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All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

2023 Billion-Ton Report: An Assessment of U.S. Renewable Carbon Resources - Microalgae

Microalgae is a unique biomass resource that does not need to compete for land and water with other biomass feedstocks because it can be cultivated on low-quality unencumbered land using noncompetitive water types including saline and wastewater. It is included as a complementary resource alongside other biomass feedstocks reported in this study, albeit at higher biomass production costs reflective of more capital-intensive farming operations than typical for terrestrial biomass. Higher biomass costs can be offset by the potential to produce value-added coproducts unique to compositional constituents of microalgae. Relative to the 2016 Billion-Ton Report, this chapter reflects the latest analysis from the 2022 Algae Harmonization Update, which uses the latest parameterized and high-performing saline algal strain, second-generation carbon capture of point-source waste CO 2 , and high-pressure pipeline transport resolved to specific point-source types, saline water sourcing up to 40,000 mg/L total dissolved solids for source and makeup water salinity, blowdown water treatment and recycle, and brine disposal handling. National-scale algal biomass availability potential was calculated at 152 million tons/yr ash-free dry weight (AFDW) (191 million tons/yr dry weight) at an average biomass

09 BIOMASS FUELS↗
DOE OSTI2019

Reused Cultivation Water Accumulates Dissolved Organic Carbon and Uniquely Influences Different Marine Microalgae

Reusing growth medium (water supplemented with nutrients) for microalgae cultivation is required for economical and environmentally sustainable production of algae bioproducts (fuels, feed, and food). However, reused medium often contains microbes and dissolved organic matter that may affect algae growth. While the accumulation of dissolved organic carbon (DOC) in reused medium has been demonstrated, it is unclear whether DOC concentrations affect algae growth or subsequent rates of algal DOC release. To address these questions, lab-scale experiments were conducted with three marine microalgae strains, Navicula sp. SFP, Staurosira sp. C323, and Chlorella sp. D046, grown in medium reused up to four times. Navicula sp. and Chlorella sp. grew similarly in reused medium as in fresh medium, while Staurosira sp. became completely inhibited in reused medium. Across the three algae, there was no broad trend between initial DOC concentration in reused medium and algae growth response. Navicula sp. released less DOC overall in reused medium than in fresh medium, but DOC release rates did not decrease proportionally with increased DOC concentrations. Net DOC accumulation was much lower than gross DOC released by Navicula sp. and Staurosira sp., indicating the majority of released DOC was degraded. Additionally, biodegradation experiments with reused media showed no further net decrease in DOC, suggesting the accumulated DOC was recalcitrant to the associated bacteria. Overall, these results suggest that taxa-specific factors may be responsible for algae growth response in reused medium, and that DOC release and accumulation are insensitive to prior cultivation rounds. Choosing an algae strain that is uninhibited by accumulated DOC is therefore critical to ensure successful water reuse in the algae industry.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Preservation of Microalgae, Lignocellulosic Biomass Blends by Ensiling to Enable Consistent Year-Round Feedstock Supply for Thermochemical Conversion to Biofuels

Seasonal variation in microalgae productivity is a significant barrier to economical production of algae biofuels and chemicals. Summer production can be 3–5 times higher than in the winter resulting in uneven feedstock supplies at algae biorefineries. A portion of the summer production must be preserved for conversion in the winter in order to maintain a biorefinery running at capacity. Ensiling, a preservation process that utilizes lactic acid fermentation to limit microbial degradation, has been demonstrated to successfully stabilize algae biomass (20% solids) and algae-lignocellulosic blends (40% algae-60% lignocellulosic biomass, dry basis) for over 6 months, resulting in fuel production cost savings with fewer emissions. Preservation of algae as blends could be beneficial to biorefineries that utilize thermochemical approaches to fuel production as co-processing of algae and lignocellulosic biomass has been observed to enhance biocrude yield and improve oil quality. This study conducts a resource assessment of biomass residues in the southern United States to identify materials available during peak algae productivity and in sufficient quantity to meet the algae storage needs of an algae biofuel industry. Eight feedstocks met the quantity threshold but only three, distillers grains, haylage, and yard waste, were also available in season. Storage experiments utilizing both freshwater and marine strains of microalgae – Scenedesmus acutus, Chlorella vulgaris, Chlorella zofingiensis, Nannochloropsis gaditana, and Porphyridium purpureum – and yard waste were conducted for 30 days. Storage losses were less than 10% in all but one case, and the pH of all but one blend was reduced to less than 4.7, indicating that yard waste is a suitable feedstock for blending with algae prior to storage. To better understand whether the benefits to conversion realized by processing blends might be affected by storage, elemental analysis and bomb calorimetry of pre- and post-storage algae-yard waste blends were conducted to characterize changes occurring during storage. Storing algae biomass as blends with lignocellulosic biomass could be an effective method of mitigating seasonal variability in algae biomass production while retaining the synergistic effect of co-processing algae blends in thermochemical conversion.

09 BIOMASS FUELS↗
DOE OSTI2023

Do betaine lipids replace phosphatidylcholine as fatty acid editing hubs in microalgae?

Acyl editing refers to a deacylation and reacylation cycle on a lipid, which allows for fatty acid desaturation and modification prior to being removed and incorporated into other pools. Acyl editing is an important determinant of glycerolipid synthesis and has been well-characterized in land plants, thus this review begins with an overview of acyl editing in plants. Much less is known about acyl editing in algae, including the extent to which acyl editing impacts lipid synthesis and on which lipid substrate(s) it occurs. This review compares what is known about acyl editing on its major hub phosphatidylcholine (PC) in land plants with the evidence for acyl editing of betaine lipids such as diacylglyceryltrimethylhomoserine (DGTS), the structural analog that replaces PC in several species of microalgae. In land plants, PC is also known to be a major source of fatty acids and diacylglycerol (DAG) for synthesis of the neutral lipid triacylglycerol (TAG). We review the evidence that DGTS contributes substantially to TAG accumulation in algae as a source of fatty acids, but not as a precursor to DAG. We conclude with evidence of acyl editing on other membrane lipid substrates in plants and algae apart from PC or DGTS, and discuss future analyses to elucidate the role of DGTS and other betaine lipids in acyl editing in microalgae.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Microalgae Feedstocks for Aviation Fuels

There is significant global interest in developing, testing, and using alternative jet fuels for both commercial and military use in an effort to create a sustainable and stable fuel supply while reducing greenhouse gas emissions. Currently, the aviation industry is entirely dependent on a finite-supply of petroleum based fuel sourced in part by politically and economically unstable regions of the world. Commercial jet fuel use within the contiguous United States (CONUS) was 17.8 billion gallons per year (BGY) in 2009, while jet fuel use in 2010 by the U.S. Air Force (USAF), Navy, and Army was 1.5 BGY, 0.6 BGY, and 0.8 BGY, respectively (Carter et al., 2011). U.S. commercial and military aviation sectors have set ambitious near-term alternative fuel and environmental performance targets. This includes a tentative Federal Aviation Administration (FAA) goal of 1 BGY alternative fuel use by commercial aircraft by 2018. The USAF has set a target of 50% for USAF domestic aviation via alternative fuels by 2016 (0.73 BGY), and 50% of the Navy’s total energy consumption afloat (0.3 BGY) will come from alternative fuels by 2020 (Carter et al., 2011). If these targets become policy, at least 2 BGY of domestically-produced alternative jet fuel will be required by 2020. The Energy Independence and Security Act (EISA) of 2007 established production requirements for domestic alternative fuels under the Renewable Fuel Standard (RFS). For example, 36 billion gallons of renewable fuel must be produced by 2022, of which 21 billion gallons shall be advanced biofuels. EISA defines advanced biofuels as non-corn starch derived biofuels having lifecycle greenhouse gas emissions 50% lower than gasoline. There a number of potential fuel pathways for meeting the RFS. One of these is biomass-based diesel, including jet fuel (Schnepf and Yacobucci, 2013). The U.S. Department of Energy (DOE) Bioenergy Technologies Office (BETO) has a stated goal in its 2013 Multi-Year Program Plan (USDOE, 2013) to support the RFS through development of “…commercially viable biomass utilization technologies to encourage the creation of a new domestic bioenergy industry…”. BETO has also recognized the potential for aviation biofuels to support the bioenergy industry, seeing drop-in bio-based jet fuels one of the viable alternatives for the aviation industry and the military to meet their ambitious near-term GHG reduction targets (USDOE, 2014). One of the important Multi-year Program Plan Targets (USDOE, 2013) is to establish feedstock resource assessment models to evaluate the geographic, economic, quality and environmental criteria for which 20 million metric tons of ash free dry weight (AFDW) algal biomass can be produced by 2022. Toward meeting the EISA requirements, algal biofuels may offer a number of advantages. They can produce a range of biofuel feedstocks suitable for diesel and aviation fuels. Microalgae, on a strain-specific basis, can be cultivated using impaired water including saline, and/or brackish pumped groundwater or seawater, treated industrial wastewater, municipal sewage effluent, and produced water generated from oil and gas drilling operations. Additionally, microalgae require nitrogen and phosphates as essential nutrients and could provide water treatment co-benefits to municipalities, industry, and the environment.

algae↗
DOE OSTI2022

An ensemble data assimilation modeling system for operational outdoor microalgae growth forecasting

Microalgae have received increasing attention as a potential feedstock for biofuel or biobased products. Forecasting the microalgae growth is beneficial for managers in planning pond operations and harvesting decisions. This study proposed a biomass forecasting system comprised of the Huesemann Algae Biomass Growth Model (BGM), the Modular Aquatic Simulation System in Two Dimensions (MASS2), ensemble data assimilation (DA), and numerical weather prediction Global Ensemble Forecast System (GEFS) ensemble meteorological forecasts. The novelty of this study is to seek the use of ensemble DA to improve both BGM and MASS2 model initial conditions with the assimilation of biomass and water temperature measurements and consequently improve short-term biomass forecasting skills. This study introduces the theory behind the proposed integrated biomass forecasting system, with an application undertaken in pseudo-real-time in three outdoor ponds cultured with Chlorella sorokiniana in Delhi, California, United States. Results from all three case studies demonstrate that the biomass forecasting system improved the short-term (i.e., 7-day) biomass forecasting skills by about 60% on average, comparing to forecasts without using the ensemble DA method. Given the satisfactory performances achieved in this study, it is probable that the integrated BGM-MASS2-DA forecasting system can be used operationally to inform managers in making pond operation and harvesting planning decisions.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2018

Advanced genetic tools enable synthetic biology in the oleaginous microalgae Nannochloropsis sp.

Nannochloropsis is a genus of fast-growing microalgae that are regularly used for biotechnology applications. Nannochloropsis species have a high triacylglycerol content and their polar lipids are rich in the omega-3 long-chain polyunsaturated fatty acid, eicosapentaenoic acid. Placed in the heterokont lineage, the Nannochloropsis genus has a complex evolutionary history. Genome sequences are available for several species, and a number of transcriptomic datasets have been produced, making this genus a facile model for comparative genomics. There is a growing interest in Nannochloropsis species as models for the study of microalga lipid metabolism and as a chassis for synthetic biology. Recently, techniques for gene stacking, and targeted gene disruption and repression in the Nannochloropsis genus have been developed. These tools enable gene-specific, mechanistic studies and have already allowed the engineering of improved Nannochloropsis strains with superior growth, or greater bioproduction.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Oil and gas produced water as a growth medium for microalgae cultivation: A review and feasibility analysis

Scale-up of microalgal biotechnology to provide large quantities of biofuel, lipids, and coproducts is not fully developed because of the large needs for nutrients, water, land, solar insolation, and CO 2 /carbon supplies. Wastewaters, including oil and gas produced water (PW), may supply a portion of these needs in regions with insufficient fresh water resources. PW is a challenging water resource for this use because of variable salinity, geochemical complexity, and the presence of biologically toxic components. In this paper we review PW volumes, quality, and use in media for microalgae production in the southwestern US, Australia, and Oman. We also include data from the southwestern US, referencing previously unpublished results from the National Alliance for Biofuels and Bioproducts (NAABB) consortium research project. We include a Supplementary Information section that explores cultivation of multiple microalgae species in PW and examines the carbon utilization process, all work performed in support of the NAABB field program. Strains of algae tested in the reviewed papers include Nannochloropsis , Dunalliella , Scenedesmus , and several mixed or unknown cultures. We conclude that the use of PW in algae cultivation is feasible, if the additional complexity of the water resource is accounted for in developing media formulations and in understanding metals uptake by the algae. We recommend additional work to standardize growth testing in PW, better and more thorough chemical analysis, and geochemical modeling of the PW used in media. As a result, expanded strain testing in PW media will identify improved strains tolerant of PW in algae cultivation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Evaluation of a high-moisture stabilization strategy for harvested microalgae blended with herbaceous biomass: Part I—Storage performance

Here, algal biomass is becoming increasingly attractive as a feedstock for biofuel production. However, the swing in algal biomass production between summer and winter months poses a challenge for delivering predictable, constant feedstock supply to a conversion facility. Drying is one approach for stabilizing algal biomass produced in excess during high productivity summer months for utilization during low productivity months, yet drying is energy intensive and thus costly. Wet, anaerobic storage, or ensiling, is a low-cost approach that is commonly used to preserve high moisture herbaceous feedstock. The potential for microalgae stabilization without the need for drying was investigated in this study by simulating ensiling, in which oxygen limitation drives anaerobic fermentation of soluble sugars to organic acids, dropping the pH and thereby stabilizing the material. Algal biomass, Scenedesmus obliquus , was blended with corn stover and stored in acidic, anaerobic conditions at 60% moisture (wet basis) to simulate wet storage by means of ensiling. Results demonstrate that algae and corn stover blends were successfully preserved in anaerobic, acidic conditions for 30 days with < 2% dry matter loss occurring during storage compared to 21% loss in aerobic, non-acidified conditions. Likewise, Scenedesmus obliquus stored alone at 80% moisture (wet basis) in acidified, anaerobic conditions for 30 days, resulted in dry matter losses of 6–14%, compared to 44% loss in neutral pH, anaerobic storage and 37% loss in a neutral pH, aerobically stored condition. Additional experiments were performed at a larger scale in which an algae and corn stover blend was subject to mechanical oxygen exclusion and a Lactobacillus acidophilus inoculum, resulting in 8% loss over 35 days and further indicating that acidic, anaerobic conditions can stabilize microalgae biomass. In summary, the stabilization of harvested algae can be achieved through anaerobic storage, securing a feedstock that is labile yet of high value.

09 BIOMASS FUELS↗
DOE OSTI2023

An attached microalgae platform for recycling phosphorus through biologically mediated fertilizer formation and biomass cultivation

Nutrient management is a global challenge for protecting water bodies from eutrophication and for retaining and sustainably recycling phosphorus within the biosphere. This challenge is especially important for water resource recovery facilities (WRRFs) in jurisdictions that limit nutrient loads in plant effluent. A microalgae-based biofilm platform has been designed, constructed, and tested to remove phosphorus and nitrogen from anaerobic digester (AD) effluent filtrate through cultivating biomass and inducing the precipitation of a mineral called struvite (NH₄MgPO₄ · 6H₂O) using a rotating algae biofilm reactor (RABR). RABRs function by rotating a growth substratum through nutrient rich water and then into the atmosphere, immersing microalgae in both sunlight and water. Photosynthesis is utilized in RABR operation to enhance struvite formation by increasing the pH value within the biofilm through the uptake of carbon dioxide from solution. Measurements of pH trended higher with depth through the biofilm when exposed to light confirming the function of photosynthesis in increasing pH and, as a consequence, struvite formation. In addition, reducing RPM allows more time for water to evaporate through exposure of the biofilm to the atmosphere and provides a management option to exceed struvite solubility product. Struvite precipitation was predicted based on chemical analysis and MINTEQ modeling of ADE, was confirmed using Scanning Electron Microscopy and Energy Dispersive X-ray Spectroscopy, and quantified by determining ash content. Increase in pH within the biofilm was confirmed as photosynthetic photon flux density increased. While struvite precipitation and removal from wastewater is conventionally accomplished through physicochemical methods, this research is the first report of struvite enhanced formation through algae biofilm-based photosynthesis processes.

09 BIOMASS FUELS↗
DOE OSTI2016

Copper uptake, intracellular localization, and speciation in marine microalgae measured by synchrotron radiation X-ray fluorescence and absorption microspectroscopy

Metal toxicity to aquatic organisms depends on the speciation of the metal and its binding to the critical receptor site(s) (biotic ligand) of the organism. The intracellular nature of the biotic ligand for Cu in microalgal cells was investigated using the high elemental sensitivity of microprobe synchrotron radiation X-ray fluorescence (SR-XRF) and X-ray absorption near-edge spectroscopy (XANES). The marine microalgae, Ceratoneis closterium , Phaeodactylum tricornutum , and Tetraselmis sp. were selected based on their varying sensitivities to Cu (72-h 50% population growth inhibitions of 8–47 μg Cu/L). Intracellular Cu in control cells was similar for all three species (2.5–3.2 × 10–15 g Cu/cell) and increased 4-fold in C. closterium and Tetraselmis sp. when exposed to copper, but was unchanged in P. tricornutum (72-h exposure to 19, 40, and 40 μg Cu/L, respectively). Whole cell microprobe SR-XRF identified endogenous Cu in the central compartment (cytoplasm) of control (unexposed) cells. After Cu exposure, Cu was colocated with organelles/granules dense in P, S, Ca, and Si and this was clearly evident in thin sections of Tetraselmis sp. XANES indicated coexistence of Cu(I) and Cu(II) in control and Cu-exposed cells, with the Cu ligand (e.g., phytochelatin) in P. tricornutum different from that in C. closterium and Tetraselmis sp. Here, this study supports the hypothesis that Cu(II) is reduced to Cu(I) and that polyphosphate bodies and phytochelatins play a significant role in the internalization and detoxification of Cu in marine microalgae.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2015

The effects of temperature and frequencies in ultrasound assisted extraction of phycocyanin from microalgae Spirulina sp

Microalgae Spirulina sp has been identified as source of protein and other high added value compounds. One of the compounds is phycocyanin as also known for antioxidant use. The extraction of this compound by using conventional method (soxhlet extraction) resulted low yield and longer processing time. This research was aimed to extract phycocyanin by using an extraction assisted by ultrasound irradiation. The extraction was performed by using variable of ultrasound frequency and extraction temperature and ethanol was used as a solvent. The result showed that yield of phycocyanin extracted by conventional method was 11.13% while the ultrasound irradiation could increase the yield up to 15.61% at constant frequency of 42 kHz, while the optimum temperature was obtained at 45°C. The analysis of variable interactions showed that both temperature and time has an interaction and temperature was the highest variable in increasing the yield. The conclusion of this research was the ultrasound could improve significantly the efficiency of extraction as well as activity of phycocyanin extracted from microalgae.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Ultrastructure of the Endoplasmic Reticulum in Eukaryotic Microalgae

ABSTRACT The endoplasmic reticulum (ER) is a large and highly dynamic component of the eukaryotic endomembrane system. In eukaryotic microalgae, it plays six distinct roles: (1) It envelopes the chromatin to form thenucleus. (2) It forms cisternae in the cytoplasm, some of which scaffold the synthesis of proteins destined for incorporation into membranes or for secretion. (3) It associates withGolgicisternae to scaffold the synthesis of glycosylated proteins. (4) It associates with theplasma membraneto mediate the synthesis and secretion of hydrophobic molecules. (5) It mediates the synthesis of cytoplasmiclipid bodies. (6) In lineages harboring complex plastids of red algal ancestry, it forms thechloroplast ER, which envelops the primary chloroplast envelope. In this review, these systems are illustrated using the quick‐freeze deep‐etch electron microscopy (QFDEEM) technique, which lifts up the topological configurations adopted by this gossamer system. A key finding is that in all the complex microalgae examined except dinoflagellates, the inner nuclear envelope membrane associates directly with the plastid‐contiguous membrane of the chloroplast ER at foci designated as chloroplast‐nuclear junctions. These junctions may play a role in regulating the maintenance and physiology of the complex organelles.

Microbiology↗
DOE OSTI2022

Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

Genetic compensation has been proposed to explain phenotypic differences between gene knockouts and knockdowns in several metazoan and plant model systems. With the rapid development of reverse genetic tools such as CRISPR/Cas9 and RNAi in microalgae, it is increasingly important to assess whether genetic compensation affects the phenotype of engineered algal mutants. While exploring triacylglycerol (TAG) biosynthesis pathways in the model alga Chlamydomonas reinhardtii, it was discovered that knockout of certain genes catalyzing rate-limiting steps of TAG biosynthesis, type-2 diacylglycerol acyltransferase genes (DGTTs), triggered genetic compensation under abiotic stress conditions. Genetic compensation of a DGTT1 null mutation by a related PDAT gene was observed regardless of the strain background or mutagenesis approach, for example, CRISPR/Cas 9 or insertional mutagenesis. However, no compensation was found in the PDAT knockout mutant. The effect of PDAT knockout was evaluated in a Δvtc1 mutant, in which PDAT was upregulated under stress, resulting in a 90% increase in TAG content. Knockout of PDAT in the Δvtc1 background induced a 12.8-fold upregulation of DGTT1 and a 272.3% increase in TAG content in Δvtc1/pdat1 cells, while remaining viable. These data suggest that genetic compensation contributes to the genetic robustness of microalgal TAG biosynthetic pathways, maintaining lipid and redox homeostasis in the knockout mutants under abiotic stress. This work demonstrates examples of genetic compensation in microalgae, implies the physiological relevance of genetic compensation in TAG biosynthesis under stress, and provides guidance for future genetic engineering and mutant characterization efforts.

09 BIOMASS FUELS↗
DOE OSTI2024

Utilization of microalgae for agricultural runoff remediation and sustainable biofuel production through an integrated biorefinery approach

Generally wastewater such agricultural runoff is considered a nuisance; however, it could be harnessed as a potential source of nutrients like nitrates and phosphates in integrated biorefinery context. In the current study, microalgae Chlorella sp. S5 was used for bioremediation of agricultural runoff and the leftover algal biomass was used as a potential source for production of biofuels in an integrated biorefinery context. The microalgae Chlorella sp. S5 was cultivated on Blue Green (BG 11) medium and a comprehensive optimization of different parameters including phosphates, nitrates, and pH was carried out to acquire maximum algal biomass enriched with high lipids content. Dry biomass was quantified using the solvent extraction technique, while the identification of nitrates and phosphates in agricultural runoff was carried out using commercial kits. The algal extracted lipids (oils) were employed in enzymatic trans-esterification for biodiesel production using whole-cell biomass of Bacillus subtilis Q4 MZ841642. The resultant fatty acid methyl esters (FAMEs) were analyzed using Fourier transform infrared (FTIR) spectroscopy and gas chromatography coupled with mass spectrometry (GC–MS). Subsequently, both the intact algal biomass and its lipid-depleted algal biomass were used for biogas production within a batch anaerobic digestion setup. Interestingly, Chlorella sp. S5 demonstrated a substantial reduction of 95% in nitrate and 91% in phosphate from agricultural runoff. The biodiesel derived from algal biomass exhibited a noteworthy total FAME content of 98.2%, meeting the quality standards set by American Society for Testing and Materials (ASTM) and European union (EU) standards. Furthermore, the biomethane yields obtained from whole biomass and lipid-depleted biomass were 330.34 NmL/g VS added and 364.34 NmL/g VS added , respectively. In conclusion, the findings underscore the potent utility of Chlorella sp. S5 as a multi-faceted resource, proficiently employed in a sequential cascade for treating agricultural runoff, producing biodiesel, and generating biogas within the integrated biorefinery concept.

09 BIOMASS FUELS↗
DOE OSTI2012

Manipulating Fatty Acid Biosynthesis in Microalgae for Biofuel through Protein-Protein Interactions

Microalgae are a promising feedstock for renewable fuels, and algal metabolic engineering can lead to crop improvement, thus accelerating the development of commercially viable biodiesel production from algae biomass. We demonstrate that protein-protein interactions between the fatty acid acyl carrier protein (ACP) and thioesterase (TE) govern fatty acid hydrolysis within the algal chloroplast. Using green microalga Chlamydomonas reinhardtii (Cr) as a model, a structural simulation of docking CrACP to CrTE identifies a protein-protein recognition surface between the two domains. A virtual screen reveals plant TEs with similar in silico binding to CrACP. Employing an activity-based crosslinking probe designed to selectively trap transient protein-protein interactions between the TE and ACP, we demonstrate in vitro that CrTE must functionally interact with CrACP to release fatty acids, while TEs of vascular plants show no mechanistic crosslinking to CrACP. This is recapitulated in vivo, where overproduction of the endogenous CrTE increased levels of short-chain fatty acids and engineering plant TEs into the C. reinhardtii chloroplast did not alter the fatty acid profile. These findings highlight the critical role of protein-protein interactions in manipulating fatty acid biosynthesis for algae biofuel engineering as illuminated by activity-based probes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Developments in commercial scale farming of microalgae and seaweeds

Expanding markets for microalgae and macroalgae products have led to increased development of commercial farming operations. While microalgae and macroalgae, or seaweed, have historically been harvested in many parts of globe, more recent developments seek to improve productivity, decrease production costs, increase scale, and mitigate environmental impacts of cultivation. Furthermore, this chapter highlights some of those recent developments and identifies future focus areas for research and development.

09 BIOMASS FUELS↗
DOE OSTI2019

De-Stressing Biofuel Microalgae: Leveraging Beneficial Microbiomes to Alleviate Harmful Conditions that Reduce Algal Growth

Industrial-scale production of algal biofuels is hampered by two major stressors: environmental and biological. Specifically, high light intensities and temperatures combined with microbial predators can wipe out entire crops of microalgae, which increases prices (Shurin et al., 2013). We are examining interactions between Desmodesmus sp. CO46, a species of microalgae used in biofuel production, and its microbiomes (also known as bacterial communities) that attach to its cell periphery. This work seeks to establish and monitor bacterial species that beneficially interact with Desmodesmus to enhance its growth by mitigating the negative effects of high light, high temperature, and pathogen attack. Algal cells were cultivated with samples of diluted microbiome bacteria in sterile artificial seawater (F/2 media), and algal growth was measured via chlorophyll-a fluorometry. By monitoring samples of Desmodesmus cultured with simplified microbiomes under high light cycled incubation, we find an array of algal growth responses ranging from highly beneficial to severely negative. Utilizing this library of helpful microbiomes will assist with the reliable, larger scale cultivation of Desmodesmus for biofuels and bioproducts and provide cheaper renewable energy. Future experimentation is intended to compare bulk algal dry weight between the microbiomes when grown in largescale, to see whether combining beneficial simplified microbiomes would result in an additive effect on algal growth enhancement, and to investigate more closely the specific metabolic exchanges between mutualistic bacteria and Desmodesmus. Eventually, the beneficial bacteria will also be identified through DNA sequencing.

09 BIOMASS FUELS↗