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At least 109 records · Page 6

AlgaeOrtho, a bioinformatics tool for processing ortholog inference results in algae

Introduction: Microalgae constitute a prominent feedstock for producing biofuels and biochemicals by virtue of their prolific reproduction, high bioproduct accumulation, and the ability to grow in brackish and saline water. However, naturally occurring wild type algal strains are rarely optimal for industrial use; therefore, bioengineering of algae is necessary to generate superior performing strains that can address production challenges in industrial settings, particularly the bioenergy and bioproduct sectors. One of the crucial steps in this process is deciding on a bioengineering target: namely, which gene/protein to differentially express. These targets are often orthologs which are defined as genes/proteins originating from a common ancestor in divergent species. Although bioinformatics tools for the identification of protein orthologs already exist, processing the output from such tools is nontrivial, especially for a researcher with little or no bioinformatics experience. Methods: The present study introduces AlgaeOrtho, a user-friendly tool that builds upon the SonicParanoid orthology inference tool (based on an algorithm that identifies potential protein orthologs based on amino acid sequences) and the PhycoCosm database from JGI (Joint Genome Institute) to help researchers identify orthologs of their proteins of interest in multiple diverse algal species. Results: The output of this application includes a table of the putative orthologs of their protein of interest, a heatmap showing sequence similarity (%), and an unrooted tree of the putative protein orthologs. Notably, the tool would be instrumental in identifying novel bioengineering targets in different algal strains, including targets in not-fully annotated algal species, since it does not depend on existing protein annotations. We tested AlgaeOrtho using three case studies, for which orthologs of proteins relevant to bioengineering targets, were identified from diverse algal species, demonstrating its ease of use and utility for bioengineering researchers. Discussion: This tool is unique in the protein ortholog identification space as it can visualize putative orthologs, as desired by the user, across several algal species.

09 BIOMASS FUELS↗

Deploying Purpose-Grown Energy Crops for Sustainable Aviation Fuel (Workshop Summary Report)

From June 6–7, 2023, the U.S. Department of Energy (DOE) Bioenergy Technologies Office (BETO) hosted the workshop “Deploying Purpose-Grown Energy Crops for Sustainable Aviation Fuel” in Kansas City, Missouri. The purpose of the workshop was to examine topics related to large-scale cultivation of purpose-grown energy crops—including algae—that can contribute feedstocks for the production of biofuels and bioproducts. This workshop was a means to gather a variety of stakeholders with the intent to identify elements for a potential future research strategy designed to overcome challenges and delve into the barriers related to the large-scale cultivation of purpose-grown crops. This document provides an overview and summary of the workshop’s presentations, breakout session discussions, and outcomes.

09 BIOMASS FUELS↗

Offshore Membrane Enclosure for Growing Algai (Omega) System for Biofuel Production, Wastewater Treatment, and CO2 Sequestration

We are developing Offshore Membrane Enclosures for Growing Algae (OMEGA). OMEGAs are closed photo-bioreactors constructed of flexible, inexpensive, and durable plastic with small sections of semi-permeable membranes for gas exchange and forward osmosis (FO). Each OMEGA modules is filled with municipal wastewater and provided with CO2 from coastal CO2 sources. The OMEGA modules float just below the surface, and the surrounding seawater provides structural support, temperature control, and mixing for the freshwater algae cultures inside. The salinity gradient from inside to outside drives forward osmosis through the patches of FO membranes. This concentrates nutrients in the wastewater, which enhances algal growth, and slowly dewaters the algae, which facilitates harvesting. The concentrated algal biomass is harvested for producing biofuels and fertilizer. OMEGA system cleans the wastewater released into the surrounding coastal waters and functions as a carbon sequestration system.

Trent, Jonathan↗

Hydrothermal liquefaction of wastewater-grown algae to produce synthetic aviation fuel: A combined experimental study and techno-economic assessment

Large-scale algae farms may someday become a consistent source of biomass feedstock for biofuels. Near-term supplies of algal biomass are available at certain water resource recovery facilities as algae cultivation is used as a method for nutrient recovery from specific effluent streams. Algae grown as a service shifts the value to the service rather than its sole use as a feedstock, which could enable the provision of algal biomass at low to no cost to biofuel producers. Hydrothermal liquefaction (HTL) can readily upgrade wet feedstock slurries, such as algae, to produce a carbon-enriched biocrude. The HTL biocrude can be hydrotreated and distilled, producing a variety of distillate fuels, including synthetic aviation fuel (SAF). We present a pathway, showing the experimental production of SAF from wastewater-grown algae via HTL, along with a techno-economic assessment to identify opportunities for process improvements. Critical quality attributes of the SAF, such as density, viscosity, surface tension, and freeze point, were estimated within the expected fuel experience ranges when compared against petroleum jet fuel. The average minimum fuel selling price of fuels from wastewater-grown algae for breakeven economics was $\$9.04$ per gasoline gallon equivalent (GGE). The sale of co-products such as struvite fertilizers and cement additives can add revenue to reduce the net cost. Ultimately, the selling price is influenced by the scale of the HTL processing facility. Adjusting estimations in the process scale, algae yield, and capital cost estimation can lower the price to $\$6.51$/GGE or raise it to $13.07/GGE.

Biofuels↗

Implications of pond reliability on the techno-economic and life cycle environmental impacts of algal biofuels

Despite extensive research on algal bioproducts, there is limited understanding of how pond contamination affects their economics and environmental impacts. This work compared the costs and environmental impacts of algal biofuels across different pond failure scenarios. Pond failure was simulated by a reliability model based on pond mean-time-to-failure (MTTF). The reliability model was integrated with a process model to analyze the impacts of pond failure on the operations of algal farms and biorefineries. Process model outputs were used for techno-economic analysis and life cycle assessment to determine the minimum fuel selling price (MFSP), global warming potential (GWP), and freshwater consumption impacts of algal biofuels for five MTTF scenarios of 20, 54, 80,120, and 350 days, assuming an average mean-time-to-reset of 7 days. Results show that higher MTTFs reduce the cost and environmental impact of algal biofuels, but with diminishing returns. The average MFSPs for the 20-day, 54-day, and 350-day MTTF scenarios were $\$3.52$, $\$2.54$, and $\$2.10$ per liter of gasoline equivalent, respectively. The GWP for the same scenarios were 131, 96, and 83 g CO 2eq MJ –1 , respectively. This study highlights the significant impact of larger seed trains, required under low MTTFs, on the costs and greenhouse gas emissions of algal biofuels. Moreover, the work shows that algal biofuels fail to be cost-competitive with conventional fuels, even when productivities are increased from 17 to 35 g m –2 d –1 . Furthermore, this work is the first to explore the implications of pond failure on the sustainability of algal biofuels and provides valuable insights to algae farmers on how to reduce the costs and financial risks of algal cultivation through process design and pond management strategies.

09 BIOMASS FUELS↗

Continuous measurements of volatile gases as detection of algae crop health

Wide adoption of algae cultivation to produce environmentally sustainable biofuels and fine chemicals is currently hampered by large losses (10 to 30%) incurred by grazer infections. We show the usage of real-time chemical ionization mass spectrometry to rapidly identify gaseous indicators of grazer infections in cyanobacteria cultures. Grazing was detected significantly faster (up to 3 d) using real-time mass spectrometry than the current methods of microscopy and qPCR. By employing this technology, cultivators may be empowered to treat grazer infestations sooner, thereby protecting the crop and enhancing profitability.

09 BIOMASS FUELS↗

Tailored Bioblendstocks With Low Environmental Impact To Optimize MCCI Engines

The prohect goal is to develop and demonstrate a microalgae bio-blendstock with greater than 60% greenhouse gas reduction potential relative to petroleum diesel, that can reduce sooting propensity, increase cetane number and improve engine thermal efficiency relative to a baseline diesel engine operating on conventional fuel.

09 BIOMASS FUELS↗

Harnessing genetic engineering to drive economic bioproduct production in algae

Our reliance on agriculture for sustenance, healthcare, and resources has been essential since the dawn of civilization. However, traditional agricultural practices are no longer adequate to meet the demands of a burgeoning population amidst climate-driven agricultural challenges. Microalgae emerge as a beacon of hope, offering a sustainable and renewable source of food, animal feed, and energy. Their rapid growth rates, adaptability to non-arable land and non-potable water, and diverse bioproduct range, encompassing biofuels and nutraceuticals, position them as a cornerstone of future resource management. Furthermore, microalgae’s ability to capture carbon aligns with environmental conservation goals. While microalgae offers significant benefits, obstacles in cost-effective biomass production persist, which curtails broader application. This review examines microalgae compared to other host platforms, highlighting current innovative approaches aimed at overcoming existing barriers. These approaches include a range of techniques, from gene editing, synthetic promoters, and mutagenesis to selective breeding and metabolic engineering through transcription factors.

09 BIOMASS FUELS↗

Low Molecular Weight Volatile Organic Compounds Indicate Grazing by the Marine Rotifer Brachionus plicatilis on the Microalgae Microchloropsis salina

Microalgae produce specific chemicals indicative of stress and/or death. The aim of this study was to perform non-destructive monitoring of algal culture systems, in the presence and absence of grazers, to identify potential biomarkers of incipient pond crashes. Here, we report ten volatile organic compounds (VOCs) that are robustly generated by the marine alga, Microchloropsis salina, in the presence and/or absence of the marine grazer, Brachionus plicatilis. We cultured M. salina with and without B. plicatilis and collected in situ volatile headspace samples using thermal desorption tubes over the course of several days. Data from four experiments were aggregated, deconvoluted, and chromatographically aligned to determine VOCs with tentative identifications made via mass spectral library matching. VOCs generated by algae in the presence of actively grazing rotifers were confirmed via pure analytical standards to be pentane, 3-pentanone, 3-methylhexane, and 2-methylfuran. Six other VOCs were less specifically associated with grazing but were still commonly observed between the four replicate experiments. Through this work, we identified four biomarkers of rotifer grazing that indicate algal stress/death. This will aid machine learning algorithms to chemically define and diagnose algal mass production cultures and save algae cultures from imminent crash to make biofuel an alternative energy possibility.

headspace sampling↗

Synthetic and Biomass Alternate Fueling in Aviation

Worldwide, aviation alone uses 85 to 95 billion gallons of nonrenewable fossil fuel per year (2008). General transportation fueling can accommodate several different fuels; however, aviation fuels have very specific requirements. Biofuels have been flight demonstrated, are considered renewable, have the capacity to become "drop-in" replacements for Jet-A fuel, and solve the CO2 climate change problem. The major issue is cost; current biomass biofuels are not economically competitive. Biofuel feedstock sources being researched are halophytes, algae, cyanobacteria, weeds-to-crops, wastes with contingent restraints on use of crop land, freshwater, and climate change. There are five major renewable energy sources: solar thermal, solar photovoltaic, wind, drilled geothermal and biomass, each of which have an order of magnitude greater capacity to meet all energy needs. All five address aspects of climate change; biomass has massive potential as an energy fuel feedstock.

Hendricks, R.C.↗

Biocontainment of Genetically Engineered Algae

Algae (including eukaryotic microalgae and cyanobacteria) have been genetically engineered to convert light and carbon dioxide to many industrially and commercially relevant chemicals including biofuels, materials, and nutritional products. At industrial scale, genetically engineered algae may be cultivated outdoors in open ponds or in closed photobioreactors. In either case, industry would need to address a potential risk of the release of the engineered algae into the natural environment, resulting in potential negative impacts to the environment. Genetic biocontainment strategies are therefore under development to reduce the probability that these engineered bacteria can survive outside of the laboratory or industrial setting. These include active strategies that aim to kill the escaped cells by expression of toxic proteins, and passive strategies that use knockouts of native genes to reduce fitness outside of the controlled environment of labs and industrial cultivation systems. Several biocontainment strategies have demonstrated escape frequencies below detection limits. However, they have typically done so in carefully controlled experiments which may fail to capture mechanisms of escape that may arise in the more complex natural environment. The selection of biocontainment strategies that can effectively kill cells outside the lab, while maintaining maximum productivity inside the lab and without the need for relatively expensive chemicals will benefit from further attention.

59 BASIC BIOLOGICAL SCIENCES↗

An Algal Nutrient‐Replete, Optimized Medium for Fast Growth and High Triacylglycerol Accumulation

Microalgae are promising sources to sustainably meet the global needs for energy and products. Algae grow under different trophic conditions, where nutritional status regulates biosynthetic pathways, energy production, and growth. The green alga Chromochloris zofingiensis has strong economic potential because it co-produces biofuel precursors and the high-value antioxidant astaxanthin while accumulating biomass when grown mixotrophically. As an emerging reference alga for photosynthesis, metabolism, and bioproduction, C. zofingiensis needs a defined, optimized medium to standardize experiments during fast growth for batch cultivation. Because the interplay of glucose treatment (+Glc) and mineral deficiency influences photosynthesis, growth, and the production of lipids and astaxanthin, we designed a replete nutrient medium tailored to the C. zofingiensis cellular ionome. We combined inductively coupled plasma mass spectrometry (ICP-MS) and +Glc growth curves to determine a medium that is nutrient replete for at least 5 days of +Glc logarithmic growth. We found that there are high nutritional needs for phosphorus and sulfur during mixotrophy. Iron was the only element measured for which the cellular concentration correlated with exogenous concentration and was iteratively adjusted until the cellular ionome was consistent through the logarithmic growth phase. This Chromochloris-Optimized Ratio of Elements (CORE) medium supports fast growth and high biomass and lipid accumulation without causing excess nutrient toxicity. This defined, nutrient-replete standard is important for future C. zofingiensis investigations and can be adapted for other species to support high biomass for batch cultivation. The method used to develop CORE medium shows how ionomics informs replicable media design and may be applied in industrial settings to inform cost-effective biofuel production.

Jeffers, Tim L. [University of California, Berkele↗

Feasibility of Algal Biochar, a Byproduct of Biofuel Production, as a Supplemental Cementitious Material

Algal biochar, as the solid residue of biofuel production from algal biomass, is reported to explore disposition options, aiming to lessen the liability or obstacles to biofuel production processes. However, landfills and open combustion lead to adverse environmental impacts. One way to add value to such wastes is to use them as admixtures in cementitious construction materials. This study aims to investigate the feasibility of algae-derived biochar as supplementary cementitious materials (SCM) at different water contents and mixture ratios. Algal biochar-cement composites were prepared with different algal biochar content as well as different water-to-cement (w/c) ratios, and the surface area, morphology, elemental, and mineralogical composition were characterized. To compensate for the high-water absorption of algal biochar, a small concentration of a superplasticizer was used since higher w/c ratios negatively impact strength. The mechanical performance of algal biochar-cement composites is compared with control composites using commercial silica fume as a typical commercial SCM. The findings suggest that algal biochar is a promising candidate to replace commercial SCM, like silica fume, since algal biochar-cement composites can reach comparable compressive strength and Young’s modulus to commercial pozzolan-cement materials with the same w/c ratio, though at later curing times, 33 days. Although the tensile strength of algal biochar-cement composites is statistically similar at 7 days, it is significantly lower at later curing times, and further investigation is required to improve this property. Algal biochar-based cement binders showed comparable embodied carbon to silica fume-based cement binders based on a cradle-to-gate lifecycle analysis. However, the ability of algal biochar to absorb large volumes of CO 2 over short periods of time, as measured in this study, makes this novel SCM an excellent alternative to reduce the embodied carbon of concrete structures cradle-to-grave at 1/10 of the cost. In conclusion, valorization of algae-derived solid waste provides great potential to reduce embodied carbon and brings credit to biofuel production and concrete-based construction.

algae-derived↗

Biodiversity and disease risk in an algal biofuel system: An experimental test in outdoor ponds using a before-after-control-impact (BACI) design

For outdoor cultivation of algal feedstocks to become a commercially viable and sustainable option for biofuel production, algal cultivation must maintain high yields and temporal stability in environmentally variable outdoor ponds. One of the main challenges is mitigating disease outbreaks that leads to culture crashes. Drawing on predictions from the ‘dilution effect’ hypothesis, in which increased biodiversity is thought to reduce disease risk in a community, a teste of whether algal polycultures would reduce disease risk and improve feedstock production efficiencies compared to monocultures was performed. While the positive benefits of biodiversity on disease risk have been demonstrated in various systems, to the best of our knowledge this is the first test in an algal biofuel system. Here, the results a before-after-control-impact (BACI) experimental design to compare mean monoculture (control) and polyculture (impact) yield, stability, and productivity before and after fungal infection when grown in 400-L outdoor raceway ponds are presented. It has been found that polycultures did not experience a reduction in disease risk compared to monocultures or differ in production efficiencies throughout the course of the 43-day experiment. These results show that polyculture feedstocks can maintain similar levels of productivity, stability, and disease resistance to that of a monoculture. Determining whether these results are generalizable or represent one case study requires additional outdoor experiments using a larger variety of host and pathogen species.

09 BIOMASS FUELS↗

A Novel Platform for Algal Biomass Production Using Cellulosic Mixotrophy (CeMix) (Final Technical Report)

This novel project, branded CeMIX for cellulosic mixotrophy, targets solutions for several technological barriers limiting the deployment of algae cultivation in sunny, arid regions (specifically the southwest United States). Opening up the region for algae cultivation could increase DOE’s resource assessments and biofuel potential for the U.S. Mixotrophic metabolism in support of liquid fuel production is completely dependent on the availability of waste organic carbon in order to avoid diversion of food resources for fuel production. The use of cellulosic sugar hydrolysate (CSH) produced via the NREL process provided a standardized carbon source for study of the underlying biochemistry and metabolic adjustments to mixotrophy. Mixotrophic metabolism uses both photosynthetic CO 2 fixation and sugar oxidation to improve biomass productivity and harvest yield. The project was high-risk, high-reward in the sense that adding organic carbon to algal cultivation is an invitation for rapid-growth, heterotrophic contaminants to overtake the culture and consume mineral nutrients needed for algal growth. To help evade this outcome the project focused on acidophilic red algae that require low pH conditions with 40-48°C optimal temperature profiles that define them as low-range thermophiles. This is the only project in the BETO portfolio to utilize extreme conditions of low pH and high operating temperatures to control pathogens and competitors in mass culture. The project yielded five major outcomes. 1) Harvest densities of 5-10 g ash-free dry weight are easily achieved, providing a 10-fold reduction in water required and dewatering demand. 2) Catabolic repression of photosynthesis in G. sulphuraria is conditionally repressed by low oxygen. 3) Mixotrophic growth on cellulosic hydrolysate consumes all C6 and C5 sugars concomitant with increases in floridean starch with little change in lipid content. Protein content can be manipulated between 35 and 54 wt% by varying the C:N molar ratio from 10:1 to 20:1, respectively. 4) Mixotrophic cultures with up to 50 mM total sugar are remarkably stable at pH 2.5 and daily average reactor temperatures of 40°C. 5) Capital costs for glass tubular photobioreactors are prohibitive for liquid fuel production using a hydrothermal liquefaction processing pathway. The economic outlook for covered raceway ponds is better with a project minimum fuel selling price at $3.32 dm 3 GE -1 .

09 BIOMASS FUELS↗

Air Carbon for Algae Production (AirCAP) – Expanding algae resource potential via direct (in-pond) air-CO2 capture.

Microalgae biofuel production potential in the United States is constrained by a lack of suitable cultivation locations within a close proximity to CO2 point-sources. Initial estimates suggest that removing the requirement of a concentrated CO2 source expands resource potential ten-fold, allowing the projection of production levels that approach levels of current U.S. diesel consumption. Decoupling cultivation locations from concentrated CO2 point-sources requires direct capture of air-CO2 into the pond bulk at a rate sufficient to meet maximum expected carbon assimilation rates (10 g C/m2-day and higher). Enhancement of the ingassing rate via chemical reaction, the direct reaction of dissolved CO2 with hydroxide ions, has been proposed as a mechanism to increase the ingassing rate of air-CO2. The central objectives of this project are to validate whether chemically enhanced air-CO2 ingassing rates in large raceway ponds can meet or exceed the level required to support high levels of algae productivity, then subsequently identify alkaliphilic strains able to withstand such conditions while maintaining a biomass productivity that enables process economics. Full scale ponds (~1 acre, unlined) will be used to validate ingassing rates as a function of pH expected at scale in a series of abiotic experiments. To date, mass-transfer dynamics have been characterized in ponds ranging from 1 to 1,000 m2. Initial results will be presented describing the mass transfer coefficient in 1-acre (~4,000 m2), unlined ponds. The maximum expected carbon ingassing rate will be measured during ingassing trials, in which pond pH is displaced from equilibrium via the addition of a strong base, with the subsequent decrease in pH (and increase in dissolved inorganic carbon) corresponding to the air-CO2 absorption rate. A mass-transfer model, validated with experimental results, will be used to project ingassing rates expected at pond scales (10 acres) required for production of commodity chemicals.

09 BIOMASS FUELS↗

Improving the Economic Viability of Biological Utilization of Coal Power Plant CO 2 by Improved Algae Productivity and Integration with Wastewater Treatment

The overall goal of this project was to improve the cost-effectiveness of systems using algae to capture and beneficially utilize CO 2 from the flue gas of coal-fired power plants. Algal biomass cultivation is an attractive alternative for carbon capture and utilization because it can uptake more CO 2 per land area than any other photosynthetic crop. Also, the resulting biomass can be used to make valuable large-market commodity products, such as biofuels and animal feed. However, the use of algal carbon capture has been limited by several factors including: (1) high costs for growing algae (>$\$1,300$/ton), (2) large land requirements for algae cultivation, and (3) environmental concerns about the large amount of water and nutrient inputs needed for growing algae. This project has taken direct aim at resolving these key limitations and demonstrated several viable approaches to improve the economic and environmental feasibility of algal carbon capture. In particular, this project demonstrated a significant increase in average annual algal productivity from baseline values of 8 g/m 2 -day to above 20 g/m 2 -day. This supports a much lower projected cost of algae production (<$\$500$/ton) and reduces land requirements by more than 50%. Additionally, this project confirmed the ability to grow multiple algae species with wastewater nutrients, which facilitates the potential for substantial new revenue streams for algae cultivation based on tipping fees for nutrient laden wastewater. This study highlighted that there is more potential value in wastewater nutrient treatment credits than in the primary commodity products made from algae. The use of wastewater inputs also mitigates the concerns associated with the large amount of water needed for growing algae. The algal biomass grown for this project using wastewater and simulated flue gas inputs were characterized for potential use as an animal feed ingredient. The algae had a protein content in the range of 35-40%, which is between some common commodity animal feed products— distiller’s grains (~25-30% protein) and soybean meal (45-50% protein). In-vitro digestion tests using cattle rumen fluids, showed that this algae biomass was best suited for use as “bypass protein”, which is digested in the animal intestine. Based on the protein content and amino acid profile of the algae grown in this study, it was estimated to have market value in the range of $\$190$-$\$280$/ton. There is potential for higher value feed products if the algae content of omega-3 fatty acids or antioxidants is accounted for. This project also did small scale demonstration of forward osmosis algae dewatering and nanofiltration of hydrothermal liquefaction aqueous products, which were shown to improve the cost and energy balance for producing algal products. When all the improvements demonstrated in this study were included in techno-economic analysis, it showed there was a viable pathway to profitable algal CO 2 utilization with a payback period in the range of six to ten years.

01 COAL, LIGNITE, AND PEAT↗