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Megan Snyder and Sam Huynh

Studying the effect of different irradiances of light on the triacylglycerol yield in the marine diatom Thalassiosira pseudonana. Megan Snyder and Sam Huynh . Background Information. T. pseudonana is classified as phytoplankton (photosynthesizing marine organisms)

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Megan Snyder and Sam Huynh

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  1. Studying the effect of different irradiances of light on the triacylglycerol yield in the marine diatom Thalassiosira pseudonana Megan Snyder and Sam Huynh

  2. Background Information • T. pseudonana is classified as phytoplankton (photosynthesizing marine organisms) • Diatom (microscopic, unicellular, photosynthetic algae with a silica shell called a frustule) • Bacillariophyceae class (largest group of biomass producers)100,000+ species • High oil content (21-31%) by dry weight • Entire genome is sequenced • Oil helps it float on the water to photosynthesize http://www.diark.org/img/species_pict/Thalassiosira_pseudonana_CCMP1335

  3. Oil from diatom is a potential alternative fuel • Oil composed of lipids • Lipid: biological compound insoluble in water but soluble in organic solvents • Triacylglycerol (specific lipid) acts primarily as energy storage in diatoms (three fatty acids attached to one glycerol) • Triaclyglcerol is a non-polar, or neutral lipid • Polar lipids have a hydrophilic and a hydrophobic end; neutrals do not. • Polar lipids located in cell walls

  4. TriacylglycerolTransesterification • process to refine triacylglycerol; how does it become biodiesel? • Oil is purified and reacted with methanol or ethanol in the presence of a catalyst (such as KOH potassium hydroxide or NaOH sodium hydroxide); the triacylglycerol is transformed to form esters and glycerol – remaining esters are biodiesel

  5. Triacylglycerol Chemical Formula http://www.oliveoilsource.com/page/chemical-characteristics 3 Fatty Acids Glycerol

  6. Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) is a dye that collects in the neutral lipids of living cells • Fluoresces under a certain wavelength, then emits another wavelength • The intensity of the fluorescence is directly correlated to lipid content

  7. Purpose: increase the triacylglycerol content in the diatom using different light irradiances • Importance: increased triacylglycerol yields per diatom can make the process of making bio-fuel from algae more efficient and economically feasible

  8. Irradiance: measure of light that reaches the product surface • Why measure in irradiance and not intensity? Photosynthesis depends on the amount of light reaching the surface of the plant, not the intensity of the source

  9. Procedure • Diatoms will grow a plastic container under different irradiances of fluorescent light (test one irradiance at a time) • Sides of the container will be covered with aluminum foil to block out excess light • In previous experiments, irradiances of light have affected photosynthetic processes in T. pseudonana • Light on a 16 hour-on, 8 hour-off cycle, maintained by a light timer

  10. Grown in f/2 agar, a solution that provides nutrients for marine algae • pH measured periodically with a Vernier probe to make sure CO2 levels stay constant • Container inside Styrofoam box with ice packs to keep temperature constant at about 53°F • Nile red stain will be applied to a few sample diatoms after six days

  11. Data Collection • Use UV microscope equipped with a camera • Takes pictures of the cells fluorescing • Use Image J program to analyze level of fluorescence Inverted UV microscope Green Algae stained with Nile red- Triacylglycerol fluoresces yellow-gold, phospholipids fluoresce red http://www.chemistrytimes.com/Images/green_alga_with_nile_red.jpg http://accu-scope.net/3032_EPI_318CU.gif

  12. Bibliography • Chemical Education Digital Library (2007 a). Polar Lipids. Retrieved from: http://chemed.chem.wisc.edu/chempaths/GenChem-Textbook/Polar-Lipids-1019.html • Chemical Education Digital Library (2007 b). Nonpolar Lipids. Retrieved from: http://chemed.chem.wisc.edu/chempaths/GenChem-Textbook/Nonpolar-Lipids-1018.html • Chen, W., Zhang, C., Song, L., Sommerfield, M., & Hu, Q. (2009). A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae. Journal of Microbiological Methods, 77, 41-47. doi:10.1016/j.mimet.2009.01.001 • Demirbas, A. & Demirbas, F. M. (2010). Importance of algae oil as a source of biodiesel. Energy Conversion and Management. doi:10.1016/j.enconman.2010.06.055 • Epinoza, E. V., Nunez, R.M., Trees, C. C., et al (2007). Growth and accessory pigments to chlorophyll A ratios of Thalassiosira pseudonana (bacillariophyceae) cultured under different irradiances. Hidrobiologica, 17, 249-255. • Fromm, J. R. (1997). Introduction to Lipids. Retrieved from: http://www.3rd1000.com/chem301/chem302p.htm • Newman, S. (2008). Algae Biodiesel Engineering: Extracting Oil from Algae. Retrieved from: http://science.howstuffworks.com/environmental/green-science/algae-biodiesel2.htm • Reusch, W. (2004). Lipids. Retrieved from: http://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/lipids.htm#lipid1 • Zhukova, N. V. (2003). Changes in the lipid composition of Thalassiosira pseudonana during its life cycle. Russian Journal of Plant Physiology, 51(5), 702-707.

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