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Protein Network Signatures Associated with Exogenous Biofuels Treatments in Cyanobacterium Synechocystis sp. PCC 6803

Although recognized as a promising microbial cell factory for producing biofuels, current productivity in cyanobacterial systems is low. To make the processes economically feasible, one of the hurdles, which need to be overcome is the low tolerance of hosts to toxic biofuels. Meanwhile, little information is available regarding the cellular responses to biofuels stress in cyanobacteria, which makes it challenging for tolerance engineering. Using large proteomic datasets of Synechocystis under various biofuels stress and environmental perturbation, a protein co-expression network was first constructed and then combined with the experimentally determined protein–protein interaction network. Proteins with statistically higher topological overlap in the integrated network were identified as common responsive proteins to both biofuels stress and environmental perturbations. In addition, a weighted gene co-expression network analysis was performed to distinguish unique responses to biofuels from those to environmental perturbations and to uncover metabolic modules and proteins uniquely associated with biofuels stress. The results showed that biofuel-specific proteins and modules were enriched in several functional categories, including photosynthesis, carbon fixation, and amino acid metabolism, which may represent potential key signatures for biofuels stress responses in Synechocystis. Network-based analysis allowed determination of the responses specifically related to biofuels stress, and the results constituted an important knowledge foundation for tolerance engineering against biofuels in Synechocystis.

60 APPLIED LIFE SCIENCES↗

Biofuels and Food Security. A report by the High Level Panel of Experts on Food Security and Nutrition

In October 2011, the UN Committee on World Food Security (CFS) recommended a ''review of biofuels policies -- where applicable and if necessary -- according to balanced science-based assessments of the opportunities and challenges that they may represent for food security so that biofuels can be produced where it is socially, economically and environmentally feasible to do so''. In line with this, the CFS requested the HLPE (High Level Panel of Experts) to ''conduct a science-based comparative literature analysis taking into consideration the work produced by the FAO and Global Bioenergy Partnership (GBEP) of the positive and negative effects of biofuels on food security''. Recommendations from the report include the following. Food security policies and biofuel policies cannot be separated because they mutually interact. Food security and the right to food should be priority concerns in the design of any biofuel policy. Governments should adopt the principle: biofuels shall not compromise food security and therefore should be managed so that food access or the resources necessary for the production of food, principally land, biodiversity, water and labour are not put at risk. The CFS should undertake action to ensure that this principle is operable in the very varied contexts in which all countries find themselves. Given the trend to the emergence of a global biofuels market, and a context moving from policy-driven to market-driven biofuels, there is an urgent need for close and pro-active coordination of food security, biofuel/bioenergy policies and energy policies, at national and international levels, as well as rapid response mechanisms in case of crisis. There is also an urgent need to create an enabling, responsible climate for food and non-food investments compatible with food security. The HLPE recommends that governments adopt a coordinated food security and energy security strategy, which would require articulation around the following five axes/dimensions: Adapt to the change to global, market-driven dynamics; Address the land, water and resource implications of biofuel policies; Foster the transition from biofuels to comprehensive food-energy policies; Promote research and development; and, Develop methods and guidelines for coordinated food, Biofuels, and bio-energy policies at national and international levels.

09 BIOMASS FUELS↗

Biofuels Policies That Have Encouraged Their Production and Use: An International Perspective

This study evaluated the effectiveness of technology-push and market-pull policies in encouraging the production and use of biofuels in 15 countries including major biofuels producers. Biofuels production and consumption data for the 12-year period of 2006-2017 showed that in most of the surveyed countries, biofuels policies played an important role in developing and growing regional and national biofuels markets. Blending mandates continue to be the main market-pull policy used to create and expand biofuels markets. Countries with a mixture of market-pull and technology-push policy instruments have been most successful at increasing biofuels production and use and also developing and deploying less mature advanced biofuels. Most policies have primarily promoted the production and use of biofuels for road transport with key long-distance transport sectors (aviation and shipping) drawing less policy attention despite being significant fuel consumers and carbon emitters. While many older policies were originally developed to promote energy security, more recent policies, such as low carbon fuel standard, have reduction in the carbon intensity of transportation fuels as a primary goal. While policies have been essential in promoting the on-going growth of biofuels, they have not been sufficient to drive the level of development needed to decarbonize the transport sector.

biofuels for transport↗

Adoption of Biofuels for Marine Shipping

Biomass-derived fuels can undoubtedly play an essential role in a future of marine fuel that is more renewable and offers potential synergistic benefits when blended with petroleum fuels. Biomass-derived marine fuel would reduce sulfur content, as well as improve overall emission profiles and ultimately environmental sustainability (including decarbonization) for marine transport. For example, biofuel derived from biomass such as forest residues is expected to deliver possibly up to 90% of life cycle (or well-to-propeller) greenhouse gas reduction versus fossil equivalents, and virtually eliminate SOx emissions - potentially without any requirement for engine modifications. For the benefits of biofuels to be realized and to be impactful, a significant fraction of marine fossil fuel will need to be displaced with biofuel. The direct replacement of marine fossil fuel with a biofuel necessitates that the production volume of biofuel will be high enough to meet the demand. Furthermore, the potential of biofuel adoption also depends on the future biofuel price. Hence, the long-term price and availability of marine biofuel are two critical factors for establishing the feasibility of marine biofuel adoption for operating vessels. This study will shed light on the prospect of biofuel to be adopted for the maritime shipping sector.

air emissions↗

Agriculture, Land Use, Energy and Carbon Emission Impacts of Global Biofuel Mandates to Mid-Century

Three potential future scenarios of expanded global biofuel production are presented here utilizing the GCAM integrated assessment model. These scenarios span a range that encompasses on the low end a continuation of existing biofuel production policies to two scenarios that would require an expansion of current targets as well as an extension of biofuels targets to other regions of the world. Conventional oil use is reduced by 4-8% in the expanded biofuel scenarios, which results in a decrease of in CO2 emissions on the order of 1-2 GtCO2/year by mid-century from the global transportation sector. The regional distribution of crop production is relatively unaffected, but the biofuels targets do result in a marked increase in the production of conventional crops used for energy. Producer prices of sugar and corn reach levels about 12% and 7% above year 2005 levels, while the increased competition for land causes the price of food crops such as wheat, although not used for bioenergy in this study, to increase by 1 to 2%. The amount of land devoted to growing all food crops and dedicated bioenergy crops is increased by about 10% by 2050 in the High biofuel case, with concurrent decreases in other uses of land such as forest and pasture. In both of the expanded biofuels cases studied, there is an increase in net cumulative carbon emissions for the first couple of decades due to these induced land use changes. However, the difference in net cumulative emissions from the biofuels expansion decline by about 2035 as the reductions in energy system emissions exceed further increases in emissions from land use change. Even in the absence of a policy that would limit emissions from land use change, the differences in net cumulative emissions from the biofuels scenarios reach zero by 2050, and are decreasing further over time in both cases.

Biofuels↗

The greenhouse gas intensity and potential biofuel production capacity of maize stover harvest in the US Midwest

Agricultural residues are important sources of feedstock for a cellulosic biofuels industry that is being developed to reduce greenhouse gas emissions and improve energy independence. While the US Midwest has been recognized as key to providing maize stover for meeting near-term cellulosic biofuel production goals, there is uncertainty that such feedstocks can produce biofuels that meet federal cellulosic standards. Here, we conducted extensive site-level calibration of the Environmental Policy Integrated Climate (EPIC) terrestrial ecosystems model and applied the model at high spatial resolution across the US Midwest to improve estimates of the maximum production potential and greenhouse gas emissions expected from continuous maize residue-derived biofuels. A comparison of methodologies for calculating the soil carbon impacts of residue harvesting demonstrates the large impact of study duration, depth of soil considered, and inclusion of litter carbon in soil carbon change calculations on the estimated greenhouse gas intensity of maize stover-derived biofuels. Using the most representative methodology for assessing long-term residue harvesting impacts, we estimate that only 5.3 billion liters per year (bly) of ethanol, or 8.7% of the near-term US cellulosic biofuel demand, could be met under common no-till farming practices. However, appreciably more feedstock becomes available at modestly higher emissions levels, with potential for 89.0 bly of ethanol production meeting US advanced biofuel standards. Adjustments to management practices, such as adding cover crops to no-till management, will be required to produce sufficient quantities of residue meeting the greenhouse gas emission reduction standard for cellulosic biofuels. Considering the rapid increase in residue availability with modest relaxations in GHG reduction level, it is expected that management practices with modest benefits to soil carbon would allow considerable expansion of potential cellulosic biofuel production.

09 BIOMASS FUELS↗

Water impacts of U.S. biofuels: Insights from an assessment combining economic and biophysical models

Biofuels policies induce land use changes (LUC), including cropland expansion and crop switching, and this in turn alters water and soil management practices. Policies differ in the extent and type of land use changes they induce and therefore in their impact on water resources. We quantify and compare the spatially varying water impacts of biofuel crops stemming from LUC induced by two different biofuels policies by coupling a biophysical model with an economic model to simulate the economically viable mix of crops, land uses, and crop management choices under alternative policy scenarios. We assess the outputs of an economic model with a high-resolution crop-water model for major agricultural crops and potential cellulosic feedstocks in the US to analyze the impacts of three alternative policy scenarios on water balances: a counterfactual ‘no-biofuels policy’ (BAU) scenario, a volumetric mandate (Mandate) scenario, and a clean fuel-intensity standard (CFS) scenario incentivizing fuels based on their carbon intensities. While both biofuel policies incentivize more biofuels than in the counterfactual, they differ in the mix of corn ethanol and advanced biofuels from miscanthus and switchgrass (more corn ethanol in Mandate and more cellulosic biofuels in CFS). The two policies differ in their impact on irrigated acreage, irrigation demand, groundwater use and runoff. Net irrigation requirements increase 0.7% in Mandate and decrease 3.8% in CFS, but in both scenarios increases are concentrated in regions of Kansas and Nebraska that rely upon the Ogallala aquifer for irrigation water. Our study illustrates the importance of accounting for the overall LUC and shifts in agricultural production and management practices in response to policies when assessing the water impacts of biofuels.

09 BIOMASS FUELS↗

Utilizing data-based modeling with low life cycle GHG emissions algae biofuels for engine optimization

Aquatic microalgae are a highly promising feedstock for the production of biocrude and tailored biofuels, with distinct advantages over traditional terrestrial crops, such as reduced land use and avoidance of food production competition. However, unlocking their full potential requires the development of biofuels with low life cycle greenhouse emissions biofuels, such as algae biofuels, which can significantly reduce the environmental impact of the transportation systems without requiring a complete overhaul of existing engine technology. In this study, we employ cutting-edge data-based AI modeling techniques to optimize the performance of heavy-duty engines, with a focus on transitioning towards biofuels with low life cycle greenhouse emissions biofuels. Our methodology offers significant advantages over traditional sweep testing, enabling efficient and accurate optimization of engine performance with minimal time and resources consumption. Our findings demonstrate the potential of utilizing this approach, with up to 55% NOx emissions reductions and up to 2% reduction in fuel consumption compared to the baseline optimized point. Moving forward, we plan to utilize a 30% blend of algae biofuels with diesel fuel, with the ultimate goal of achieving up to 60% lifecycle GHG emissions. Lastly, we plan to compare the results with 100% renewable biodiesel to add an additional dimension of investigating the impact of fuel chemistry on engine optimization. Overall, this study underscores the vital importance of biofuels for reducing the carbon footprint of the transportation sector and supporting a sustainable future. By harnessing the power of data-based AI modeling with low life cycle greenhouse emissions biofuels, we can accelerate the adoption of more environmentally friendly transportation systems and reduce their impact on the planet. Our findings contribute to this transition and offer insights for developing efficient and effective strategies for addressing global climate change.

09 BIOMASS FUELS↗

Improved Biofuel Production through Discovery and Engineering of Terpene Metabolism in Switchgrass

Project Objectives - Of the myriad specialized metabolites that plants deploy to adapt to environmental challenges, terpenes form the largest group. In many major crops, unique terpene blends serve as key stress defenses that directly impact plant fitness and yield. In addition, terpenes, such as bisabolene and pinene, are used for producing renewable biofuels. Essential to advancing a broader use of terpenes for biofuel feedstock engineering is a system-wide knowledge of the diverse biosynthetic machinery and defensive potential of often species-specific terpene blends. The proposed project would merge genome-wide enzyme discovery with comparative –omics, protein structural and plant microbiome studies to define the biosynthesis and stress-defensive functions of the switchgrass (Panicum virgatum) terpene network. These insights would be combined with developing and applying non-transgenic genome editing tools to design plants with desirable terpene blends for higher productivity and biofuel production on marginal lands. As a dedicated lignocellulosic feedstock for U.S. biofuel production with high net energy yield, stress tolerance, and available genome resources, switchgrass is well-suited for devising new avenues for biofuel production. Project Description – The diversity of plant terpene defenses is governed by species-specific families of terpene synthase (TPS) and cytochrome P450 monooxygenase (P450) enzymes. Mining of the switchgrass genome (genotype Alamo) identified ~100 TPS and P450 candidate genes, and combinatorial biochemical analysis of synthesized TPSs and P450s revealed more than a dozen enzymes with common and novel activities. In addition, several identified terpene metabolites and the corresponding transcripts were up-regulated in response to abiotic stressors. These findings demonstrate a unique switchgrass terpene network with probable importance to abiotic stress tolerance, thus providing a large chemical portfolio for optimizing crop resistance, yield, and biofuel composition. Leveraging these preliminary data, we propose to generate a genome-wide map of the switchgrass terpene metabolic network through multi-gene co-expression analyses that allow the efficient cross-validation of TPS and P450 functions. Key enzymes would further be applied to structure-function studies via X-ray protein crystallography, homology modeling and site-directed mutagenesis to gain mechanistic insight into the catalytic specificity of switchgrass terpene metabolism and provide gene and amino acid targets for genome editing. In tandem with terpene pathway discovery, system-wide metabolomics, transcriptomics and proteomics studies in switchgrass accessions of contrasting drought tolerance would define the role of switchgrass terpene metabolism in conferring abiotic stress resilience. Metabolic changes would be assessed in a combined approach of targeted (terpenes) and untargeted metabolite profiling using a high-resolution LC-MS/MS approach, differential gene expression analyses through multiplexed Illumina RNA sequencing, and quantitative analysis of high-priority pathway enzymes using multiple reaction monitoring (MRM). Drawing on these insights, knock-down/out mutants of stress-associated pathway nodes would be generated by optimizing transient virus-induced gene silencing (VIGS) and CRISPR/Cas9 systems under control of the Tobacco Rattle Virus (TRV). The resulting mutant lines would then be analyzed for stress susceptibility and the impact on the root microbiome to define gene functions in planta. Knowledge of terpene pathways, enzyme mechanisms and bioactivities would be applied to enhance switchgrass stress resilience and to tailor-make terpene blends for biofuel production. Here, TRV-enabled CRISPR/Cas9 genome editing, including allele-specific knock-out of redundant genes, engineering of enzyme specificity via structure-guided point mutations, and overexpression of terpene genes relevant to stress-protection or biofuel production, would be used to increase metabolic flux toward desired pathways. Broader Impacts - Integrating the system-wide discovery, mechanistic analysis and non-transgenic genome engineering of the switchgrass terpene network aligns the required steps to unlock the chemical potential of this important metabolite class to generate crops that are more resistant to stress and provide advanced biofuel production in light of rising climate pressures as foreseeable challenges for bioenergy crop cultivation. The proposed project would further offer interdisciplinary student training through active involvement in the project and integration of research concepts and outcomes into newly-developed graduate and undergraduate courses on Plant Biotechnology.

09 BIOMASS FUELS↗

Potential for Genetic Improvement of Sugarcane as a Source of Biomass for Biofuels

Sugarcane (Saccharum spp. hybrids) has great potential as a major feedstock for biofuel production worldwide. It is considered among the best options for producing biofuels today due to an exceptional biomass production capacity, high carbohydrate (sugar + fiber) content, and a favorable energy input/output ratio. To maximize the conversion of sugarcane biomass into biofuels, it is imperative to generate improved sugarcane varieties with better biomass degradability. However, unlike many diploid plants, where genetic tools are well developed, biotechnological improvement is hindered in sugarcane by our current limited understanding of the large and complex genome. Therefore, understanding the genetics of the key biofuel traits in sugarcane and optimization of sugarcane biomass composition will advance efficient conversion of sugarcane biomass into fermentable sugars for biofuel production. The large existing phenotypic variation in Saccharum germplasm and the availability of the current genomics technologies will allow biofuel traits to be characterized, the genetic basis of critical differences in biomass composition to be determined, and targets for improvement of sugarcane for biofuels to be established. Emerging options for genetic improvement of sugarcane for the use as a bioenergy crop are reviewed. This will better define the targets for potential genetic manipulation of sugarcane biomass composition for biofuels.

09 BIOMASS FUELS↗

Lessons Learned from US Experience with Biofuels: Comparing the Hype with the Evidence

Biofuel production in the United States, primarily from corn, has more than doubled since 2007, leading to concerns about its unintended consequences for agricultural and fuel markets. To examine the validity of these concerns and inform the debate about biofuels and their impacts, we review ex ante projections and ex post evidence of the effects of biofuels on land use, food and fuel prices, and greenhouse gas (GHG) emissions. We find that biofuels expansion contributed to an initial significant increase in agricultural commodity prices, but these impacts have dissipated over time as crop productivity has increased and cropping patterns have changed. Simulated estimates of indirect land use change and the related GHG emissions intensity of biofuels have also declined sharply from their early levels, which is consistent with ex post evidence. Additionally, growth in biofuel production caused a very modest reduction in fossil fuel prices, implying a small fuel rebound effect. Overall, estimates imply that first-generation biofuels from corn have a lower carbon intensity than gasoline. Lastly, learning by doing, economies of scale, and technological improvements have made biofuels from corn increasingly competitive, reducing the need for subsidies and import tariffs. We conclude with a discussion of the lessons learned from the US biofuels experience.

09 BIOMASS FUELS↗

Center for Advanced Biofuel Systems (CABS) Final Report

One of the great challenges facing current and future generations is how to meet growing energy demands in an environmentally sustainable manner. Renewable energy sources, including wind, geothermal, solar, hydroelectric, and biofuel energy systems, are rapidly being developed as sustainable alternatives to fossil fuels. Biofuels are particularly attractive to the U.S., given its vast agricultural resources. The first generation of biofuel systems was based on fermentation of sugars to produce ethanol, typically from food crops. Subsequent generations of biofuel systems, including those included in the CABS project, will build upon the experiences learned from those early research results and will have improved production efficiencies, reduced environmental impacts and decreased reliance on food crops. Thermodynamic models predict that the next generations of biofuel systems will yield three- to five-fold more recoverable energy products. To address the technological challenges necessary to develop enhanced biofuel systems, greater understanding of the non-equilibrium processes involved in solar energy conversion and the channeling of reduced carbon into biofuel products must be developed. The objective of the proposed Center for Advanced Biofuel Systems (CABS) was to increase the thermodynamic and kinetic efficiency of select plant- and algal-based fuel production systems using rational metabolic engineering approaches grounded in modern systems biology. The overall strategy was to increase the efficiency of solar energy conversion into oils and other specialty biofuel components by channeling metabolic flux toward products using advanced catalysts and sensible design:1) employing novel protein catalysts that increase the thermodynamic and kinetic efficiencies of photosynthesis and oil biosynthesis; 2) engineering metabolic networks to enhance acetyl-CoA production and its channeling towards lipid synthesis; and 3) engineering new metabolic networks for the production of hydrocarbons required to meet commercial fuel standards.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Greenhouse gas emission reduction and cost from the United States biofuels mandate

The United States biofuel mandate might be the largest operating program to reduce greenhouse gas (GHG) emissions. Our paper uses regulatory information about abatement and compliance cost estimates to measure the greenhouse gas abatement cost curve of the mandate. Historical compliance costs per unit of biofuel appear readily available, yet findings here show that using these data along with observed biofuel quantities biases biofuel mandate abatement cost estimates upward. Unbiased estimation requires that related biofuel mandate costs and emissions are estimated along with associated market effects, including on fuel use, using an economic model. Moreover, this approach relates greenhouse gas abatement to the implications for crop prices and land use, taking account of interactions within the crop sector. Results show that the United States biofuel mandate causes up to 20 million metric tons of carbon dioxide equivalent abatement for compliance costs that are equivalent to carbon prices ranging up to $20 per ton, with a large part coming from reductions in petroleum product use as the mandate encourages substitution to biofuels. Abatement costs are reduced if ethanol is made at new facilities with fewer emissions or if petroleum output expansion causes greater emissions than the historical average. Results also indicate that GHG abatement via the mandate is initially costly due to the nature of biofuel demand, there can be a trade-off between corn price increase and GHG reduction, and the increase in total area planted to crops in the U.S. is less important than the reallocation of land among crops.

09 BIOMASS FUELS↗

Dynamic life-cycle carbon analysis for fast pyrolysis biofuel produced from pine residues: implications of carbon temporal effects

Abstract Background Woody biomass has been considered as a promising feedstock for biofuel production via thermochemical conversion technologies such as fast pyrolysis. Extensive Life Cycle Assessment studies have been completed to evaluate the carbon intensity of woody biomass-derived biofuels via fast pyrolysis. However, most studies assumed that woody biomass such as forest residues is a carbon–neutral feedstock like annual crops, despite a distinctive timeframe it takes to grow woody biomass. Besides, few studies have investigated the impacts of forest dynamics and the temporal effects of carbon on the overall carbon intensity of woody-derived biofuels. This study addressed such gaps by developing a life-cycle carbon analysis framework integrating dynamic modeling for forest and biorefinery systems with a time-based discounted Global Warming Potential (GWP) method developed in this work. The framework analyzed dynamic carbon and energy flows of a supply chain for biofuel production from pine residues via fast pyrolysis. Results The mean carbon intensity of biofuel given by Monte Carlo simulation across three pine growth cases ranges from 40.8–41.2 g CO 2 e MJ −1 (static method) to 51.0–65.2 g CO 2 e MJ −1 (using the time-based discounted GWP method) when combusting biochar for energy recovery. If biochar is utilized as soil amendment, the carbon intensity reduces to 19.0–19.7 g CO 2 e MJ −1 (static method) and 29.6–43.4 g CO 2 e MJ −1 in the time-based method. Forest growth and yields (controlled by forest management strategies) show more significant impacts on biofuel carbon intensity when the temporal effect of carbon is taken into consideration. Variation in forest operations and management (e.g., energy consumption of thinning and harvesting), on the other hand, has little impact on the biofuel carbon intensity. Conclusions The carbon temporal effect, particularly the time lag of carbon sequestration during pine growth, has direct impacts on the carbon intensity of biofuels produced from pine residues from a stand-level pine growth and management point of view. The carbon implications are also significantly impacted by the assumptions of biochar end-of-life cases and forest management strategies.

09 BIOMASS FUELS↗

Computational Resources for Biofuel Feedstock Species

While current production of ethanol as a biofuel relies on starch and sugar inputs, it is anticipated that sustainable production of ethanol for biofuel use will utilize lignocellulosic feedstocks. Candidate plant species to be used for lignocellulosic ethanol production include a large number of species within the Grass, Pine and Birch plant families. For these biofuel feedstock species, there are variable amounts of genome sequence resources available, ranging from complete genome sequences (e.g. sorghum, poplar) to transcriptome data sets (e.g. switchgrass, pine). These data sets are not only dispersed in location but also disparate in content. It will be essential to leverage and improve these genomic data sets for the improvement of biofuel feedstock production. The objectives of this project were to provide computational tools and resources for data-mining genome sequence/annotation and large-scale functional genomic datasets available for biofuel feedstock species. We have created a Bioenergy Feedstock Genomics Resource that provides a web-based portal or clearing house for genomic data for plant species relevant to biofuel feedstock production. Sequence data from a total of 54 plant species are included in the Bioenergy Feedstock Genomics Resource including model plant species that permit leveraging of knowledge across taxa to biofuel feedstock species. We have generated additional computational analyses of these data, including uniform annotation, to facilitate genomic approaches to improved biofuel feedstock production. These data have been centralized in the publicly available Bioenergy Feedstock Genomics Resource (http://bfgr.plantbiology.msu.edu/).

09 BIOMASS FUELS↗

Marine Algae Industrialization Consortium (MAGIC): Combining biofuel and high-value bioproducts to meet the RFS

The Marine Algae Industrialization Consortium (MAGIC) was formed to address pressing challenges in the commercialization of microalgae as a source of biofuel. The “Marine Algae Industrialization Consortium (MAGIC): Combining biofuel and high-value bioproducts to meet the RFS” project formally addressed two US Department of Energy Bioenergy Technologies Office (BETO) goals: (1) Model the sustainable supply of 1 million metric tonnes ash free dry weight (AFDW) cultivated algal biomass and (2) Demonstrate valuable co-products produced along with biofuel intermediates to increase value of algal biomass by 30%. To achieve these goals, the project demonstrated and validated high-value co-products to drive down the cost of biofuel by increasing the value of algae “co-products” towards increasing the selling price of total algae biomass as one of the key drivers of economics and adoption. This was accomplished through five core, interdependent tasks including: (1) strain selection to identify and deliver strains for mass culture, (2) mass culture using a hybrid cultivation system and following key operating parameters for downstream applications to provide algae feedstock, (3) recovery and conversion to evaluate two alternative methods to separate dry algae biomass into oil and residuals for downstream testing, (4) product assessment to determine biofuel, aquafeed or poultry feed product efficacy using algae biomass fractions as well as to provide critical performance data for valuation and (5) commercialization to use technoeconomic and life cycle assessments (TEA/LCA) as iterative design and assessment tools including consideration of target markets, competitors, and distribution channels to guide product assessment, development and valuation. A total of 46 peer-review publications, many open-access, provide detail of much of the work carried out and the results of the tasks. Additional reports and presentations provide other technical and public engagement material. At a high level, using a variety of approaches, more than 1000 marine microalgae strains were evaluated to ultimately identify the seven winners that were down-selected to be grown in mass culture. Strain selection demonstrated that there were no ‘super strains’ and that each candidate had strengths and limitations for specific products, growth conditions or operational considerations. Mass culture growth of these seven strains at >5000 L / 29 m 2 scale found that four them were suitable for product assessment. More than 250 kg of biomass was produced across hundreds of pond runs along with thousands of cultivation entries on the growth and biomass characteristics as well as environmental parameters. In the process, dozens of standard operating procedures were generated as was custom software to process and analyze cultivation data. Recovery and conversion of algae biomass demonstrated that a hexane solvent based extraction protocol was most effective at recovering oil (biocrude) from algae and four strains were processed to produce oil and lipid extracted algae (residuals) for downstream testing. Membrane-based oil separation was less successful, but may still be applicable to other commercial applications in the future. Product testing demonstrated that algae biocrude is of high quality and hydrotreating generated numerous fractions of high quality composition for fuel and lubricate based applications. Aquafeed studies performed at a variety of scales showed that both whole and defatted (lipid extracted algae) microalgae were suitable as a feed ingredient, but that the specifics of the fed animal and biochemical composition of the algae are critical factors when determining formulation. Similarly, poultry studies on whole and defatted microalgae generally showed positive outcomes on animal growth and health, with some microalgae providing enhanced nutritional composition of the animal product. Economic and life cycle assessments covered a wide range of possible commercialization and sustainability scenarios. Replacement value, improved product value added, consumer values marketing added valuation and improved animal health were considered as alternatives for microalgae valuation. Using the open pond system, algae productivity was identified as the key driver of commercialization economics, but combination of co-products (e.g. animal feed) with biofuel production substantially increased the total selling price of algae. Modeled microalgae selling price exceeded $\$$1500/tonne and could generate competitive biofuel selling prices below $\$$5 gallon gas equivalents using realistic algal productivities. Short (process scale) and longer (decadal trends) sustainability assessments show that marine microalgae can enhance the sustainability of energy production and lead to other realized benefits in water, fertilizer and land use for other sectors (e.g. agriculture). This project successfully demonstrated all of the components of an end-to-end process from mass microalgae cultivation and dewatering, to recovery and conversion of algae biomass components, to final product demonstration and process valuation; the combined results provide a framework for future commercialization of algae based biofuels.

09 BIOMASS FUELS↗

Future marine biofuels in the port of Seattle region

Marine transportation, a vital global sector, emits 3% of global annual greenhouse gas emissions, which are predicted to increase in the future. Marine biofuels derived from biomass or waste sources like wood residue, waste oil and municipal solid waste can be used for decarbonization. However, limited studies have explored if sufficient marine biofuels could be produced and supplied to major regional ports given feedstock, supply chain and technological constraints. We fill this gap by evaluating the feasibility of supplying marine biofuels to the Port of Seattle. The Regional Bio-Economy Model (RBEM) and the Freight and Fuel Transportation Optimization Tool (FTOT) are used to build scenarios for simulating marine biofuel production in the Port region. We harmonized technoeconomic assumptions for RBEM and FTOT, input FTOT feedstock utilization and routing outputs into RBEM, and modelled conversion, feedstock, and policy scenario variations in RBEM. In RBEM, overall biofuel production was constrained primarily by the biofuel cost, and then by feedstock availability. Providing policy incentives and reducing permitting time frames alleviated these constraints and spurred the buildout of a robust industry through industrial learning dynamics in the initial years. With these measures in place, the RBEM results show that 100% of fuel demand at the Port can be supplied by biofuels with policy incentives and suitable technoeconomic conditions, but the addition of transportation cost considerations using FTOT led to 27.8% of demand being able to be met by biofuels at reasonable fuel delivery cost.

09 BIOMASS FUELS↗

Alternative Crops and Biofuel Production

In order for the biofuel industry to meet the RFS benchmarks for biofuels, new feedstock sources and production systems will have to be identified and evaluated. The Southern Plains has the potential to produce over a billion gallons of biofuels from regionally produced alternative crops, agricultural residues, and animal fats. While information on biofuel conversion processes is available, it is difficult for entrepreneurs, community planners and other interested individuals to determine the feasibility of biofuel processes or to match production alternatives with feed stock availability and community infrastructure. This project facilitates the development of biofuel production from these regionally available feed stocks. Project activities are concentrated in five major areas. The first component focused on demonstrating the supply of biofuel feedstocks. This involves modeling the yield and cost of production of dedicated energy crops at the county level. In 1991 the DOE selected switchgrass as a renewable source to produce transportation fuel after extensive evaluations of many plant species in multiple location (Caddel et al,. 2010). However, data on the yield and cost of production of switchgrass are limited. This deficiency in demonstrating the supply of biofuel feedstocks was addressed by modeling the potential supply and geographic variability of switchgrass yields based on relationship of available switchgrass yields to the yields of other forage crops. This model made it possible to create a database of projected switchgrass yields for five different soil types at the county level. A major advantage of this methodology is that the supply projections can be easily updated as improved varieties of switchgrass are developed and additional yield data becomes available. The modeling techniques are illustrated using the geographic area of Oklahoma. A summary of the regional supply is then provided.

09 BIOMASS FUELS↗