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DOE OSTI2012

A comparison of Nannochloropsis salina growth performance in two outdoor pond designs: conventional raceways versus the ARID pond with superior temperature management

The present study examines how climatic conditions and pond design affect the growth performance of microalgae. From January to April of 2011, outdoor batch cultures of Nannochloropsis salina were grown in three replicate 780 L conventional raceways, as well as in an experimental 7500 L ARID (Algae Raceway Integrated Design) pond. The ARID culture system utilizes a series of 8 to 20 cm deep basins and a 1.5 m deep canal to enhance light exposure and mitigate temperature variations and extremes. The ARID culture reached the stationary phase 27 days earlier than the conventional raceways, which can be attributed to its superior temperature management and shallower basins. On a night when the air temperature dropped to -9 °C, the water temperature was 18 °C higher in the ARID pond than in the conventional raceways. Lipid and fatty acid content ranged from 16 - 25 % and 5 - 15 %, respectively, as a percentage of AFDW. Palmitic, palmitoleic, and eicosapentaenoic acid comprised the majority of fatty acids. While the ARID culture system achieved nearly double the volumetric productivity relative to the conventional raceways (0.023 vs 0.013 g L-1day-1), areal biomass productivities were of similar magnitude in both pond systems (3.34 vs. 3.47 g m-2day-1), suggesting that the ARID pond design has to be further optimized, most likely by increasing the culture depth or operating at higher cell densities while maintaining adequate mixing.

Crowe, Braden J↗

DOE OSTI2018

Valine-glutamine (VQ) motif coding genes are ancient and non-plant-specific with comprehensive expression regulation by various biotic and abiotic stresses

Valine-glutamine (VQ) motif containing proteins play important roles in abiotic and biotic stress responses in plants. However, little is known about the origin and evolution as well as comprehensive expression regulation of the VQ gene family. In this study, we systematically surveyed this gene family in 50 plant genomes from algae, moss, gymnosperm and angiosperm and explored their presence in other species from animals, bacteria, fungi and viruses. No VQs were detected in all tested algae genomes and all genomes from moss, gymnosperm and angiosperm encode varying numbers of VQs. Interestingly, some of fungi, lower animals and bacteria also encode single to a few VQs. Thus, they are not plant-specific and should be regarded as an ancient family. Their family expansion was mainly due to segmental duplication followed by tandem duplication and mobile elements. Limited contribution of gene conversion was detected to the family evolution. Generally, VQs were very much conserved in their motif coding region and were under purifying selection. However, positive selection was also observed during species divergence. Many VQs were up- or down-regulated by various abiotic / biotic stresses and phytohormones in rice and Arabidopsis. They were also co-expressed with some of other stress-related genes. All of the expression data suggest a comprehensive expression regulation of the VQ gene family. We provide new insights into gene expansion, divergence, evolution and their expression regulation of this VQ family. VQs were detectable not only in plants but also in some of fungi, lower animals and bacteria, suggesting the evolutionary conservation and the ancient origin. Overall, VQs are non-plant-specific and play roles in abiotic / biotic responses or other biological processes through comprehensive expression regulation.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Nomenclatural issues concerning cultured yeasts and other fungi: why it is important to avoid unneeded name changes

The unambiguous application of fungal names is important to communicate scientific findings. Names are critical for (clinical) diagnostics, legal compliance, and regulatory controls, such as biosafety, food security, quarantine regulations, and industrial applications. Consequently, the stability of the taxonomic system and the traceability of nomenclatural changes is crucial for a broad range of users and taxonomists. The unambiguous application of names is assured by the preservation of nomenclatural history and the physical organisms representing a name. Fungi are extremely diverse in terms of ecology, lifestyle, and methods of study. Predominantly unicellular fungi known as yeasts are usually investigated as living cultures. Methods to characterize yeasts include physiological (growth) tests and experiments to induce a sexual morph; both methods require viable cultures. Thus, the preservation and availability of viable reference cultures are important, and cultures representing reference material are cited in species descriptions. Historical surveys revealed drawbacks and inconsistencies between past practices and modern requirements as stated in the International Code of Nomenclature for Algae, Fungi, and Plants (ICNafp). Improper typification of yeasts is a common problem, resulting in a large number invalid yeast species names. With this opinion letter, we address the problem that culturable microorganisms, notably some fungi and algae, require specific provisions under the ICNafp. We use yeasts as a prominent example of fungi known from cultures. But viable type material is important not only for yeasts, but also for other cultivable Fungi that are characterized by particular morphological structures (a specific type of spores), growth properties, and secondary metabolites. We summarize potential proposals which, in our opinion, will improve the stability of fungal names, in particular by protecting those names for which the reference material can be traced back to the original isolate.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2014

Deep RNA Sequencing Reveals Hidden Features and Dynamics of Early Gene Transcription in Paramecium bursaria Chlorella Virus 1

Paramecium bursaria chlorella virus 1 (PBCV-1) is the prototype of the genus Chlorovirus (family Phycodnaviridae) that infects the unicellular, eukaryotic green alga Chlorella variabilis NC64A. The 331-kb PBCV-1 genome contains 416 major open reading frames. A mRNA-seq approach was used to analyze PBCV-1 transcriptomes at 6 progressive times during the first hour of infection. The alignment of 17 million reads to the PBCV-1 genome allowed the construction of single-base transcriptome maps. Significant transcription was detected for a subset of 50 viral genes as soon as 7 min after infection. By 20 min post infection (p.i.), transcripts were detected for most PBCV-1 genes and transcript levels continued to increase globally up to 60 min p.i., at which time 41% or the poly (A+)-containing RNAs in the infected cells mapped to the PBCV-1 genome. For some viral genes, the number of transcripts in the latter time points (20 to 60 min p.i.) was much higher than that of the most highly expressed host genes. RNA-seq data revealed putative polyadenylation signal sequences in PBCV-1 genes that were identical to the polyadenylation signal AAUAAA of green algae. Several transcripts have an RNA fragment excised. However, the frequency of excision and the resulting putative shortened protein products suggest that most of these excision events have no functional role but are probably the result of the activity of misled splicesomes.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2026

Mono-mix strategy enables comparative proteomics of a cross-kingdom microbial symbiosis

Cross-kingdom microbial symbioses, such as those between algae and bacteria, are key players in biogeochemical cycles. The molecular changes during initiation and establishment of symbiosis are of great interest, but quantitatively monitoring such changes can be challenging, particularly when the microorganisms differ greatly in size or are intimately associated. Here, we analyze output from label-free, data-dependent acquisition (DDA) LC-MS/MS proteomics experiments investigating the well-studied interaction between the alga Chlamydomonas reinhardtii and the heterotrophic bacterium Mesorhizobium japonicum. We found that detection of bacterial proteins decreased in coculture by 50% proteome-wide due to the abundance of algal proteins. As a result, standard differential expression analysis led to numerous false-positive reports of significantly downregulated proteins, where it was not possible to distinguish meaningful biological responses to symbiosis from artifacts of the reduced protein detection in coculture relative to monoculture. We show that data normalization alone does not eliminate the impact of altered detection on differential expression analysis of the cross-kingdom symbiosis. We assessed two additional strategies to overcome this methodological artifact inherent to DDA proteomics. In the first, we combined algal and bacterial monocultures at a relative abundance that mimicked the coculture, creating a “mono-mix” control to which the coculture could be compared. This approach enabled comparable detection of bacterial proteins in the coculture and the monoculture control. In the second strategy, we enhanced detection of lowly abundant bacterial proteins by using sample fractionation upstream of LC-MS/MS analysis. When these simple approaches were combined, they allowed for meaningful comparisons of nearly 10,000 algal proteins and over 4,000 bacterial proteins in response to symbiosis by DDA. They successfully recovered expected changes in the bacterial proteome in response to algal coculture, including upregulation of sugar-binding proteins and transporters. They also revealed novel proteomic responses to coculture that guide hypotheses about algal-bacterial interactions.

Dupuis, Sunnyjoy [University of California, Berkel↗

DOE OSTI2021

Developments in algal processing

Macroalgae and microalgae, including cyanobacteria, are primarily photosynthetic organisms grown for the production of renewable biomass for foods, feedstocks, and high-value specialty chemicals. While there are differences in cultivating and harvesting macro- and microalgae due to scale, many of the same downstream techniques can be used for processing algal biomass. This chapter will outline various developments in processing technologies used for the treatment of algal biomass with discussion of scalability, cost, time, and efficacy. Topics will cover methods for harvesting and dewatering algae, drying algal biomass, biomass disruption, and nutrient recovery, including case studies with lessons learned. Here, the final section presents biomass applications and product considerations. While there is no universally-adopted approach for processing algal biomass, these studies provide the foundation for making informed decisions, considering the unique properties of the algae and the integrity of the desired end products.

09 BIOMASS FUELS↗

DOE OSTI2013

NADPH: Protochlorophyllide Oxidoreductase-Structure, Catalytic Function, and Role in Prolamellar Body Formation and Morphogenesis

The biosynthesis of chlorophyll is a critical biochemical step in the development of photosynthetic vascular plants and green algae. From photosynthetic bacteria (cyanobacteria) to algae, non-vascular plants, gymnosperms and vascular plants, mechanisms have evolved for protochlorophyllide reduction a key step in chlorophyll synthesis. Protochlorophyllide reduction is carried out by both a light-dependent (POR) and light-independent (LIPOR) mechanisms. NADPH: protochlorophyllide oxidoreductase (EC 1.3.1.33, abbreviated POR) catalyzes the light-dependent reduction of protochlorophyllide (PChlide) to chlorophyllide (Chlide). In contrast, a light-independent protochlorophyllide reductase (LIPOR) involves three plastid gene products (chlL, chlN, and chlB) and several nuclear factors. Our work focused on characterization of both the POR and LIPOR catalyzed processes.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2014

Energy Technologies Research and Education Initiative

For this project, the intended goal of the microgrid component was to investigate issues in policy and technology that would drive higher penetration of renewable energy, and to demonstrate implementation in a utility system. The work accomplished on modeling the dynamics of photovoltaic (PV) penetration can be expanded for practical application. Using such a tool those involved in public policy can examine what the effect of a particular policy initiative, e.g., renewable portfolio standards (RPS) requirements, might be in terms of the desired targets. The work in the area of microgrid design, protection, and operation is fundamental to the development of microgrids. In particular the “Energy Delivery” paradigm provides new opportunities and business models for utilities. Ultimately, Energy Delivery could accrue significant benefits in terms of costs and resiliency. The experimental microgrid will support continued research and allow the demonstration of technology for better integration of renewables. The algal biofuels component of the project was developed to enhance the test facility and to investigate the technical and economic feasibility of a commercial-scale geothermal algal biofuels operation for replication elsewhere in the arid Southwest. The project was housed at New Mexico State University’s (NMSU’s) Geothermal Aquaculture Facility (GAF) and a design for the inoculation train and algae grow-out process was developed. The facility was upgraded with modifications to existing electrical, plumbing and structural components on the GAF and surrounding grounds. The research work was conducted on biomass-processing, harvesting, dewatering, and extraction. Additionally, research was conducted to determine viability of using low-cost, wastewater from municipal treatment plants in the cultivation units as make-up water and as a source of nutrients, including nitrogen and soluble phosphorus. Data was collected on inputs and outputs, growth evaluation and chemical composition of algal biomass feedstock. Also, research was completed on evaluating inoculation train, algae grow-out units, indoor cultivation units and the algal biomass dewatering units to identify opportunities for recapturing waste energy, water and biomass in the individual units or any combination of these units. A preliminary economic analysis was conducted.

09 BIOMASS FUELS↗

DOE OSTI2013

Development of Renewable Biofuels Technology by Transcriptomic Analysis and Metabolic Engineering of Diatoms

There is enormous interest in developing renewable sources of liquid fuels because of depletion of fossil fuel reserves, dependence on foreign sources, and increasing atmospheric CO 2 levels. Algae produce neutral lipids that are readily converted into liquid fuels such as biodiesel or JP-8 equivalent, and are attractive sources because they are far more productive than plants (yielding 10 -100’s of time more lipid per land area), and can be grown on non-cultivatable land with non-potable (brackish or salt) water sources. Unicellular algae known as diatoms were the most thoroughly characterized species in the National Renewable Energy Laboratory’s Aquatic Species Program, whose goal was to develop microalgae as renewable fuel sources. Lipid accumulation in microalgae is generally induced by nutrient limitation, which involves a change in environmental conditions. Intrinsic variability in cellular response to environmental changes prevents a high degree of control over the process. Nutrient limitation also inhibits biomass accumulation; therefore a tradeoff between high biomass and lipid production occurs. The goal of this project was to develop metabolic engineering approaches for diatoms to enable induction of lipid accumulation by controllable manipulation of intracellular processes rather than from external environmental conditions, and to manipulate carbon partitioning within the cell between lipid and carbohydrate synthesis to enable both abundant biomass and lipid accumulation. There were two specific objectives for this project; Objective 1:To perform comparative transcriptomic analysis in T. pseudonana and C. cryptica of lipid accumulation resulting from silicon and nitrogen limitation, to identify common and key regulatory steps involved in controlling lipid accumulation and carbon partitioning; and Objective 2: To metabolically engineer the cell to alter carbon partitioning to either trigger lipid induction without the need for nutrient limitation, or to enable lipid accumulation along with high biomass accumulation.The significance of this project is that it will enable greater control over lipid production in diatoms by manipulable intracellular processes rather than from variable environmental conditions, and it will possibly enable lipid accumulation under normal growth conditions. Current economics dictate the use of open outdoor raceway pond systems for commercial-scale microalgal growth for biofuels production (although advanced design enclosed bioreactors are under consideration, they are currently not cost effective). Outdoor systems are subject to large variability in environmental conditions. In microalgae, lipid accumulation generally occurs under nutrient limiting conditions, which prevents high biomass accumulation. Potentially, one could carefully adjust the level of a particular nutrient so that it would become limiting after sufficient biomass accumulated; however, given the variability inherent in microalgal cellular metabolism under different light, temperature, and nutrient regimes, this will be a relatively uncontrolled and poorly reproducible approach. A better strategy would be to provide ample nutrients, but trigger lipid accumulation “artificially” by manipulating intracellular processes through metabolic engineering. In addition, identifying the key regulatory steps involved in controlling carbon partitioning in the cell coupled with metabolic engineering should enable greater partitioning of carbon into lipids during non-limiting nutrient growth conditions. The approaches outlined in this proposal are aimed at achieving these goals, and are expected to have a substantial impact on the development of renewable biofuels technology. Development of the approaches described in this proposal will provide a rich interdisciplinary educational experience for high school and undergraduate students to foster their development in a scientific career.

09 BIOMASS FUELS↗

DOE OSTI2016

Consequences of Modification of Photosystem Stoichiometry and Amount in Cyanobacteria

The proposed research seeks to address two interconnected, important questions that impact photosynthetic processes and that reflect key differences between the photosynthetic systems of cyanobacteria and plants or algae. The first question is what are the reasons and consequences of the high photosystem I / photosystem II (PS I/PS II) ratio in many cyanobacteria, vs. a ratio that is close to unity in many plants and algae. The corresponding hypothesis is that most of PS I functions in cyclic electron transport, and that reduction in PS I will result primarily in a shortage of ATP rather than reducing power. This hypothesis will be tested by reducing the amount of PS I by changing the promoter region of the psaAB operon in the cyanobacterium Synechocystis sp. PCC 6803 and generating a range of mutants with different PS I content and thereby different PS I/PS II ratios, with some of the mutants having a PS II/PS I ratio closer to that in plants. The resulting mutants will be probed in terms of their growth rates, electron transfer rates, and P700 redox kinetics. A second question relates to a Mehler-type reaction catalyzed by two flavoproteins, Flv1 and Flv3, that accept electrons from PS I and that potentially function as an electron safety valve leading to no useful purpose of the photosynthesis-generated electrons. The hypothesis to be tested is that Flv1 and Flv3 use the electrons for useful purposes such as cyclic electron flow around PS I. This hypothesis will be tested by analysis of a mutant strain lacking flv3, the gene for one of the flavoproteins. This research is important for a more detailed understanding of the consequences of photosystem stoichiometry and amounts in a living system. Such an understanding is critical for not only insights in the regulatory systems of the organism but also to guide the development of biological or bio-hybrid systems for solar energy conversion into fuels.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2017

2016 Billion-Ton Report: Advancing Domestic Resources for a Thriving Bioeconomy, Volume 2: Environmental Sustainability Effects of Select Scenarios from Volume 1

With the goal of understanding environmental effects of a growing bioeconomy, the U.S. Department of Energy (DOE), national laboratories, and U.S. Forest Service research laboratories, together with academic and industry collaborators, undertook a study to estimate environmental effects of potential biomass production scenarios in the United States, with an emphasis on agricultural and forest biomass. Potential effects investigated include changes in soil organic carbon (SOC), greenhouse gas (GHG) emissions, water quality and quantity, air emissions, and biodiversity. Effects of altered land-management regimes were analyzed based on select county-level biomass-production scenarios for 2017 and 2040 taken from the 2016 Billion-Ton Report: Advancing Domestic Resources for a Thriving Bioeconomy (BT16), volume 1, which assumes that the land bases for agricultural and forestry would not change over time. The scenarios reflect constraints on biomass supply (e.g., excluded areas; implementation of management practices; and consideration of food, feed, forage, and fiber demands and exports) that intend to address sustainability concerns. Nonetheless, both beneficial and adverse environmental effects might be expected. To characterize these potential effects, this research sought to estimate where and under what modeled scenarios or conditions positive and negative environmental effects could occur nationwide. The report also includes a discussion of land-use change (LUC) (i.e., land management change) assumptions associated with the scenario transitions (but not including analysis of indirect LUC [ILUC]), analyses of climate sensitivity of feedstock productivity under a set of potential scenarios, and a qualitative environmental effects analysis of algae production under carbon dioxide (CO 2 ) co-location scenarios. Because BT16 biomass supplies are simulated independent of a defined end use, most analyses do not include benefits from displacing fossil fuels or other products, with the exception of including a few illustrative cases on potential reductions in GHG emissions and fossil energy consumption associated with using biomass supplies for fuel, power, heat, and chemicals. Most analyses in volume 2 show potential for a substantial increase in biomass production with minimal or negligible environmental effects under the biomass supply constraints assumed in BT16. Although corn ethanol has been shown to achieve GHG emissions improvements over fossil fuels, cellulosic biomass shows further improvements in certain environmental indicators covered in this report. The harvest of agricultural and forestry residues generally shows the smallest contributions to changes in certain environmental indicators investigated. The scenarios show national-level net SOC gains. When expanding the system boundary in illustrative cases that consider biomass end use, reductions in GHG emissions are estimated for scenarios in which biomass—rather than oil, coal, and natural gas—is used to produce fuel, power, heat, and chemicals. Analyses of water quality reveal that there could be tradeoffs between biomass productivity and some water quality indicators, but better outcomes for both biomass productivity and water quality can be achieved with selected conservation practices. Biodiversity analyses show possible habitat benefits to some species, with other species showing potential adverse effects that may require additional safeguards. Increasing productivity of algae can reduce GHG emissions and water consumption associated with producing algal biomass, though the effects of water consumption are likely of greater concern in some regions than in others. Moreover, the effects of climate change on potential biomass production show gains and losses in yield among feedstocks across the continental United States. Key research gaps and priorities include actions that can enhance benefits and reduce potential for negative effects of increased biomass production. The results from this report will help DOE, the bioenergy industry, and other institutions continue important discussions on environmental effects and will help chart a path toward a more environmentally sustainable bioeconomy.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2013

Feedstock to Tailpipe Initiative: Kansas Biofuels Production, Testing and Certification Laboratory

The primary task of this grant was to establish an ASTM testing facility for biodiesel and ethanol and to use this facility to develop methods to predict fuel characteristics based on feedstock composition and feedstock cultivation. In addition to characterizing fuel properties, this grant allowed for the purchase and installation of a Fourier Transform Infrared Spectroscopy (FTIR) emissions analyzer that will provide an analysis of the emissions leaving the engine in order to meet EPA regulations. This FTIR system is combined with an Alternating Current (AC) dynamometer that allows the engine to follow Environmental Protection Agency (EPA) Federal Test Procedure (FTP) cycles. A secondary task was to investigate cultivating algae utilizing wastewater and top-down ecological control and subsequent harvesting using coagulation and dissolved air flotation. Lipid extraction utilizing environmentally-friendly and cost-effective solvents, with and without cell-disruption pretreatment was also explored. Significant work on the hydrothermal liquefaction of wastewater cultivated algae was conducted.

09 BIOMASS FUELS↗

DOE OSTI2017

New approaches for metabolomics by mass spectrometry

Small molecules constitute a large part of the world around us, including fossil and some renewable energy sources. Solar energy harvested by plants and bacteria is converted into energy rich small molecules on a massive scale. Some of the worst contaminants of the environment and compounds of interest for national security also fall in the category of small molecules. The development of large scale metabolomic analysis methods lags behind the state of the art established for genomics and proteomics. This is commonly attributed to the diversity of molecular classes included in a metabolome. Unlike nucleic acids and proteins, metabolites do not have standard building blocks, and, as a result, their molecular properties exhibit a wide spectrum. This impedes the development of dedicated separation and spectroscopic methods. Mass spectrometry (MS) is a strong contender in the quest for a quantitative analytical tool with extensive metabolite coverage. Although various MS-based techniques are emerging for metabolomics, many of these approaches include extensive sample preparation that make large scale studies resource intensive and slow. New ionization methods are redefining the range of analytical problems that can be solved using MS. This project developed new approaches for the direct analysis of small molecules in unprocessed samples, as well as pushed the limits of ultratrace analysis in volume limited complex samples. The projects resulted in techniques that enabled metabolomics investigations with enhanced molecular coverage, as well as the study of cellular response to stimuli on a single cell level. Effectively individual cells became reaction vessels, where we followed the response of a complex biological system to external perturbation. We established two new analytical platforms for the direct study of metabolic changes in cells and tissues following external perturbation. For this purpose we developed a novel technique, laser ablation electrospray ionization (LAESI), for metabolite profiling of functioning cells and tissues. The technique was based on microscopic sampling of biological specimens by mid-infrared laser ablation followed by electrospray ionization of the plume and MS analysis. The two main shortcomings of this technique had been limited specificity due to the lack of a separation step, and limited molecular coverage, especially for nonpolar chemical species. To improve specificity and the coverage of the metabolome, we implemented the LAESI ion source on a mass spectrometer with ion mobility separation (IMS). In this system, the gas phase ions produced by the LAESI source were first sorted according to their collisional cross sections in a mobility cell. These separated ion packets were then subjected to MS analysis. By combining the atmospheric pressure ionization with IMS, we improved the metabolite coverage. Further enhancement of the non-polar metabolite coverage resulted from the combination of laser ablation with vacuum UV irradiation of the ablation plume. Our results indicated that this new ionization modality provided improved detection for neutral and non-polar compounds. Based on rapid progress in photonics, we had introduced another novel ion source that utilized the interaction of a laser pulse with silicon nanopost arrays (NAPA). In these nanophotonic ion sources, the structural features were commensurate with the wavelength of the laser light. The enhanced interaction resulted in high ion yields. This ultrasensitive analytical platform enabled the MS analysis of single yeast cells. We extended these NAPA studies from yeast to other microorganisms, including green algae (Chlamydomonas reinhardtii) that captured energy from sunlight on a massive scale. Combining cellular perturbations, e.g., through environmental changes, with the newly developed single cell analysis methods enabled us to follow dynamic changes induced in the cells. In effect, we were able to use individual cells as a “laboratory,” and approached the long-standing goal of establishing a “lab-in-a-cell.” Model systems for these studies included cells of cyanobacteria (Anabaena), yeast (Saccharomyces cerevisiae), green algae (C. reinhardtii) and Arabidopsis thaliana.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2016

Mixo-IBR BP1A Report

The objective of the initial budget period was to determine whether BioProcess Algae (BPA) had all of the data, information, facilities, and agreements presented in its proposal response to DE-FOA-0000739. BPA was to demonstrate heterotrophic lipid production, demonstrate the production of fuels by metathesis of algal oils, and develop data to justify the scale-up effort proposed for the Pilot Mixotrophic Algae Integrated Biorefinery.

09 BIOMASS FUELS↗

DOE OSTI2017

Supply Chain Sustainability Analysis of Renewable Hydrocarbon Fuels via Indirect Liquefaction, Fast Pyrolysis, and Hydrothermal Liquefaction: Update of the 2016 State-of-Technology Cases and Design Cases

This technical memorandum describes the supply chain sustainability analyses (SCSA) for the production of three renewable hydrocarbon transportation fuels: (1) renewable high octane gasoline (HOG) via indirect liquefaction (IDL) of woody lignocellulosic biomass; (2) renewable gasoline (RG) via fast pyrolysis of woody lignocellulosic biomass; and (3) renewable diesel via hydrothermal liquefaction of algae. This technical memorandum focuses on the 2016 State of Technology (SOT) technical, economic, and environmental performance of these three fuel production pathways, as well as the 2016 SOT woody feedstock blend production and the 2016 SOT for algae feedstock production. The results of these renewable hydrocarbon fuel pathways in these SCSA analyses update those for the respective 2015 SOT cases (Cai et al., 2016; Adom et al., 2016; Frank et al., 2016), and provide an opportunity to examine the impact of technology improvements of both biomass feedstock production and biofuel production that have been achieved since the 2015 SOTs on the sustainability performance of these renewable transportation fuels. Furthermore, they reflect updates to Argonne National Laboratory’s Greenhouse gases, Regulated Emissions, and Energy use in Transportation (GREET®) model, which was released in October 2016 (ANL, 2016). The 2015 SOT case was re-evaluated using this newly-released version, which includes updates to the production of natural gas, electricity, and liquid fuels that can influence biofuels’ supply chain greenhouse gas (GHG) (CO2, CH4, and N2O) emissions. These emissions and water consumption are the two sustainability metrics assessed in this analysis. The design cases (future target projections) for these three fuel production pathways were also re-evaluated using GREET 2016. In particular, the 2016 woody blend feedstock SOT was used in lieu of the previous design case woody blend feedstock, which utilized construction and demolition waste (INL, 2014). In the 2016 woody blend feedstock SOT, the construction and demolition waste was eliminated due to a lack of Renewable Identification Number (RINs) generation, and replaced with short rotation hybrid poplar in order to align with the woody feedstock blend that was chosen by the BETO Feedstock-Conversion Interface Consortium (FCIC) for the 2017 fast pyrolysis verification.

09 BIOMASS FUELS↗

DOE OSTI2019

Considerations for the Design of a Gas Transport System for Co-location of Microalgae Cultivation with CO 2 Sources

Co-locating microalgae cultivation facilities with sources of waste CO 2 may present opportunities for cost savings that could benefit the algal biomass industry and industries or utilities that have incentives to manage carbon emissions. However, the cost savings have not been quantified. We compare the cost of utilizing CO 2 from flue gas transported to 405-ha, base-case, open-pond microalgae facilities to the commercial purchase price of CO 2 . Sources of CO 2 include coal- and natural gas-fired power plants and ethanol, ammonia, and cement production plants in the United States. The transport of CO 2 -containing gases to the microalgae cultivation facility requires infrastructure and electricity. This engineering analysis explores the parameters that affect the infrastructure and transport distance over which emitted flue gases can be cost-effectively transported to microalgae cultivation facilities, compared to the cost of commercially purchased CO 2 (i.e., the break-even distance). Parameters that are varied include size of facility pond area, productivity, and daily duration of waste CO 2 emissions that are transported to algae. Under our assumptions and under all cases in this study, cost savings can be achieved by co-locating microalgae facilities near waste CO 2 sources. The break-even distance, which is estimated here to range from under 2 to 80 km, depends primarily on the concentration of CO 2 in flue gas, algae productivity, facility size, and pipeline system design considerations. Greater break-even distances are simulated if electricity cost is minimized, rather than if capital cost is minimized.

09 BIOMASS FUELS↗

DOE OSTI2019

Atmospheric CO 2 Capture and Membrane Delivery (Final Report)

Motivation: A major bottleneck for growing microalgae as a sustainable alternative to fossil carbon in economically producing fuels and chemical products is the cost of delivering CO 2 in sufficient concentrations for it not to limit growth. This project sought to develop and integrate two innovative technologies for capturing and concentrating CO 2 from air and delivering it to microalgae with high efficiency into an Atmospheric CO 2 Enrichment and Delivery (ACED) system. The CO 2 capture technology is based on moisture swing sorption (MSS), where specialized resin materials selectively capture CO 2 when dry and release it when wet into a confined space where the concentration can be increased up to 500-fold. The CO 2 -delivery technology is based on membrane carbonation (MC), which uses hollow fiber membranes that allow CO 2 to diffuse into the algae-containing liquid without forming bubbles, achieving nearly 100% delivery efficiency. Work completed: 1. Subsystem design, construction, and characterization [Months 1-6]. ACED integration requirements were defined; prototype MSS and MC sub-systems were constructed to conform to these requirements and characterized independently. 2. Subsystem optimization and evaluation [Months 7-12]. An iterative process of operating and upgrading each subsystem at lab scale, leading to compatibility at full integration. 3. System Integration with open raceway, testing, and modeling [Months 13-24]. Operation and testing of the integrated ACED system in an open raceway pond and modeling. Broader impacts: The ACED research program added to our understanding of 1) MC technology’s (i.e., hollow fiber membranes) utility for transferring pure CO 2 and CO 2 gas mixtures into microlagal cultures in laboratory and outdoor environments, 2) the first-of-a-kind MSS technology to capture CO 2 from ambient outdoor air, and 3) methods for storing captured CO 2 in carbonate/bicarbonate brines from which the CO 2 can be retrieved at high concentration (> 90%). Near-term commercial opportunities were outlined for the MC technology for small scale algae cultivation with high CO 2 costs (> $100/ton) and larger scale operators with continued development. This will help reduce costs for producing sustainable fuels and products and reduce cost for microalgal research for developing new applications. Research areas were identified for MSS technology to achieve cost competitive CO 2 produced from ambient air ($100/ton) as a means for closing the carbon cycle to make products from atmospheric CO 2 instead of fossil fuels and remove CO 2 from air to mitigate climate change.

09 BIOMASS FUELS↗

DOE OSTI2020

Algae/Wood Blends Hydrothermal Liquefaction and Upgrading: 2019 State of Technology

The fiscal year (FY) 2019 State of Technology (SOT) Assessment for a microalgae and woody biomass blend feedstock hydrothermal liquefaction (HTL) and biocrude upgrading system has been completed and reported here. This study is a preliminary economic analysis for this system. Inputs from the 2019 NREL open pond algae cultivation model provided microalgae seasonal flowrates and dewatered algae feedstock price. These inputs have been coupled with modeled experimental data from PNNL for algae/wood blends processed via HTL to assess the cost impacts. The TEA results demonstrated a 28% reduction in the conversion cost only from $1.22/gge in 2018 to $0.88/gge in 2019.

09 BIOMASS FUELS↗