Thomas A. Dempster

Characterization of the growth and lipid content of the diatom Chaetoceros muelleri

Chaetoceros muelleri (Schütt) was cultured on a thermal gradient plate, subjected to two media types with a range of specific conductances, and evaluated for growth and neutral lipid accumulation. Growth was measured directly by daily changes in cell numbers and indirectly by changes in optical density at 750 nm. C. muelleri exhibited a growth rate of at least two doublings day-1 over broad temperature (20 to 35 °C) and conductance ranges (10 to over 60 mS cm-1) and the optimum growth rate approached 4.0 doublings day-1 at 30 °C and a conductance of 25 mS cm-1. Intracellular neutral lipid storage was evaluated with fluorometry and epifluorescent microscopy using the fluorochrome Nile Red. Gravimetric analysis revealed a total lipid content in nitrogen-depleted cultures of C. muelleri of over 400 mg L-1, five to seven times that observed in nitrogen-replete cultures. Based on its high growth rate, tolerance to a broad range of temperatures and specific conductances, and large quantity of intracellular lipid, C. muelleri may have potential for exploitation as a renewable precursor to liquid fuels or as a lipid source.

Algal Biomass Constituent Analysis: Method Uncertainties and Investigation of the Underlying Measuring Chemistries

Algal biomass compositional analysis data form the basis of a large number of techno-economic process analysis models that are used to investigate and compare different processes in algal biofuels production. However, the analytical methods used to generate these data are far from standardized. This work investigated the applicability of common methods for rapid chemical analysis of biomass samples with respect to accuracy and precision. This study measured lipids, protein, carbohydrates, ash, and moisture of a single algal biomass sample at 3 institutions by 8 independent researchers over 12 separate workdays. Results show statistically significant differences in the results from a given analytical method among laboratories but not between analysts at individual laboratories, suggesting consistent training is a critical issue for empirical analytical methods. Significantly different results from multiple lipid and protein measurements were found to be due to different measurement chemistries. We identified a set of compositional analysis procedures that are in best agreement with data obtained by more advanced analytical procedures. The methods described here and used for the round robin experiment do not require specialized instrumentation, and with detailed analytical documentation, the differences between laboratories can be markedly reduced.

EFFECTS OF ENVIRONMENTAL CONDITIONS ON GROWTH AND LIPID ACCUMULATION IN NITZSCHIA COMMUNIS (BACILLARIOPHYCEAE)

Microalgae from natural saline habitats in the southwestern United States were isolated and screened for rapid growth potential and ability to store intracellular neutral lipid, a precursor for renewable liquid fuel sources. After preliminary screening of isolated strains, Nitzschia communis (Rabenhorst) was selected for optimization of growth rate and lipid yield. Nitzschia communis was subjected to two media types (SERI I, II) with different major ion compositions designed to mimic natural saline groundwater aquifers in the arid Southwest. Growth rates were determined through 4 days of logarithmic growth, followed by 2 days of physiological stress (nitrogen deprivation) and subsequent measurement of neutral lipid accumulation. Poststress intracellular neutral lipid increases were documented by utilizing the fluorochrome Nile Red with fluorometric analysis and epifluorescent microscopy. Growth rate was slightly higher in SERI Type I medium, whereas lipid yield was higher in SERI Type II medium. Rapid growth and appreciable lipid yields were observed over a broad range of temperatures (20°–30° C) and specific conductances (10–70 mS·cm−1). The highest lipid yields were observed at elevated conductances (between 40 mS·cm−1 and 70 mS·cm−1). Substitution of individual salt concentrations from SERI Type II into SERI Type I medium indicated that increased lipid yield in Type I medium was associated with a reduction in MgCl2 concentration.

Strain, biochemistry, and cultivation-dependent measurement variability of algal biomass composition.

Accurate compositional analysis in biofuel feedstocks is imperative; the yields of individual components can define the economics of an entire process. In the nascent industry of algal biofuels and bioproducts, analytical methods that have been deemed acceptable for decades are suddenly critical for commercialization. We tackled the question of how the strain and biochemical makeup of algal cells affect chemical measurements. We selected a set of six procedures (two each for lipids, protein, and carbohydrates): three rapid fingerprinting methods and three advanced chromatography-based methods. All methods were used to measure the composition of 100 samples from three strains: Scenedesmus sp., Chlorella sp., and Nannochloropsis sp. The data presented point not only to species-specific discrepancies but also to cell biochemistry-related discrepancies. There are cases where two respective methods agree but the differences are often significant with over- or underestimation of up to 90%, likely due to chemical interferences with the rapid spectrophotometric measurements. We provide background on the chemistry of interfering reactions for the fingerprinting methods and conclude that for accurate compositional analysis of algae and process and mass balance closure, emphasis should be placed on unambiguous characterization using methods where individual components are measured independently.

Rapid Characterization of Microalgae and Microalgae Mixtures Using Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)

Current molecular methods to characterize microalgae are time-intensive and expensive. Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) may represent a rapid and economical alternative approach. The objectives of this study were to determine whether MALDI-TOF MS can be used to: 1) differentiate microalgae at the species and strain levels and 2) characterize simple microalgal mixtures. A common protein extraction sample preparation method was used to facilitate rapid mass spectrometry-based analysis of 31 microalgae. Each yielded spectra containing between 6 and 56 peaks in the m/z 2,000 to 20,000 range. The taxonomic resolution of this approach appeared higher than that of 18S rDNA sequence analysis. For example, two strains of Scenedesmus acutus differed only by two 18S rDNA nucleotides, but yielded distinct MALDI-TOF mass spectra. Mixtures of two and three microalgae yielded relatively complex spectra that contained peaks associated with members of each mixture. Interestingly, though, mixture-specific peaks were observed at m/z 11,048 and 11,230. Our results suggest that MALDI-TOF MS affords rapid characterization of individual microalgae and simple microalgal mixtures.

Analysis and modeling of Nannochloropsis growth in lab, greenhouse, and raceway experiments

Efficient production of algal biofuels could reduce dependence on foreign oil by providing a domestic renewable energy source. Moreover, algae-based biofuels are attractive for their large oil yield potential despite decreased land use and natural resource (e.g., water and nutrients) requirements compared to terrestrial energy crops. Important factors controlling algal lipid productivity include temperature, nutrient availability, salinity, pH, and the light-to-biomass conversion rate. Computational approaches allow for inexpensive predictions of algae growth kinetics for various bioreactor sizes and geometries without the need for multiple, expensive measurement systems. Parametric studies of algal species include serial experiments that use off-line monitoring of growth and lipid levels. Such approaches are time consuming and usually incomplete, and studies on the effect of the interaction between various parameters on algal growth are currently lacking. However, these are the necessary precursors for computational models, which currently lack the data necessary to accurately simulate and predict algae growth. In this work, we conduct a lab-scale parametric study of the marine alga Nannochloropsis salina and apply the findings to our physics-based computational algae growth model. We then compare results from the model with experiments conducted in a greenhouse tank and an outdoor, open-channel raceway pond. Results show that the computational model effectively predicts algae growth in systems across varying scale and identifies the causes for reductions in algal productivities. Applying the model facilitates optimization of pond designs and improvements in strain selection.

Influence of biochemical composition during hydrothermal liquefaction of algae on product yields and fuel properties

Hydrothermal liquefaction (HTL) of nine algae species were performed at two reaction temperatures (280 and 320°C) to compare the effect of their biomass composition on product yields and properties. Results obtained after HTL indicate large variations in terms of bio-oil yields and its properties. The maximum bio-oil yield (66wt%) was obtained at 320°C with a high lipid containing algae Nannochloropsis. The higher heating value of bio-oils ranged from 31 to 36MJ/kg and around 50% of the bio-oils was in the vacuum gas oil range while high lipid containing algae Nannochloropsis contained a significant portion (33-42%) in the diesel range. A predictive relationship between bio-oil yields and biochemical compositions was developed and showed a broad agreement between predictive and experimental yields. The aqueous phases obtained had high amount of TOC (12-43g/L), COD (35-160g/L), TN (1-18g/L), ammonium (0.34-12g/L) and phosphate (0.7-12g/L).

Catalytic upgrading of bio-oil produced from hydrothermal liquefaction of Nannochloropsis sp.

Upgrading of bio-oil obtained from hydrothermal liquefaction (HTL) of algae is necessary for it to be used as a fuel. In this study, bio-oil obtained from HTL of Nannochloropsis sp. was upgraded using five different catalysts (Ni/C, ZSM-5, Ni/ZSM-5, Ru/C and Pt/C) at 300 °C and 350 °C. The upgraded bio-oil yields were higher at 300 °C; however, higher quality upgraded bio-oils were obtained at 350 °C. Ni/C gave the maximum upgraded bio-oil yield (61 wt%) at 350 °C. However, noble metal catalysts (Ru/C and Pt/C) gave the better upgraded bio-oils in terms of acidity, heating values, and nitrogen values. The higher heating value of the upgraded bio-oils ranged from 40 to 44 MJ/kg, and the nitrogen content decreased from 5.37 to 1.29 wt%. Most of the upgraded bio-oils (35-40 wt%) were in the diesel range. The major components present in the gaseous products were CH4, CO, CO2 and lower alkanes.

From benchtop to raceway : spectroscopic signatures of dynamic biological processes in algal communities.

The search is on for new renewable energy and algal-derived biofuel is a critical piece in the multi-faceted renewable energy puzzle. It has 30x more oil than any terrestrial oilseed crop, ideal composition for biodiesel, no competition with food crops, can be grown in waste water, and is cleaner than petroleum based fuels. This project discusses these three goals: (1) Conduct fundamental research into the effects that dynamic biotic and abiotic stressors have on algal growth and lipid production - Genomics/Transcriptomics, Bioanalytical spectroscopy/Chemical imaging; (2) Discover spectral signatures for algal health at the benchtop and greenhouse scale - Remote sensing, Bioanalytical spectroscopy; and (3) Develop computational model for algal growth and productivity at the raceway scale - Computational modeling.