass production of 1280 metric tonnes per year, which when scaled to a nth of kind facility, can produce biomass at a MBSP of $2322 per tonne. The environmental impact of the proposed facility was determined by a life cycle assessment methodology and results indicate a carbon capture potential of 1.16 x 10 4 metric tons of CO 2 equivalent. In addition, an energy analysis indicates a desirable net energy ratio of 0.1, which is lower than conventional PBRs. Discussion focuses on the requirements to reduce biomass production cost, including research investment areas for increasing productivity while decreasing energy requirements.
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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↗
Heterologous expression of phosphite dehydrogenase in the chloroplast or nucleus enables phosphite utilization and genetic selection in Picochlorum spp.
Microalgae present a path to ameliorate problems associated with climate change via capture and reduction of CO2 to sustainable fuels and chemicals. Picochlorum is a genus of algae recently recognized for potential application in these regards due to its high productivity, thermotolerance, and halotolerance. Foundational genetic tools have recently been established in this genus. However, at present, genetic markers are limited, hindering genetic throughput and trait stacking approaches. To expand the suite of genetic tools and markers available for this genus, we sought to heterologously express the phosphite dehydrogenase (ptxD) gene from Pseudomonas stutzeri WM88 in both the nucleus and chloroplast of Picochlorum renovo and Picochlorum celeri. Additionally, the resultant strains allow for utilization of phosphite as a sole phosphorous source and as a nuclear and plastidial selection marker for genetic engineering. Growth analysis indicated comparable growth and composition when transgenic algae were grown in media containing phosphite as a sole phosphorus source, as compared to the conventionally used phosphate. Combined, these results expand the genetic toolbox available to the Picochlorum genus and present a potential crop protection and biocontainment strategy.
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Cost minimization of deoxygenation for control of Vampirovibrio chlorellavorus in Chlorella sorokiniana cultures
Deoxygenation is a strategy for preventing Vampirovibrio chlorellavorus infection of Chlorella sorokiniana cultures. Deoxygenation cost was minimized by sparging with nitrogen gas for only one hour at the onset of night and relying on naturally-occurring dark-respiration to maintain low oxygen concentrations throughout the night. This technique substantially reduced V. chlorellavorus infection in laboratory co-cultures. The cost of the approach was evaluated in outdoor experiments with one pure water and one pathogen-free C. sorokiniana culture, grown in a 150-L sealed, vented, translucent-polyethylene reactor. Unlike the small laboratory reactors, which were also sealed and vented, the much larger outdoor reactor maintained the dissolved oxygen concentration in the C. sorokiniana culture at near zero concentration throughout the night cycle. This demonstrates that large covered tanks in commercial applications would keep oxygen concentration near zero for the entire night. The total nitrogen sparged, per night, per liter of algal culture, was determined for the 150-L reactor, and used to estimate the cost per m2 for large scale raceways. Onsite nitrogen generators are the most cost-effective method to supply a high flow of nitrogen gas to commercial scale raceways. The cost of deoxygenation treatment ranged from $16/ton AFDW algae for a shallow (2 cm) gravity flow system with 75% harvest prior to deoxygenation to over $1,300/ton for a 20 cm depth raceway with plastic tanks.
59 BASIC BIOLOGICAL SCIENCES↗
Agent-based Modeling for the Multi-objective Optimization of Energy Production Pathways: Integrated Techno-Economics and Life Cycle Assessment
This project developed an integrated modeling framework to evaluate how purpose-grown bioenergy feedstocks could support large-scale sustainable aviation fuel production in the United States. Led by Colorado State University with project partners, the work combined geographically resolved analysis, techno-economic assessment, life cycle assessment, multi-objective optimization, and agent-based modeling to examine feedstock performance, deployment potential, land-use implications, and adoption dynamics. The overall goal was to identify practical and sustainable pathways for producing aviation fuel from feedstocks such as energy crops, woody crops, oilseeds, and algae while helping inform policy, investment, and research needs tied to national SAF goals. In essence, the project provides a systems-level assessment of the opportunities and constraints associated with scaling bioenergy-based aviation fuels in the U.S.
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The carbon-concentrating mechanism of the extremophilic red microalga Cyanidioschyzon merolae
Abstract Cyanidioschyzon merolae is an extremophilic red microalga which grows in low-pH, high-temperature environments. The basis of C. merolae ’s environmental resilience is not fully characterized, including whether this alga uses a carbon-concentrating mechanism (CCM). To determine if C. merolae uses a CCM, we measured CO 2 uptake parameters using an open-path infra-red gas analyzer and compared them to values expected in the absence of a CCM. These measurements and analysis indicated that C. merolae had the gas-exchange characteristics of a CCM-operating organism: low CO 2 compensation point, high affinity for external CO 2 , and minimized rubisco oxygenation. The biomass δ 13 C of C. merolae was also consistent with a CCM. The apparent presence of a CCM in C. merolae suggests the use of an unusual mechanism for carbon concentration, as C. merolae is thought to lack a pyrenoid and gas-exchange measurements indicated that C. merolae primarily takes up inorganic carbon as carbon dioxide, rather than bicarbonate. We use homology to known CCM components to propose a model of a pH-gradient-based CCM, and we discuss how this CCM can be further investigated.
60 APPLIED LIFE SCIENCES↗
Community structure and function during periods of high performance and system upset in a full-scale mixed microalgal wastewater resource recovery facility
Microalgae have the potential to exceed current nutrient recovery limits from wastewater, enabling water resource recovery facilities (WRRFs) to achieve increasingly stringent effluent permits. The use of photobioreactors (PBRs) and the separation of hydraulic retention and solids residence time (HRT/SRT) further enables increased biomass in a reduced physical footprint while allowing operational parameters (e.g., SRT) to select for desired functional communities. However, as algal technology transitions to full-scale, there is a need to understand the effect of operational and environmental parameters on complex microbial dynamics among mixotrophic microalgae, bacterial groups, and pests (i.e., grazers and pathogens) and to implement robust process controls for stable long-term performance. Here, we examine a full-scale, intensive WRRF utilizing mixed microalgae for tertiary treatment in the US (EcoRecover, Clearas Water Recovery Inc.) during a nine-month monitoring campaign. We investigated the temporal variations in microbial community structure (18S and 16S rRNA genes), which revealed that stable system performance of the EcoRecover system was marked by a low-diversity microalgal community (D INVSIMPSON = 2.01) dominated by Scenedesmus sp. (MRA = 55 %-80 %) that achieved strict nutrient removal (effluent TP < 0.04 mg·L -1 ) and steady biomass concentration (TSS monthly avg . = 400–700 mg·L −1 ). Operational variables including pH, alkalinity, and influent ammonium (NH 4 + ), correlated positively (p < 0.05, method = Spearman) with algal community during stable performance. Further, the use of these parameters as operational controls along with N/P loading and SRT allowed for system recovery following upset events. Importantly, the presence or absence of bacterial nitrification did not directly impact algal system performance and overall nutrient recovery, but partial nitrification (potentially resulting from NO 2 − accumulation) inhibited algal growth and should be considered during long-term operation. The microalgal communities were also adversely affected by zooplankton grazers (ciliates, rotifers) and fungal parasites (Aphelidium), particularly during periods of upset when algal cultures were experiencing culture turnover or stress conditions (e.g., nitrogen limitation, elevated temperature). Altogether, the active management of system operation in order to maintain healthy algal cultures and high biomass productivity can result in significant periods (>4 months) of stable system performance that achieve robust nutrient recovery, even in winter months in northern latitudes (WI, USA).
32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
Association of uranium with macerals in marine black shales: Insights from the Upper Devonian New Albany Shale, Illinois Basin
We report marine organic-rich shales generally have high uranium content associated with sedimentary organic matter. However, organic matter in black shales is not homogeneous, but instead a mixture of different macerals. The uranium content and organic petrographic composition of 27 New Albany Shale samples indicate differential contributions of macerals to the uranium content in the Upper Devonian New Albany Shale of the Illinois Basin. Specifically, uranium is mainly associated with amorphous organic matter (microbially degraded phytoplankton, zooplankton, and bacterial biomass), where an average uranium content of approximately 550 ppm has been estimated. The amorphous organic matter's lack of structure, as well as admixed with mineral grains (<7.8 μm) suggest that it originated via microbial degradation in surficial sediments. Uranium may have been incorporated into amorphous organic matter during phytoplankton degradation and amorphous kerogen formation early in diagenesis. Pore-water redox conditions and pH control the reduction of uranium (VI) and its incorporation into amorphous organic matter. Alginite derived from Tasmanites algae (average uranium content <10 ppm) contributes minimally to uranium content. Terrestrial organic matter in the New Albany Shale contains on average ~ 20 ppm uranium, and because it accounts for <10% of total organic matter in the New Albany Shale, its contribution is equally negligible.
58 GEOSCIENCES↗
Development of a field-deployable qPCR assay for real-time pest monitoring in algal cultivation systems
Outdoor cultivation is commonly used to produce algal biomass for a variety of bioproducts including food, feed, fuel, pharmaceuticals, and nutraceuticals. Outdoor cultivation ponds are highly susceptible to pest pressures that may lead to periods of low productivity or even entire loss of the algal crop. Consequently, there is a need for rapid, real-time tracking of pests for early intervention to mitigate crop loss. In this work, we describe the development of a field deployable, low-cost qPCR assay for detecting both known and novel pests of a farmed eukaryotic alga species, Nannochloropsis sp. We performed a proximity guided metagenome deconvolution approach (ProxiMeta™) to discover novel pests that temporally correspond to periods of reduced pond productivity. This approach provided high-quality metagenome assemblies that were used to design qPCR probes to detect specific pests of interest. The portable qPCR assay, designed to be deployed at remote field locations, enables low-cost surveillance with a rapid (2 h) turn-around time. Frequent sampling allows for early detection and prompts intervention strategies to remedy infected ponds to minimize crop loss. The qPCR assay was used to successfully detect a known predatory bacterium within the order Bdellovibrionales both in the lab and at a remote field location. Furthermore, we assembled the genome of two novel, site-specific pests in the Saprospiraceae family and successfully designed qPCR probes that differentially detected their presence in two different pond locations. Ultimately, this assay has the potential to monitor multiple pests simultaneously and tailor targets to match likely pest infections that differ across geographical locations, helping to mitigate crop loss on a large scale.
59 BASIC BIOLOGICAL SCIENCES↗
Abiotic and Biotic Damage of Microalgae Generate Different Volatile Organic Compounds (VOCs) for Early Diagnosis of Algal Cultures for Biofuel Production
Open microalgal ponds used in industrial biomass production are susceptible to a number of biotic and abiotic environmental stressors (e.g., grazers, pathogens, pH, temperature, etc.) resulting in pond crashes with high economic costs. Identification of signature chemicals to aid in rapid, non-invasive, and accurate identification of the stressors would facilitate targeted and effective treatment to save the algal crop from a catastrophic crash. Specifically, we were interested in identifying volatile organic compounds (VOCs) that can be used to as an early diagnostic for algal crop damage. Cultures of Microchloropsis gaditana were subjected to two forms of algal crop damage: (1) active grazing by the marine rotifer, Brachionus plicatilis, or (2) repeated freeze–thaw cycles. VOCs emitted above the headspace of these algal cultures were collected using fieldable solid phase microextraction (SPME) fibers. An untargeted analysis and identification of VOCs was conducted using gas chromatography-mass spectrometry (GC-MS). Diagnostic VOCs unique to each algal crop damage mechanism were identified. Active rotifer grazing of M. gaditana was characterized by the appearance of carotenoid degradation products, including β-cyclocitral and various alkenes. Freeze–thaw algae produced a different set of VOCs, including palmitoleic acid. Both rotifer grazing and freeze–thawed algae produced β-ionone as a VOC, possibly suggesting a common stress-induced cellular mechanism. Importantly, these identified VOCs were all absent from healthy algal cultures of M. gaditana. Early detection of biotic or abiotic environmental stressors will facilitate early diagnosis and application of targeted treatments to prevent algal pond crashes. Thus, our work further supports the use of VOCs for monitoring the health of algal ponds to ultimately enhance algal crop yields for production of biofuel.
09 BIOMASS FUELS↗
Tolerance to allelopathic inhibition by free fatty acids in five biofuel candidate microalgae strains
Contaminating organisms (grazers, pathogens, competitors) and self-inhibition by algae-produced allelopathic chemicals are two issues that may limit the productivity of algal cultivation for bioproducts. One potential solution is to identify algal strains that are not affected by allelopathic inhibition even while undesirable organisms are suppressed. Here we used two experiments to test how sensitivity to allelopathy varies across algae. In the first experiment, we tested the sensitivity of five biofuel candidate green algae strains to two allelopathic compounds (i.e., free fatty acids) and found that the degree of inhibition depends strongly on both the species and specific compound. In the second experiment, we exposed one alga (Chlorella) to the sterile-filtered medium of each species, and found that the concentration of free fatty acids released into the media predicted Chlorella’s growth response. This provides a better understanding of how the production of, and sensitivity to, allelopathic compounds determines algal productivity.
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Identification of Effector Metabolites Using Exometabolite Profiling of Diverse Microalgae
Dissolved exometabolites mediate algal interactions in aquatic ecosystems, but microalgal exometabolomes remain understudied. We conducted an untargeted metabolomic analysis of nonpolar exometabolites exuded from four phylogenetically and ecologically diverse eukaryotic microalgal strains grown in the laboratory, freshwater Chlamydomonas reinhardtii, brackish Desmodesmus sp., marine Phaeodactylum tricornutum, and marine Microchloropsis salina, to identify released metabolites based on relative enrichment in the exometabolomes compared to cell pellet metabolomes. Exudates from the different taxa were distinct, but we did not observe clear phylogenetic patterns. We used feature-based molecular networking to explore the identities of these metabolites, revealing several distinct di- and tripeptides secreted by each of the algae, lumichrome, a compound that is known to be involved in plant growth and bacterial quorum sensing, and novel prostaglandin-like compounds. We further investigated the impacts of exogenous additions of eight compounds selected based on exometabolome enrichment on algal growth. Of these compounds, five (lumichrome, 5'-S-methyl-5'-thioadenosine, 17-phenyl trinor prostaglandin A2, dodecanedioic acid, and aleuritic acid) impacted growth in at least one of the algal cultures. Two of these compounds (dodecanedioic acid and aleuritic acid) produced contrasting results, increasing growth in some algae and decreasing growth in others. Together, our results reveal new groups of microalgal exometabolites, some of which could alter algal growth when provided exogenously, suggesting potential roles in allelopathy and algal interactions.
59 BASIC BIOLOGICAL SCIENCES↗
Economic and Environmental Sustainability of an Integrated Direct Air Capture System with Advanced Algal Biofuel Production
The presentation titled Economic and Environmental Sustainability of an Integrated Direct Air Capture System with Advanced Algal Biofuel Production. The background and motivation of the presentation will encompass promotion of decarbonization of the atmosphere, decouple algae production facilities from anthropogenic CO2 sources, identify key economic drivers, minimize cost and greenhouse gas emissions through process integration and optimization.
algal biofuel production↗
Mangrove peat and algae leachates elicit rapid and contrasting molecular and microbial responses in coastal waters
Abstract As sea level rises, previously sequestered blue carbon can be exported offshore as particulate or dissolved organic matter where it may be re-mineralized or sequestered. The priming effect, or interactive effects of organic matter turnover with a mixed substrate, is well described in soils, but still debated in aquatic systems. Priming may contribute to enhanced blue carbon re-mineralization in coastal environments. Here we examined mangrove-derived dissolved organic matter turnover in a lab incubation, with leachates from mangrove peat, 13 C-labeled algae, and peat+algae (primed). Particulate and dissolved organic matter were assessed; microbial metatranscriptomes were evaluated; and dissolved organic matter was characterized with high resolution mass spectrometry. Stable isotopes indicated rapid allocation of algal-derived dissolved organic matter into particulate organic matter. The algal treatment had the greatest increase in carbon dioxide, but primed and peat treatments had the greatest loss of dissolved organic carbon, greater RNA concentrations, and similar changes in total carbon dioxide. This suggests that, while total carbon dioxide did not increase under priming conditions, the addition of a peat substrate may promote microbial biomass production relative to carbon dioxide production. This work highlights that more targeted studies investigating the specific mechanisms of priming are necessary to address the molecular and microbial transformations associated with priming in aquatic systems.
54 ENVIRONMENTAL SCIENCES↗
Persistence of bacterial-mediated anti-rotifer protection in preliminary outdoor cultivation trial for Microchloropsis salina
Outdoor algal cultivation systems are susceptible to a wide variety of deleterious species. In previously published studies, we observed protection using microbial consortia at laboratory scale cultures; Microchloropsis salina in the presence of microbial consortia were protected from grazing from the marine rotifer, Brachionus plicatilis. Our objective for the present work was to determine if this protection conferred by microbial consortia in controlled laboratory experiments would persist in an open, outdoor multi-liter cultivation system. We found that algal protection did persist as evidenced by the presence of fewer motile rotifers and decreased rotifer-associated egg counts for the consortia-treated outdoor cultures. Due to the low temperature and light conditions that reduced growth of the algae outdoors, we performed an indoor laboratory assay which also confirmed the persistence of algal protection. Lastly, the lower numbers of motile rotifers and fewer rotifer-associated eggs in the consortia-treated algal cultures suggests a possible protective mechanism by the consortia through interfering with the rotifer lifecycle or reproduction. Finally, these initial results support the possibility that low cost, prophylactic treatments with microbial consortia can protect algae from deleterious species in outdoor cultivation systems.
59 BASIC BIOLOGICAL SCIENCES↗
Systems Analysis and Engineering of Biofuel Production in Chromochloris zofingiensis , an Emerging Model Green Alga
As a core component of a sustainable bioeconomy, microalgae have the potential to become a major source of biofuels and bioproducts. These photosynthetic microbes utilize solar energy, grow quickly, consume CO 2 , and can be cultivated on non-arable land. However, there are presently considerable practical limitations in the photosynthetic production of biofuels from microalgae, resulting in low productivity and high costs. Algae are a strikingly diverse group and understudied algae can reveal new opportunities for biofuels. Integrative systems biology and engineering of emerging model systems are needed to expand the possibilities of microbial production of biofuels and bioproducts. Our long-term goal is to design and engineer high-level production of biofuel precursors in microalgae.
09 BIOMASS FUELS↗
Process Optimization and Real-Time Control of Synergistic Microalgae Cultivation and Wastewater Treatment (Final Technical Report)
The overarching goal of this work was to accelerate the commercialization of high productivity, mixed community microalgal treatment technologies for the synergistic treatment of wastewater and the production of biofuel feedstocks. This project addressed a critical barrier to the financial viability and energy efficiency of algal wastewater treatment: an inability to design and operate high-rate processes that reliably achieve target effluent qualities, areal productivities, and biochemical compositions (lipid, protein, carbohydrate content) despite fluctuations in wastewater composition, weather, and microbial communities. Key outcomes from this work include an optimized and controlled Advanced Biological Nutrient Recovery (ABNR) design as well as a suite of open-source tools that include a calibrated and validated algae process simulator in QSDsan and a novel low-cost, real-time microbial monitoring tool. These tools can be leveraged by other algal cultivation and wastewater treatment technology developers in future work.
09 BIOMASS FUELS↗