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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Multi-pronged approach to improving carbon utilization by cyanobacterial cultures

The goal of this project was to increase the efficiency of carbon utilization by a Synechocystis biofuel production strain in a photobioreactor system by at least 50% using a multi-pronged approach that combined the best features of both biological and physicochemical CO 2 capture technologies to: 1) increase rates and extent of CO 2 absorption into the culture medium by addition of biocompatible amine solvents and a nanobubble gas delivery system, and 2) increase cellular rates of inorganic carbon uptake and carbon fixation through genetic engineering. In addition, the project plan included evaluating the use of fermentation effluent gases as a CO 2 source and demonstrating performance under outdoor conditions in closed-pond photobioreactor systems. These combined efforts would be guided by integrated technoeconomic and life-cycle analyses. Both physicochemical CO 2 capture technologies explored proved to be successful. The amine solvent monoethanolamine (MEA) was found to be biocompatible with a Synechocystis laurate production strain. Using an adaptive evolution strategy, an MEA-tolerant laurate production strain was developed that showed significantly increased carbon utilization over the parent strain. To our knowledge this was the first time nanobubble (NB) technology had been used as a carbon delivery system for algae production and it also proved quite successful as a proof of concept. Solutions containing CO 2 nanobubbles (Nano-BG11) at concentrations significantly higher than the aqueous CO 2 concentrations achieved with gas sparging were successfully generated and characterized with regard to bubble size (nm), concentration (NB/L), and pH stability as per project plan. Growth studies proved challenging. However, once the source of a puzzling growth defect was found and remedied Synechocystis was able to grow somewhat better in Nano-BG11 than in standard BG11 medium.

09 BIOMASS FUELS↗
DOE OSTI2020

A long-term pilot-scale algal cultivation on power plant flue gas – Cultivation stability and biomass accumulation

A long-term culture of a selected freshwater microalgal assemblage on power plant flue gas was realized using a pilot-scale photobioreactor. Different culture conditions of light cycle, harvesting amount, pH stabilization, and sulfur addition were tested during a 12-month continuous operation at a natural gas fired power plant. The results concluded that the daily harvesting amount of 60% with no light-cycle and NH 4 NO 3 as the nitrogen source and pH stabilization agent, resulted in optimal cultivation performance. Biomass concentration and biomass productivity reached 1.07 ± 0.23 g L -1 and 0.43 ± 0.14 g L -1 d -1 , respectively. During the year-long cultivation period, the algal assemblage was relatively stable with average abundances of 88 ± 4% and 12 ± 4% for algae and bacteria, respectively, and no significant differences between individual case scenarios. The stable culture fostered a stable biomass composition. Protein, carbohydrate, and lipid contents of the algal biomass were 58.60 ± 2.41%, 19.5 ± 2.44%, and 9.70 ± 1.68%, respectively.

09 BIOMASS FUELS↗
DOE OSTI2021

A validated thermal and biological model for predicting algal productivity in large scale outdoor cultivation systems

Proper assessment of the sustainability of algal products requires an understanding of algal growth rate with regional and temporal resolution. This study developed a bulk growth model that utilizes geospatial data, reactor geometry inputs, and a maximum of six strain-specific parameters to predict algal productivity with temporal and regional resolution. The model requires local weather data to calculate the time-resolved culture temperature which is combined with a biological model based on maximum theoretical carbon fixation to predict the time-resolved algal concentration. Validation of the model against experimental data illustrates an average ac- curacy of 0.27% ± 5.32% when comparing algal concentration at harvest over 45 days of cultivation data for Galdieria sulphuraria 5587.1, and Galdieria sulphuraria Soos grown in photobioreactors (PBRs) in Mesa, AZ (33.4152° N, 111.8315° W). For open raceway ponds (ORPs), an accuracy of 0.9% ± 2.35% was achieved when comparing algal concentration at harvest over 50 days of cultivation data for Chlorella vulgaris, Desmodesmus intermedius, Nannochloropsis oceanica grown in Atlanta, GA (33.7490° N, 84.3880° W) and Mesa, AZ (33.4152° N, 111.8315° W). The validation trials spanned three seasons (winter, spring, and summer) to ensure model ac- curacy throughout the year, and concentration between harvests was modeled with hourly resolution. The validated model is used to assess the productivity at several case-study locations in the United States with Chlorella vulgaris, grown in open raceway ponds and Galdieria sulphuraria grown in vertical flat panel photo- bioreactors. In conclusion, the model has been published as an open-source tool, in an effort to increase the fidelity of future studies that rely on outdoor algae cultivation for feedstock production and allow for time resolved results for future techno-economic and life cycle assessments.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

A streamlined approach to characterize microalgae strains for biomass productivity under dynamic climate simulation conditions

Screening microalgae strains under static light and temperature flask conditions cannot directly quantify the biomass productivities of algae growing in the dynamically fluctuating light and temperature conditions of outdoor ponds. In this effort, we describe a testing pipeline that screens for productivity under climate simulated conditions. A validated, miniaturized Laboratory Environmental Algae Pond Simulator (mini-LEAPS) photobioreactor was used to determine optimal medium salinities and biomass productivities of cold-tolerant microalgae collected from Arctic, subarctic, Antarctic, and subalpine habitats. Strains characterized in this effort include: Chlorella antarctica UTEX1959, Chlorella sp. UTEXSNO69, Chloromonas rosae UTEXSNO11, Phaeodactylum tricornutum UTEX646, and Stichococcus minutus CCALA727. Observed productivities of the characterized strains ranged from 1.74 ± 0.25 to 8.18 ± 0.81 g m –2 day –1 (as ash-free dry weight). For each strain, a temperature tolerance profile was generated to identify the most appropriate season (s) for cultivation. The two most promising strains, Chlorella sp. UTEX SNO69 and P. tricornutum UTEX646, were tested alongside the DISCOVR winter benchmark strain Monoraphidium minutum 26B-AM in outdoor open ponds at the PNNL Algae Testbed (PAT) in Arizona. Further, under cold-season outdoor pond conditions, Chlorella sp. UTEXSNO69 achieved average areal biomass productivities of 8.21 ± 0.50 g m –2 day –1 , not significantly different from that of the benchmark strain (8.52 ± 0.43 g m –2 day –1 , p > 0.05). Under spring outdoor pond conditions, P. tricornutum UTEX646 achieved average areal biomass productivities 10.34 ± 0.27 g m –2 day –1 , not significantly different from that of the benchmark (10.07 ± 0.50 g m –2 day –1 ) under the tested conditions (p > 0.05). The streamlined pipeline was thus demonstrated to successfully characterize productive microalgae strains for outdoor deployment by weeding out algae strains that were not productive under the tested dynamic light and temperature conditions.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Community structure and function during periods of high performance and system upset in a full-scale mixed microalgal wastewater resource recovery facility

Microalgae have the potential to exceed current nutrient recovery limits from wastewater, enabling water resource recovery facilities (WRRFs) to achieve increasingly stringent effluent permits. The use of photobioreactors (PBRs) and the separation of hydraulic retention and solids residence time (HRT/SRT) further enables increased biomass in a reduced physical footprint while allowing operational parameters (e.g., SRT) to select for desired functional communities. However, as algal technology transitions to full-scale, there is a need to understand the effect of operational and environmental parameters on complex microbial dynamics among mixotrophic microalgae, bacterial groups, and pests (i.e., grazers and pathogens) and to implement robust process controls for stable long-term performance. Here, we examine a full-scale, intensive WRRF utilizing mixed microalgae for tertiary treatment in the US (EcoRecover, Clearas Water Recovery Inc.) during a nine-month monitoring campaign. We investigated the temporal variations in microbial community structure (18S and 16S rRNA genes), which revealed that stable system performance of the EcoRecover system was marked by a low-diversity microalgal community (D INVSIMPSON = 2.01) dominated by Scenedesmus sp. (MRA = 55 %-80 %) that achieved strict nutrient removal (effluent TP < 0.04 mg·L -1 ) and steady biomass concentration (TSS monthly avg . = 400–700 mg·L −1 ). Operational variables including pH, alkalinity, and influent ammonium (NH 4 + ), correlated positively (p < 0.05, method = Spearman) with algal community during stable performance. Further, the use of these parameters as operational controls along with N/P loading and SRT allowed for system recovery following upset events. Importantly, the presence or absence of bacterial nitrification did not directly impact algal system performance and overall nutrient recovery, but partial nitrification (potentially resulting from NO 2 − accumulation) inhibited algal growth and should be considered during long-term operation. The microalgal communities were also adversely affected by zooplankton grazers (ciliates, rotifers) and fungal parasites (Aphelidium), particularly during periods of upset when algal cultures were experiencing culture turnover or stress conditions (e.g., nitrogen limitation, elevated temperature). Altogether, the active management of system operation in order to maintain healthy algal cultures and high biomass productivity can result in significant periods (>4 months) of stable system performance that achieve robust nutrient recovery, even in winter months in northern latitudes (WI, USA).

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2023

Modulation of xanthophyll cycle impacts biomass productivity in the marine microalga Nannochloropsis

Life on earth depends on photosynthetic primary producers that exploit sunlight to fix CO 2 into biomass. Approximately half of global primary production is associated with microalgae living in aquatic environments. Microalgae also represent a promising source of biomass to complement crop cultivation, and they could contribute to the development of a more sustainable bioeconomy. Photosynthetic organisms evolved multiple mechanisms involved in the regulation of photosynthesis to respond to highly variable environmental conditions. While essential to avoid photodamage, regulation of photosynthesis results in dissipation of absorbed light energy, generating a complex trade-off between protection from stress and light-use efficiency. This work investigates the impact of the xanthophyll cycle, the light-induced reversible conversion of violaxanthin into zeaxanthin, on the protection from excess light and on biomass productivity in the marine microalgae of the genus Nannochloropsis. Zeaxanthin is shown to have an essential role in protection from excess light, contributing to the induction of nonphotochemical quenching and scavenging of reactive oxygen species. On the contrary, the overexpression of zeaxanthin epoxidase enables a faster reconversion of zeaxanthin to violaxanthin that is shown to be advantageous for biomass productivity in dense cultures in photobioreactors. These results demonstrate that zeaxanthin accumulation is critical to respond to strong illumination, but it may lead to unnecessary energy losses in light-limiting conditions and accelerating its reconversion to violaxanthin provides an advantage for biomass productivity in microalgae.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Moderate high temperature is beneficial or detrimental depending on carbon availability in the green alga Chlamydomonas reinhardtii

High temperatures impair plant growth and reduce agricultural yields, but the underlying mechanisms remain elusive. The unicellular green alga Chlamydomonas reinhardtii is an excellent model to study heat responses in photosynthetic cells due to its fast growth rate, many similarities in cellular processes to land plants, simple and sequenced genome, and ample genetic and genomics resources. Chlamydomonas grows in light by photosynthesis and with externally supplied acetate as an organic carbon source. Understanding how organic carbon sources affect heat responses is important for the algal industry but remains understudied. We cultivated wild-type Chlamydomonas under highly controlled conditions in photobioreactors at 25 °C (control), 35 °C (moderate high temperature), or 40 °C (acute high temperature) with or without constant acetate supply for 1 or 4 day. Treatment at 35 °C increased algal growth with constant acetate supply but reduced algal growth without sufficient acetate. The overlooked and dynamic effects of 35 °C could be explained by induced acetate uptake and metabolism. Heat treatment at 40 °C for more than 2 day was lethal to algal cultures with or without constant acetate supply. Furthermore, our findings provide insights to understand algal heat responses and help improve thermotolerance in photosynthetic cells.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2026

Effects of Media Nutrient Variation on Microalgae Productivity and Economics During Semi-Continuous Cultivation

The development of large-scale microalgae growth for biofuel production is currently limited by the cost of biomass production. However, new approaches to infrastructure and cultivation practices are bringing the field closer to realization. Macronutrients in the cultivation media contribute significant costs, especially since their concentrations have not been optimized for specific strains and conditions. Environmental photobioreactors (ePBRs) were used to simulate cultivation under outdoor conditions, during which the nitrogen and phosphorus levels in the media were varied. The growth of two potential biofuel production strains, Picochlorum celeri and Tetraselmis striata, with varying nutrient inputs during summer and winter scripts, respectively, was studied. This study demonstrated that nitrogen and phosphorus in f/2 media could be reduced by more than 60% from the standard formulation, while maintaining growth rates in a semi-continuous harvesting approach. Experiments comparing the standard and reduced nutrient input concentrations were also conducted for both species in 820 L outdoor raceway ponds, in Mesa, AZ. P. celeri grown in these ponds in October had a growth rate of 10.6 ± 0.7 g/m 2 /day and 10.6 ± 0.3 g/m 2 /day for the standard and low-nutrient P. celeri ponds, respectively. T. striata grown in April–May had a growth rate of 16.6 ± 1.4 g/m 2 /day for the standard nutrient input ponds and 17.4 ± 1.1 g/m 2 /day for the low-nutrient input ponds, and in October 14.5 ± 0.6 g/m 2 /day for standard nutrient ponds and 14.4 ± 0.6 g/m 2 /day for low-nutrient ponds. These outdoor data therefore confirmed the indoor ePBR data. Techno-economic analysis shows that, if high growth rates can be attained at lower nutrient concentrations, a reduction of at least 60% in nutrient costs can be achieved. Such results highlight the importance of managing macronutrient media inputs, as these have a considerable contribution to biomass production costs in large-scale facilities. The analysis also points to the importance of maintaining high spent medium recycling rates in an industrial deployment, so as to minimize the losses of nitrogen and phosphorus compounds.

09 BIOMASS FUELS↗
DOE OSTI2023

Towards an Integrated Process for CO 2 Capture and Utilization: Cultivation of Scenedesmus acutus Using Gaseous CO 2 and NH 3

Integrating CO 2 scrubbing from flue gas with its utilization in algae cultivation represents a potential means of lowering the cost of CO 2 capture. Towards this goal, this study sought to assess the feasibility of using gaseous a CO 2 /NH 3 stream, derived from CO 2 capture using aqueous ammonia, as a C- and N-source for algae cultivation. Scenedesmus acutus was cultured in 800 mL photobioreactors using gaseous CO 2 /NH 3 in mole ratios varying from 7 to 18. Excellent growth of Scenedesmus acutus was observed, the average growth rate for CO 2 /NH 3 = 10 of 0.171 ± 0.015 g/L·day exceeding that obtained using 1% CO 2 /N 2 and urea as the N-source (0.099 ± 0.28 g/L·day). Under optimal growth conditions (CO 2 /NH 3 mole ratio of 10), CO 2 utilization ranged from 57% to 72%, while the NH 3 utilization was >90%. The CO 2 /NH 3 feed rate was also found to exert a significant effect on algae productivity, excessive feed rates leading to accumulation of NH 3 in the culture at concentrations that were toxic to the algae. Consequently, to avoid the toxic effects of high NH 3 concentrations (>2.0 mM), it proved necessary to balance the NH 3 supply with the algae growth rate so that excessive NH 3 accumulation was prevented. This indicates that for practical applications, a CO 2 /NH 3 feed control strategy would be required that takes into account the ammonium ion concentration in solution and the pH so as to avoid significant concentrations of free NH 3 . Furthermore, analysis of the harvested biomass revealed a high protein (≥ 47 wt%) and a low ash content (< 3.6 wt%), suggesting it would be well suited for use as animal feed or as a feedstock for the production of bioplastics.

03 NATURAL GAS↗
DOE OSTI2021

Pigment modulation in response to irradiance intensity in the fast-growing alga Picochlorum celeri

Picochlorum celeri has among the fastest photoautotrophic growth rates (~2 h doubling time in optimal conditions) reported to date for a marine alga. This study comprehensively analyzes the levels of Picochlorum celeri photosynthetic pigments, which can reach up to ~13% of the total particulate organic carbon (POC) under light-limiting conditions. The main Picochlorum celeri pigments identified include: chlorophyll a, chlorophyll b, lutein, β-carotene, canthaxanthin, violaxanthin, neoxanthin, zeaxanthin and antheraxanthin. The ketocarotenoid canthaxanthin can accumulate up to 120 mg L -1 in Picochlorum celeri liquid culture; ~12% of it was found in an extracellular polysaccharide matrix. Using a solar-simulating automated photobioreactor, we monitor the photoacclimation of cultures maintained in unshaded conditions (<0.5 μg mL -1 of total chlorophyll) through transitions from high irradiance (1000 μmoles photosynthetically active radiation (PAR) m -2 s -1 ) to low irradiance (60 μmoles PAR m -2 s -1 ), and conversely from low to high irradiance. Canthaxanthin and zeaxanthin accumulation are among the most rapid modulation responses when cultures are shifted from low to high irradiance. The violaxanthin, antherazanthin, and zeaxanthin (VAZ) pool is ~3-fold higher in high-light cultures, suggesting that the VAZ cycle combined with a dramatic reduction in chlorophyll levels are among the major mechanisms used in Picochlorum celeri to efficiently acclimate to high-irradiance levels. Responsive pigment modulation was also observed in denser cultures (~0.65 g L –1 ) grown under a diel cycle (pond-mimicking conditions). Furthermore, this research provides unique insights into the dynamics of pigment modulation and photoacclimation in response to changing irradiance in a biotechnologically promising alga and will inform future pigment engineering strategies to further improve light capture and biomass accumulation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Lead tolerance and bioremoval by four strains of green algae from Nigerian fish ponds

Heavy metals from industry pose threats to human and environmental health. Microalgae are prime candidates to remove heavy metals from water and soil due to their metal-binding affinities, high growth rates, and contamination tolerance. Strain selection and composition of resulting contaminated biomass must be considered for downstream processing. In this study, we assessed the ability of four microalgae strains (Chlorophyceae) that were isolated from contaminated Nigerian fish ponds to take up dissolved lead at environmentally relevant concentrations (<0.05 mg L -1 – WHO standard). In a microplate assay, all strains grew in the presence of lead, tolerating concentrations up to 60 ppm, though the addition of dissolved lead affected the growth of the four strains. In a photobioreactor assay, all strains demonstrated the ability to take up dissolved lead, with removal efficiencies ranging from 86 to 93%. The concentrations of lead in contaminated biomass, removal efficiencies, and bioconcentration factors were greatest in one of the isolates Chlorella sp. MOW 12. Our results reinforce the fact that live microalgae have a greater biosorption capacity. Lead recovery from freeze-dried biomass using five different reagents showed that a 0.1 M hydrochloric acid solution was the most effective, with 83.4% recovery. We also showed that presence of lead in the culture medium resulted in an increase in the quantity of protein in the biomass. Elemental analysis of lead-bound biomass showed an increase in elements including boron and copper and a decrease in cobalt and sodium. Synergystic and antagonystic effect in uptake of other elements was strain specific, suggesting the possibility of tailored wastewater treatments depending on specific requirements.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Development of integrated screening, cultivar optimization, and verification research (DISCOVR): A coordinated research-driven approach to improve microalgal productivity, composition, and culture stability for commercially viable biofuels production

To address major knowledge gaps and barriers to the commercial development of algal biomass for biofuels and co-products, a collaborative consortium, Development of Integrated Screening, Cultivar Optimization, and Verification Research (DISCOVR), was established in 2016. Funded by the U.S. Department of Energy (DOE) Bioenergy Technologies Office (BETO), this consortium constitutes a partnership between four DOE national laboratories - Pacific Northwest National Laboratory (PNNL), Los Alamos National Laboratory (LANL), the National Renewable Energy Laboratory (NREL), and Sandia National Laboratories (SNL) - and the Arizona Center for Algae Technology and Innovation (AzCATI) at Arizona State University. To address the barriers of strain selection for achieving high seasonal productivities with a suitable composition and culture resilience, a tiered strain down-selection pipeline is implemented. At Tier I, the temperature and salinity tolerance of strains is determined in flask cultures; at Tier II, the areal biomass productivity and composition is determined in climate-simulation photobioreactors; at Tier III, the productivity and culture stability are determined in outdoor raceways. The top performing strains move forward to long-term testing at the algae testbed site at AzCATI to generate annual biomass productivity data. Concurrent to the strain down-selection in the DISCOVR pipeline, hypotheses for increasing biomass productivity, shifting biomass composition to enhance intrinsic value, and improving culture stability and resistance to pests are also tested. Techno-economic analyses are carried out to determine whether promising findings from laboratory studies or proposed modifications in outdoor pond cultivation conditions translate into reductions in the minimum biomass selling price (MBSP). Notably, in the three years following the launch of DISCOVR, annual biomass productivity has increased from 11.7 to 17.6 g m -2 day -1 , resulting in an MBSP decrease from 824 to 611 $ ton -1 .

09 BIOMASS FUELS↗
DOE OSTI2023

Outdoor annual algae productivity improvements at the pre-pilot scale through crop rotation and pond operational management strategies

The Development of Integrated Screening, Cultivar Optimization, and Verification Research (DISCOVR) collaborative consortium operated pre-pilot scale outdoor ponds to deliver much-needed multi-year, long-term and consistent, algae cultivation data relevant to understanding the current state of technology in terms of expected seasonal algae biomass productivity. Over the course of four years from 2018 to 2021, twelve identical 4.2 m 2 mini-ponds were run in triplicate sets to test strains and operational strategies demonstrated in small-, indoor photobioreactors, in pursuit of increasing overall algae areal productivity and projected farm yield. Fourteen different cultivars derived from a strain screening pipeline were tested. Through deliberate seasonal crop rotation and improvements in operational strategies, annual biomass productivity increased from 11.6 to 17.6 g m -2 day -1 , a > 50% increase over the 2018 baseline. Both brackish and marine strains were included and four out of the fourteen strains consistently yielded high productivity across multiple years; brackish strains Monoraphidium minutum (26BAM) and Scenedesmus obliquus (UTEX393), and marine strains Tetraselmis striata (LANL1001) and Picochlorum celeri (TG2). These freely available datasets, which represent nearly complete annual daily coverage of cultivation metrics including weather, pond temperature and pH, nutrients, and productivity, are unique in the public domain and seek to fill agronomic and operational knowledge gaps to help in the eventual commercialization of algal biofuels and bioproducts.

09 BIOMASS FUELS↗
DOE OSTI2023

DISCOVR strain pipeline screening – Part I: Maximum specific growth rate as a function of temperature and salinity for 38 candidate microalgae for biofuels production

Here, to identify high productivity strains for microalgal biofuels generation, the maximum specific growth rate of 38 strains was measured as a function of salinity (i.e., 5, 15, and 35 PSU) and temperature (i.e., at 8 temperatures along a linear gradient from ca. 5 to 45°C) to determine the most suitable growth medium salinity and best growing season, respectively, for outdoor raceway pond cultivation. The following strains were evaluated: Agmenellum quadruplicatum UTEX 2268, Anabaena sp. ATCC 33081, Arthrospira fusiformis UTEX 2721, Arthrospira platensis UTEX 3086, Chlorella vulgaris NREL 4-C12, Chlorella autotrophica CCMP 243, Chlorella sorokiniana DOE1044, Chlorella sorokiniana DOE 1116, Chlorella sorokiniana DOE 1412 (UTEXB3016), Chlorella vulgaris LRB AZ-1201, Chlorococcum littorale UTEX 117, Chlorococcum sp. UTEX-B P7, Chloromonas reticulata CCALA 870, Coelastrella sp. DOE 0202, Cyanobacterium sp. AB1, Micractinium reisseri NREL 14-F2, Microchloropsis gaditana CCMP1894, Microchloropsis salina CCMP 1776, Monoraphidium sp. MONOR1, Monoraphidium minutum 26B-AM, Nannochloropsis oceanica CCAP 849/10, Oscillatoria cf. priestleyi CCMEE 5020.1-1, Picochlorum celeri TG2-WT-CSM/EMRE, Picochlorum oklahomensis CCMP 2329, Picochlorum renovo NREL 39-A8, Picochlorum soloecismus DOE 101, Porphyridium cruentum CCMP 675, Scenedesmus acutus LRB-AP-0401, Scenedesmus obliquus DOE 0152.z, Scenedesmus obliquus UTEX393,Scenedesmus rubescens NREL 46B-D3, Scenedesmus sp. IITRIND2, Stichococcus minor CCMP 819, Stichococcus minutus CCALA 727, Synechococcus elongatus UTEX2973.1, Tetraselmis striata LANL 1001, Tisochrysis lutea CCMP 1324, and Tribonema minus UTEXB3156. For each strain, the identity and the presence of bacterial cohorts was determined using 18S and 16S rDNA sequencing, respectively. The maximum specific growth rate versus temperature data were also used to determine the activation energies (Arrhenius equation) for most strains. For all strains, the measured salinity and temperature tolerance data were compared to those reported in the literature. The fastest growing strains were down-selected for subsequent biomass productivity measurements in climate-simulation photobioreactors, as reported in the next paper in the issue.

18S and 16S rDNA sequencing↗
DOE OSTI2022

Mineral-hydrogel composites for mitigating harmful algal bloom and supplying phosphorous for photo-biorefineries

Harmful algal blooms (HAB) are a major environmental concern in eutrophic aquatic systems. To mitigate HABs and recover the phosphorus that drives algal growth, this study developed hydrogel composites seeded with calcium phosphate and wollastonite particles, which first adsorb phosphate (P) and then precipitate it as calcium phosphate. Using a fast-growing cyanobacterium, Synechococcus elongatus 2973, as a model microalga, we found that the mineral-hydrogel composites reduced dissolved P in BG11 media from 5.1 mg/L to 0.31 mg/L, initially reducing the biomass growth rate by up to 73 % and ultimately reducing the total biomass concentration by 75 %. When applied to municipal wastewater and agricultural run-off, the composites removed 96 % and 91 % of the dissolved P, respectively. Moreover, when the recovered P-enriched composites were reused as a slow-release bio-compatible fertilizer in a photobioreactor, they effectively supported algal growth without blocking light and interfering with photosynthesis. The P-enriched composites could tune the P concentration in the culture medium and significantly promote algal lipid accumulation. Finally, this study demonstrates the mineral-hydrogel composites' potential to treat point sources of P pollution and subsequently facilitate photoautotrophic biofuel production as a nutrient, effectively recycling the captured P.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Machine learning-informed and synthetic biology-enabled semi-continuous algal cultivation to unleash renewable fuel productivity

Abstract Algal biofuel is regarded as one of the ultimate solutions for renewable energy, but its commercialization is hindered by growth limitations caused by mutual shading and high harvest costs. We overcome these challenges by advancing machine learning to inform the design of a semi-continuous algal cultivation (SAC) to sustain optimal cell growth and minimize mutual shading. An aggregation-based sedimentation (ABS) strategy is then designed to achieve low-cost biomass harvesting and economical SAC. The ABS is achieved by engineering a fast-growing strain, Synechococcus elongatus UTEX 2973, to produce limonene, which increases cyanobacterial cell surface hydrophobicity and enables efficient cell aggregation and sedimentation. SAC unleashes cyanobacterial growth potential with 0.1 g/L/hour biomass productivity and 0.2 mg/L/hour limonene productivity over a sustained period in photobioreactors. Scaling-up the SAC with an outdoor pond system achieves a biomass yield of 43.3 g/m 2 /day, bringing the minimum biomass selling price down to approximately $281 per ton.

09 BIOMASS FUELS↗
DOE OSTI2022

Photobiological production of high-value pigments via compartmentalized co-cultures using Ca-alginate hydrogels

Abstract Engineered cyanobacterium Synechococcus elongatus can use light and CO 2 to produce sucrose, making it a promising candidate for use in co-cultures with heterotrophic workhorses. However, this process is challenged by the mutual stresses generated from the multispecies microbial culture. Here we demonstrate an ecosystem where S. elongatus is freely grown in a photo-bioreactor (PBR) containing an engineered heterotrophic workhorse (either β-carotene-producing Yarrowia lipolytica or indigoidine-producing Pseudomonas putida ) encapsulated in calcium-alginate hydrogel beads. The encapsulation prevents growth interference, allowing the cyanobacterial culture to produce high sucrose concentrations enabling the production of indigoidine and β-carotene in the heterotroph. Our experimental PBRs yielded an indigoidine titer of 7.5 g/L hydrogel and a β-carotene titer of 1.3 g/L hydrogel, amounts 15–22-fold higher than in a comparable co-culture without encapsulation. Moreover, 13 C-metabolite analysis and protein overexpression tests indicated that the hydrogel beads provided a favorable microenvironment where the cell metabolism inside the hydrogel was comparable to that in a free culture. Finally, the heterotroph-containing hydrogels were easily harvested and dissolved by EDTA for product recovery, while the cyanobacterial culture itself could be reused for the next batch of immobilized heterotrophs. This co-cultivation and hydrogel encapsulation system is a successful demonstration of bioprocess optimization under photobioreactor conditions.

59 BASIC BIOLOGICAL SCIENCES↗