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At least 163 records · Page 9

Optimizing Selection Pressures and Pest Management to Maximize Cultivation Yield (OSPREY) (Final Technical Report)

This project was proposed in response to AOI 1, Cultivation Intensification Processes for Algae, within the FY19 Bioenergy Technologies Office Multi-Topic Funding Opportunity Announcement (FOA Number: DE-FOA-0002029). The work was designed to address a critical industry need to improve annualized productivity, stability, and quality of algal production strains for biofuels and bioproducts. The overall project goals were to generate process innovations rooted in established outdoor systems for strain selection, improvement, maintenance, and cultivation as well as pest detection and tracking. Planned advances included a 50% improvement in harvest yield based on AFDW (g m 2 d -1 ), 50% improvement in robustness based on stability metrics (e.g., high-productivity cultivation days, pond uptime), and 20% improvement in conversion yield. Individually, each of our planned process improvements (e.g., pest tracking) had the potential to increase productivity. However, to realize increases in yield at the system level, improvements to one unit’s process must be balanced against potential effects on other processes. For example, changes to strains, cultivation, and pest management developed in isolation may hurt other unit operations. Therefore, a critical success factor of the project was the integration of the pipeline components, achieved through iterative field-to- (short term) lab testing. In addition, through sustainability models, we evaluated how improvements would alter industry scenarios.

09 BIOMASS FUELS↗

Microalgae Biomass Production for Utilization of CO2 and Mitigation of Greenhouse Gas Emissions

Microalgal cultivation processes for production of foods, feeds, fuels, fertilizers and other bioproducts and wastewater treatment are being considered for reducing CO2 and other greenhouse gas emissions. Major differences between microalgae and higher plant biomass cultivation are include their potential for much higher productivities, their higher content of major (N-P-K) and minor nutrients, and the challenge of harvesting such microscopic plants. Most critical, microalgal mass cultures, unlike higher plants, currently require fertilization with concentrated sources of CO2. As practically all of the C fixed into algal biomass will be re-emitted into the atmosphere in short order, algae processes do not sequester carbon. Their potential for CO2 emissions reductions must thus be based on comparisons with current technologies for the production of competing products, such as biofuels or animal feeds, or in wastewater treatment. Further, due to economic limitations to flue gas transport (3 to 12 % CO2), as well as highly variable diurnal and seasonal CO2 utilization and limited land and water availability near most CO2 sources, only a small fraction of waste CO2 emissions will be directly utilizeable for microalgae biomass production. However, for long-distance transport, flue gas CO2 capture by chemical processes would greatly increase the costs of flue gas utilization. More concentrated, sources of CO2 from refineries, fertilizer, chemical plants, and fermentations are available, but are also very limited relative to the quantities required for commodity production. Microalgae can also be cultivated on organic wastes, which provide both nutrients (N, P, K, etc.) and organic and inorganic carbon for algal growth, reducing greenhouse gas emissions compared to conventional treatment processes. To maximize the potential for algae biomass production and CO2 utilization, direct capture of CO2 from air will be required. This could be accomplished with the algal cultivation process itself in large open-raceway ponds, through chemical and biological enhancements of CO2 transfer into algal cutures. The relative productivities, economics, greenhouse gas balances, and resource potentials of these alternative CO2 sources for large-scale production of microalgae will be reviewed.

09 BIOMASS FUELS↗

Chemical Processing of Non-Crop Plants for Jet Fuel Blends Production

The use of Biofuels has been gaining in popularity over the past few years due to their ability to reduce the dependence on fossil fuels. Biofuels as a renewable energy source can be a viable option for sustaining long-term energy needs if they are managed efficiently. We describe our initial efforts to exploit algae, halophytes and other non-crop plants to produce synthetics for fuel blends that can potentially be used as fuels for aviation and non-aerospace applications. Our efforts have been dedicated to crafting efficient extraction and refining processes in order to extract constituents from the plant materials with the ultimate goal of determining the feasibility of producing biomass-based jet fuel from the refined extract. Two extraction methods have been developed based on communition processes, and liquid-solid extraction techniques. Refining procedures such as chlorophyll removal and transesterification of triglycerides have been performed. Gas chromatography in tandem with mass spectroscopy is currently being utilized in order to qualitatively determine the individual components of the refined extract. We also briefly discuss and compare alternative methods to extract fuel-blending agents from alternative biofuels sources.

Kulis, M. J.↗

Cultivating clarity: understanding the impact of land cost assumptions on biofuel viability

The transition to sustainable energy has increased interest in biofuel production to reduce greenhouse gas emissions, decrease reliance on imported oil, and ensure energy resilience. Here, this study examines the often-overlooked impact of land cost assumptions on the economic viability of biofuel production. Using a discounted cash flow techno-economic framework, we evaluated three land cost scenarios—no land costs, land rental costs, and land purchase costs—across six bioenergy feedstocks (corn, soybeans, switchgrass, miscanthus, poplar, and microalgae) and three biofuel products (corn ethanol, soybean biodiesel, and sustainable aviation fuel) at the county level for the contiguous United States. The analysis reveals substantial variation in minimum fuel selling prices due to these scenarios. High-yield crops like algae showed low sensitivity to land costs, while low-yield crops such as soybeans were highly sensitive. Geographical differences were significant, with minimum fuel selling price increases most pronounced in high-value land regions like the Corn Belt. Case studies further illustrate the influence of local productivity and land costs on economic outcomes across the United States. These findings emphasize the importance of maintaining consistent land cost assumptions in biofuel economic assessments. By quantifying the interplay between land value, crop productivity, and economic feasibility, this study provides essential insights for policymakers and stakeholders to advance sustainable energy solutions.

09 BIOMASS FUELS↗

Improving biomass and lipid yields of Desmodesmus armatus and Chlorella vulgaris through mutagenesis and high-throughput screening

Advancement of microalgae for use as biofuel feedstock depends on bioprospecting novel strains and then applying classical strain improvement techniques to enhance desirable traits. Here we successfully demonstrate this approach using two novel strains of green algae, identified as Desmodesmus armatus and Chlorella vulgaris, to improve biomass and lipid yields. UV irradiation was utilized to generate random mutations and the resulting populations were analyzed via flow cytometry to quantify lipid fluorescence while enriching for high lipid variants using fluorescence activated cell sorting. Secondary screening was performed via in situ fluorescence staining to identify the single highest lipid producing mutants from each strain. This approach produced a Desmodesmus armatus mutant that exhibited both increased biomass productivity and higher lipid content, which doubled lipid yield from the culture. The resulting Chlorella vulgaris mutant exhibited an increase in growth only, resulting in a 33% increase in lipid yield.

09 BIOMASS FUELS↗

Engineering the Novel Extremophile Alga Chlamydomonas pacifica for High Lipid and High Starch Production as a Path to Developing Commercially Relevant Strains

Microalgae offer a compelling platform for the production of commodity products, due to their superior photosynthetic efficiency, adaptability to nonarable lands and nonpotable water, and their capacity to produce a versatile array of bioproducts, including biofuels and biomaterials. However, the scalability of microalgae as a bioresource has been hindered by challenges such as costly biomass production related to vulnerability to pond crashes during large-scale cultivation. This study presents a pipeline for the genetic engineering and pilot-scale production of biodiesel and thermoplastic polyurethane precursors in the extremophile species Chlamydomonas pacifica. This extremophile microalga exhibits exceptional resilience to high pH (>11.5), high salinity (up to 2% NaCl), and elevated temperatures (up to 42 °C). Initially, we evolved this strain to also have a high tolerance to high light intensity (>2000 μE/m 2 /s) through mutagenesis, breeding, and selection. We subsequently genetically engineered C. pacifica to significantly enhance lipid production by 28% and starch accumulation by 27%, all without affecting its growth rate. We demonstrated the scalability of these engineered strains by cultivating them in pilot-scale raceway ponds and converting the resulting biomass into biodiesel and thermoplastic polyurethanes. This study showcases the complete cycle of transforming a newly discovered species into a commercially relevant commodity production strain. This research underscores the potential of extremophile algae, including C. pacifica, as a key species for the burgeoning sustainable bioeconomy, offering a viable path forward in mitigating environmental challenges and supporting global bioproduct demands.

59 BASIC BIOLOGICAL SCIENCES↗

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↗

Gene Expression in Dunaliella tertiolecta +/- Iron

Dunaliella tertiolecta is an extremophilic, green alga from the Chlorophyte lineage. It is found in coastal marine environments around the world. D. tertiolecta can tolerate extremes of heat, light, pH, and salinity. D. tertiolecta is under development for the production biofuels and other bioproducts because it can produce large quantities of neutral lipids, and it can be grown in open raceway ponds using only the inputs of seawater and sunlight. This isolate of D. tertiolecta (UTEX LB 999) was found in Oslofjord, Norway in 1938. This accession includes an RNA-Seq analysis of D. tertiolecta cultures grown in iron-replete (1.5 µM) or iron-deficient (0 µM) media.

54 ENVIRONMENTAL SCIENCES↗

NASA's GreenLab Research Facility: A Guide for a Self-Sustainable Renewable Energy Ecosystem

There is a large gap between the production and demand for energy from alternative fuel and alternative renewable energy sources. The sustainability of humanity, as we know it, directly depends on the ability to secure affordable fuel, food, and freshwater. NASA Glenn Research Center (Glenn) has initiated a laboratory pilot study on using biofuels as viable alternative fuel resources for the field of aviation, as well as utilizing wind and solar technology as alternative renewable energy resources. The GreenLab Research Facility focuses on optimizing biomass feedstock using algae and halophytes as the next generation of renewable aviation fuels. The unique approach in this facility helps achieve optimal biomass feedstock through climatic adaptation of balanced ecosystems that do not use freshwater, compete with food crops, or use arable land. In addition, the GreenLab Research Facility is powered, in part, by alternative and renewable energy sources, reducing the major environmental impact of present electricity sources. The ultimate goal is to have a 100 percent clean energy laboratory that, when combined with biomass feedstock research, has the framework in place for a self-sustainable renewable energy ecosystem that can be duplicated anywhere in the world and can potentially be used to mitigate the shortage of food, fuel, and water. This paper describes the GreenLab Research Facility at Glenn and its power and energy sources, and provides recommendations for worldwide expansion and adoption of the facility s concept.

Bomani, B. M. McDowell↗

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

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

biofuels↗

Annotated Genome Sequence of the High-Biomass-Producing Yellow-Green Alga Tribonema minus

Here, we report the annotated genome sequence for a heterokont alga from the class Xanthophyceae. This high-biomass-producing strain, Tribonema minus UTEX B 3156, was isolated from a wastewater treatment plant in California. It is stable in outdoor raceway ponds and is a promising industrial feedstock for biofuels and bioproducts.

59 BASIC BIOLOGICAL SCIENCES↗

A bloom of a single bacterium shapes the microbiome during outdoor diatom cultivation collapse

Algae-dominated ecosystems are fundamentally influenced by their microbiome. We lack information on the identity and function of bacteria that specialize in consuming algal-derived dissolved organic matter in high algal density ecosystems such as outdoor algal ponds used for biofuel production. Here, we describe the metagenomic and metaproteomic signatures of a single bacterial strain that bloomed during a population-wide crash of the diatom, Phaeodactylum tricornutum, grown in outdoor ponds. 16S rRNA gene data indicated that a single Kordia sp. strain (family Flavobacteriaceae) contributed up to 93% of the bacterial community during P. tricornutum demise. Kordia sp. expressed proteins linked to microbial antagonism and biopolymer breakdown, which likely contributed to its dominance over other microbial taxa during diatom demise. Analysis of accompanying downstream microbiota (primarily of the Rhodobacteraceae family) provided evidence that cross-feeding may be a pathway supporting microbial diversity during diatom demise. In situ and laboratory data with a different strain suggested that Kordia was a primary degrader of biopolymers during algal demise, and co-occurring Rhodobacteraceae exploited degradation molecules for carbon. An analysis of 30 Rhodobacteraceae metagenome assembled genomes suggested that algal pond Rhodobacteraceae commonly harbored pathways to use diverse carbon and energy sources, including carbon monoxide, which may have contributed to the prevalence of this taxonomic group within the ponds. These observations further constrain the roles of functionally distinct heterotrophic bacteria in algal microbiomes, demonstrating how a single dominant bacterium, specialized in processing senescing or dead algal biomass, shapes the microbial community of outdoor algal biofuel ponds.

Kordia↗

Muconic Acid Production from P. putida Using High Protein Algae Hydrolysate

The composition of algal biomass is highly dynamic, with protein, lipid, and carbohydrate contents varying in response to nutrient and environmental conditions during cultivation. Because shifts in biomass composition are often associated with reduced biomass productivity, production costs can often increase if targeting higher biomass compositional quality (enriched in carbohydrates or lipids at reduced protein content) as input for the algal biorefinery. The optimal algal biorefinery configuration is thus a function of many factors. One of the key strengths of the Combined Algal Processing (CAP) process is the versatility of feedstocks and products produced. The concept has been demonstrated with ethanol and a variety of carboxylic acids (succinic, butyric, muconic) as coproducts along with lipid upgrading to biofuel. Modification of the approaches, processes and downstream upgrading to fuels has allowed the CAP process to reduce costs and improve efficiency. Muconic acid is a high-value, potential fermentation coproduct of interest because it can be easily converted to adipic acid, a high-volume monomer for the production of nylon and other valuable consumer plastics. As such, the production of muconic acid through CAP was explored to expand the suite of products from algal biomass and to begin exploring the valorization of high protein content biomass from rapidly grown algae biomass. We have established initial performance parameters and shown that the range of substrates consumed by the muconic acid-producing microbe, Pseudomonas putida, includes at least glucose, mannose, glycerol, and lactic acid. We achieved complete utilization of these four major hydrolysate substrates achieving productivities of 0.037 (g/L/h) from Scenedesmus obliquus and 0.029 (g/L/h) from Monoraphidium minutum hydrolysates. Final titer and process yield (mol of muconic acid per mol of substrate (molP/molS)) were 0.99 g/L and 0.42 molP/molS from S. obliquus hydrolysate and 0.75 g/L and 0.23 molP/molS from M. minutum hydrolysate, respectively.

BIOMASS FUELS↗

Alternate-Fueled Flight: Halophytes, Algae, Bio-, and Synthetic Fuels

Synthetic and biomass fueling are now considered to be near-term aviation alternate fueling. The major impediment is a secure sustainable supply of these fuels at reasonable cost. However, biomass fueling raises major concerns related to uses of common food crops and grasses (some also called "weeds") for processing into aviation fuels. These issues are addressed, and then halophytes and algae are shown to be better suited as sources of aerospace fuels and transportation fueling in general. Some of the history related to alternate fuels use is provided as a guideline for current and planned alternate fuels testing (ground and flight) with emphasis on biofuel blends. It is also noted that lessons learned from terrestrial fueling are applicable to space missions. These materials represent an update and additions to the Workshop on Alternate Fueling Sustainable Supply and Halophyte Summit at Twinsburg, OH, Oct. 17 to 18, 2007 (ref. 1).

Hendricks, R. C.↗

Alternate-Fueled Flight: Halophytes, Algae, Bio-, and Synthetic Fuels

Synthetic and biomass fueling are now considered to be near-term aviation alternate fueling. The major impediment is a secure sustainable supply of these fuels at reasonable cost. However, biomass fueling raises major concerns related to uses of common food crops and grasses (some also called "weeds") for processing into aviation fuels. These issues are addressed, and then halophytes and algae are shown to be better suited as sources of aerospace fuels and transportation fueling in general. Some of the history related to alternate fuels use is provided as a guideline for current and planned alternate fuels testing (ground and flight) with emphasis on biofuel blends. It is also noted that lessons learned from terrestrial fueling are applicable to space missions. These materials represent an update (to 2009) and additions to the Workshop on Alternate Fueling Sustainable Supply and Halophyte Summit at Twinsburg, Ohio, October 17 to 18, 2007.

Hendricks, R. C.↗

Chapter 5: Methods for Algal Protein Isolation and Proteome Analysis

Microalgae present promising feedstocks to produce renewable fuel and chemical intermediates, in part due to high storage carbon flux capacity to triacylglycerides or storage carbohydrates upon nutrient deprivation. However, the mechanism(s) governing deprivation-mediated carbon partitioning remain to be fully elucidated, limiting targeted strain engineering strategies in algal biocatalysts. Though genomic and transcriptomic analyses offer key insights into these mechanisms, active post-transcriptional regulatory mechanisms, ubiquitous in many microalgae, necessitate proteomic and post-translational (e.g., phospho- and nitroso-proteomic) analyses to more completely evaluate algal responsiveness to nutrient deprivation. Herein, we describe methods for isolating total algal protein and conducting proteomic, phosphoproteomic, and nitrosoproteomic analyses. We focus on methods deployed for the chlorophyte, Chlorella vulgaris, a model oleaginous alga with high flux to renewable fuel and chemical precursors.

BASIC BIOLOGICAL SCIENCES,BIOMASS FUELS↗

Multi-objective optimization of sustainable aviation fuel production pathways in the U.S. Corn Belt

As a potential source of low-carbon transportation energy, biofuels offer certain advantages over vehicle electrification (e.g., lower societal vulnerability to grid failures, and improved range of sustainable aviation), but also several challenges, including cost, carbon intensity, and land usage. There are also well-founded concerns that biofuel supply chains could be disrupted if extreme weather events impact feedstock yields. In this paper, we explore the use of multi-objective optimization to identify biofuel production pathways that balance cost, greenhouse gas emissions, and supply vulnerability to extreme weather. We compare the use of three different many-objective evolutionary algorithms and linear programming in optimizing biomass cultivation decisions in the U.S. Corn Belt under weather uncertainty using historical, modeled, and synthetic yield data. We consider four feedstock choices (corn, soy, switchgrass, and algae) with two land types (agricultural and marginal lands) and evaluate decisions using three alternative spatial resolutions (ranging from the USDA agricultural district level to the state level). Results show that feedstock choice is the primary driver of objective performance (i.e., the position and shape of 3D, approximate Pareto frontiers). Spatial diversification is a less effective tool in reducing exposure to weather-caused drops in crop yield.

09 BIOMASS FUELS↗