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Genome assembly and annotation of the biofuel- and bioproduct-relevant microalga Nannochloropsis oceanica CCAP849/10
Nannochloropsis salina is a halotolerant, high-lipid-producing microalga that is being explored as a biofuel production species. Here, we report an improved high-quality draft assembly and annotation for the nuclear genome of N. salina strain CCMP1776.
A draft genome of 906 scaffolds of 115.8 Mb was created for Desmodesmus armatus , a diploid, lipid- and storage carbohydrate-accumulating microalga proven relevant for large-scale, outdoor cultivation, and serves as a model alga platform for improving photosynthetic efficiency and carbon assimilation for next-generation bioenergy production.
While photosynthetic microalgae, such as Chlorella, serve as feedstocks for nutritional oils and biofuels, heterotrophic cultivation can augment growth rates, support high cell densities, and increase triacylglycerol (TAG) lipid content. However, these species differ significantly in their photoautotrophic and heterotrophic characteristics. In this study, the phylogeny of thirty Chlorella strains was determined in order to inform bioprospecting efforts and detailed physiological assessment of three species. The growth kinetics and lipid biochemistry of C. protothecoides UTEX 411, C. vulgaris UTEX 265, and C. sorokiniana UTEX 1230 were quantified during photoautotrophy in Bold's basal medium (BBM) and heterotrophy in BBM supplemented with glucose (10 g L –1 ). Heterotrophic growth rates of UTEX 411, 265, and 1230 were found to be 1.5-, 3.7-, and 5-fold higher than their respective autotrophic rates. With a rapid nine-hour heterotrophic doubling time, Chlorella sorokiniana UTEX 1230 maximally accumulated 39% total lipids by dry weight during heterotrophy compared to 18% autotrophically. Furthermore, the discrete fatty acid composition of each strain was examined in order to elucidate lipid accumulation patterns under the two trophic conditions. In both modes of growth, UTEX 411 and 265 produced 18:1 as the principal fatty acid while UTEX 1230 exhibited a 2.5-fold enrichment in 18:2 relative to 18:1. Although the total lipid content was highest in UTEX 411 during heterotrophy, UTEX 1230 demonstrated a two-fold increase in its heterotrophic TAG fraction at a rate of 28.9 mg L –1 d –1 to reach 22% of the biomass, corresponding to as much as 90% of its total lipids. Interestingly, UTEX 1230 growth was restricted during mixotrophy and its TAG production rate was suppressed to 18.2 mg L –1 d –1 . This constraint on carbon flow raises intriguing questions about the impact of sugar and light on the metabolic regulation of microalgal lipid biosynthesis.
Rotating algae biofilm reactors (RABRs) can reduce energy requirements for wastewater reclamation but require further optimization for implementation at water resource recovery facilities (WRRF). Optimizing RABR operation is challenging because conditions at WRRF change frequently, and disregarding interaction terms related to these changes can produce incorrect conclusions about RABR behavior. This study evaluated the two-way interaction and main effects of four factors on the biomass productivity and phosphorus removal efficiency of a microalgae-bacteria biofilm grown in municipal anaerobic digester centrate, with factor levels and operating conditions selected to mimic a pilot RABR at a WRRF in Utah. Two-way interactions harvesting period*light intensity (LI), harvesting period*temperature, and LI*hydraulic retention time (HRT) had significant effects on biomass productivity: at high temperature and low LI, highest biomass productivity was achieved with a 14-day harvesting period, but at medium temperature and high LI, highest biomass productivity was achieved with a 7-day harvesting period. At high HRT, highest biomass productivity occurred at low LI, but at low HRT, highest biomass productivity occurred at high LI. Phosphorus removal was strongly influenced by LI and occurred most rapidly during the first 2 days HRT, which suggests precipitation contributed significantly to phosphorus removal. These observations provide insight for further RABR optimization.
This project sought to address the technical and economic barriers to carbon dioxide capture and utilization using microalgae. Operating data were collected in 2016 and 2017 during cultivation of Scenedesmus acutus at Duke Energy’s East Bend Station – a coal-fired power plant located in northern Kentucky – using flue gas as the CO 2 source. Algae were grown in a 1200 L “cyclic flow” photobioreactor (PBR) designed by the University of Kentucky. A key finding was that the harvested algae contained only very low concentrations of heavy metals (As, Cd, Hg, Se), consistent with heavy metals incorporation from the supplied nutrients. This indicates that algal biomass produced from coal-derived flue gas would be suitable for a variety of applications, including the production of bioplastics, use as fertilizer, etc. A lifecycle assessment showed that the UK-designed PBR employed in this work qualifies as a net CO 2 capture technology. Indeed, over a 30-year period, net CO 2 capture would equate to 43% of the targeted amount, i.e., the amount captured from the supplied flue gas. A techno-economic analysis indicated that the minimum production cost of Scenedesmus acutus biomass in the US is in the order of 875 USD/ton, excluding the cost of capital. While this figure is not too dissimilar to values reported for open raceway ponds in similar scenarios, it emphasizes that for current cultivation technology any pathway to economic viability will require applications for which algal boimass can be sold at prices in excess of 1,000 USD/ton. Currently, such applications represent relatively small markets, such as pigments (e.g., astaxanthin) and nutraceuticals (ω-3 unsaturated fatty acids), as well as nutritional supplements (whole algae) for human consumption and for use in pet food. Consequently, the commercialization of large-scale algae-based CO 2 capture and utilization will require the development of new technologies to reduce the cost of algae production and/or the development of new, high-value applications for algal biomass. One of the more promising applications for algal biomass is in the production of bioplastics. In this work, the potential for the algae grown in the East Bend PBR for the production of bioplastic with adequate mechanical properties was clearly shown. Positive features of the produced biomass included a high protein content and a composition that was generally more homogeneous than biomass grown in open ponds (in which many species may be present). The best candidate for further review, after incorporation into ethylene-vinyl acetate, was a lipid and sugar extracted material, which showed the highest extension values with comparable load values to other UK-derived samples. It also demonstrated extension benefits against the Algix Bloom product (currently offered commercially), even though it contained agglomerates which generally exert a negative effect on mechanical properties. This leads to speculation that with enhanced milling that exists on the commercial scale, the sugar and fat extracted product may be even more competitive. This, in turn, points to the need for additional work in order to assess the properties of such optimized algae-based plastics and the price point they can command.
A preliminary techno-economic analysis (TEA) was developed for the fiscal year 2020 state of technology (SOT) assessment to evaluate the benefits and risks for a large-scale microalga hydrothermal liquefaction (HTL) system based on most recent testing results. The focus of the study is directed toward the conversion system, which consists of five processes: two-stage sequential HTL (SEQHTL), biocrude upgrading to final fuels, bioprocessing for co-product generation, hydrogen generation, and steam cycle. In this system, algae biomass with corn stover supplement during the lower algae productivity seasons (winter, fall, and spring) to match the maximum algae seasonal production rate in summer is employed to maintain a constant plant capacity in all the seasons. Algae only (summer season) or algae/corn stover blended feedstock (other seasons) are sent to a two-stage SEQHTL process. In stage I, the carbohydrates in the feedstock are extracted and separated from the residual solid. The residual solid from stage I is further converted to biocrude in the SEQHTL stage II step. The biocrude is upgraded to final fuel products in an upgrading process. The extract stream from HTL stage I is sent to the bioprocessing section for co-product generation via fermentation of carbohydrate. Lactic acid (LA) is assumed to be the co-product based on current bioprocessing testing results.
The fiscal year (FY) 2021 State of Technology (SOT) Assessment for the hydrothermal liquefaction (HTL) of wastewater (WW)-grown microalgae and biocrude upgrading system was completed and reported here. An industrial partner, Gross-Wen Technologies (GWT), provided algae feedstock cultivated on a revolving algal biofilm (RAB) system by using the primary effluent from a water resource recovery facility (WRRF). This provided algae was tested at PNNL for HTL processing. The experimental results provided the major design basis of the HTL process of the SOT baseline case. The primary effluent of the Metropolitan Water Reclamation District (MWRD) of Greater Chicago was assumed to be the nutrients source for algae growth and the algae yield data per gallon wastewater provided by GWT were used to estimate the total algae production rate and thus the HTL conversion plant scale. Considering different cultivation technologies and wastewater streams with different flow rates and nutrients contents can be used to produce algae, this SOT assessment just provided an example case study for WW-grown algae based HTL conversion to fuels systems. A preliminary economic analysis was developed based on process simulation results. Sensitivity analysis was implemented to evaluate cost impacts of plant scales, potential cost improvements and other key factors.
This report presents an updated "harmonization study" documenting the collaborative analysis of microalgae cultivation and conversion to fuels and products. Four national laboratory modeling teams reconvened to investigate the resource, economic, and environmental sustainability implications of integrated systems encompassing large-scale algae farms and conversion biorefineries. Relative to prior harmonization analyses conducted by these partners, the present effort focuses on more near-term deployment potential based on the use of nutrient-replete, high-protein algal biomass compositions (more readily achievable today without sacrificing cultivation productivity), though this also incurs challenges in lower fuel yields and accordingly higher costs and carbon intensities for the overall integrated systems. Additionally, the present assessment adds further granularity around carbon dioxide (CO 2 ) sourcing and transport via carbon capture of nearby point sources, as well as handling of high-saline cultivation media and resultant blowdown/disposal processing. Finally, this assessment focuses on conversion opportunities to produce both fuel (prioritizing sustainable aviation fuel, in this case via hydrothermal liquefaction) and protein products for the food and feed markets, recognizing growing needs for such products.
This report presents an updated “harmonization study” documenting the collaborative analysis of saline microalgae cultivation and conversion to fuels and products. Four national laboratory modeling teams reconvened to investigate the resource, economic, and environmental sustainability implications of integrated systems encompassing large-scale algae farms and conversion biorefineries. Relative to prior harmonization analyses conducted by these partners, the present effort focuses on more near-term technology potential based on the use of nutrient-replete, high-protein algal biomass compositions (more readily achievable today without sacrificing cultivation productivity) coupled with individual algae farms varying in size but generally smaller at 3,900 acres on average (more realistic in practice than a fixed 5,000-acre farm scale previously considered). Additionally, the present assessment adds further granularity around carbon dioxide (CO 2 ) sourcing and transport via carbon capture of nearby point sources, as well as handling of high-saline cultivation media and resultant blowdown/disposal processing. Finally, this assessment focuses on conversion opportunities to produce both fuel (prioritizing sustainable aviation fuel [SAF], in this case via hydrothermal liquefaction) and protein products for the food and feed markets, recognizing growing needs for such products.
Recent progress towards the implementation of renewable bioenergy production has included microalgae, which have potential to significantly contribute to a viable future bioeconomy. In a current challenging energy landscape, where an increased demand for renewable fuels is projected and accompanied by plummeting fossil fuels' prices, economical production of algae-based fuels becomes more challenging. However, in the context of mitigating carbon emissions with the potential of algae to assimilate large quantities of CO2, there is a route to drive carbon sequestration and utilization to support a sustainable and secure global energy future. This chapter places international energy policy in the context of the current and projected energy landscape. The contribution that algae can make, is summarized as both a conceptual contribution as well as an overview of the commercial infrastructure installed globally. Some of the major recent developments and crucial technology innovations are the results of global government support for the development of algae-based bioenergy, biofuels and bioproduct applications, which have been awarded as public private partnerships and are summarized in this chapter.
Davis, R., A. Coleman, A. Badgett, et al. 2024. “Chapter 7: Emerging Resources: Microalgae, Macroalgae, and Point-Source Carbon Dioxide Waste Streams.” In 2023 Billion‐Ton Report. M. H. Langholtz (Lead). Oak Ridge, TN: Oak Ridge National Laboratory. doi:10.23720/BT2023/2316174.
Microalgae are a potential means of recycling CO2 from industrial point sources. With this in mind, a novel photobioreactor (PBR) was designed and deployed at a coal-fired power plant. To ascertain the feasibility of using waste heat from the power plant to heat algae cultures during cold periods, two heat transfer models were constructed to quantify PBR cooling times. The first, which was based on tabulated data, material properties and the physical orientation of the PBR tubes, yielded a range of heat transfer coefficients of 19–64 W m−2 K−1 for the PBR at wind speeds of 1–10 m s−1. The second model was based on data collected from the PBR and gave an overall heat transfer coefficient of 24.8 W m−2 K−1. Energy penalties associated with waste heat utilization were found to incur an 18%–103% increase in energy consumption, resulting in a 22%–70% reduction in CO2 capture for the scenarios considered. A techno-economic analysis showed that the cost of heat integration equipment increased capital expenditures (CAPEX) by a factor of nine and increased biomass production costs by a factor of three. Although the scenario is thermodynamically feasible, the increase in CAPEX incurs an increase in biomass production cost that is economically untenable.
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.
Climate, energy, and food security are three of the greatest challenges society faces this century. Solutions for mitigating the effects of climate change often conflict with solutions for ensuring society’s future energy and food requirements. For example, BioEnergy with Carbon Capture and Storage (BECCS) has been proposed as an important method for achieving negative CO2 emissions later this century while simultaneously producing renewable energy on a global scale. However, BECCS has many negative environmental consequences for land, nutrient, and water use as well as biodiversity and food production. In contrast, large-scale industrial cultivation of marine microalgae can provide society with a more environmentally favorable approach for meeting the climate goals agreed to at the 2015 Paris Climate Conference, producing the liquid hydrocarbon fuels required by the global transportation sector, and supplying much of the protein necessary to feed a global population approaching 10 billion people.
Disclosed herein are a suite of genetic tools suitable for engineering both the nucleus and chloroplast in diverse microalgae.
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