1. Exposing C.reinhardtii to Anaerobic Atmospheric Conditions to Enhance Hydrogen Production Chrisitna George Period 0,1 Even June 1, 2009

2. Need http://www.physicalgeography.net/fundamentals/images/co2_atmosphere.jpg

3. Need http://www.tspusa.com/images/TICimages/alternativefuels1.gif

4. Knowledge Base Autotrophic organism that produces hydrogen in sulfur deprived anaerobic conditions Autotrophic organism that produces hydrogen in sulfur deprived anaerobic conditions http://www.sciencedaily.com/images/2009/03 /090324171556-large.jpg C.reinahrdtii

5. Knoweldge Base http://generalhorticulture.tamu.edu/lectsupl/Light/p54f1.gif Light intensities v. the CO2 exchange when measured by the photosynthetic rate

6. Knowledge Base Anaerobic Conditions: the absence of oxygen Anaerobic Conditions: the absence of oxygen http://www.ilmvac.co.uk/content/products/Anaerobic-container-p112555-01.html

7. Isoamylase Gene Important gene for starch accumulation which is important for hydrogen production (Posewitz, 2005). Important gene for starch accumulation which is important for hydrogen production (Posewitz, 2005). http://www.mrc-lmb.cam.ac.uk/genomes/date/1bf2.gif

8. Literature Review Rosenbaum, Miriam (2005) Rosenbaum, Miriam (2005) ‘Utilizing the Green Alga C.reinhardtii for Microbial Electricity Generation” Direct electricity generation from microbial photosynthetic activity Oxidative depletion of hydrogen, photosynthetically produced by C.reinahrdtii under sulfur deprived conditions, by polymer coated electro catalytic electrodes

9. Literature Review Melis, Anastasios (2000) Melis, Anastasios (2000) “Sustained Photobiological Hydrogen Gas Production upon Reversible Inactivation of oxygen Evolution in C.reinhardtii” Method circumvents the severe oxygen sensitivity of the reversible hydrogenase by temporarily separating photosynthetic oxygen evolution and carbon dioxide accumulation

10. Literature Review Posewitz, Matthew C. (2004) Posewitz, Matthew C. (2004) “Hydrogen Photoproduction is Attenuated by Disruption of an Isoamylase Gene in C.reinhardtii” STA7 is an isoamylase gene in C.reinahrdtii, which is important for starch accumulation and metabolism, which plays an important role in hydrogen photoproduction

11. Literature Review Logan, Bruce E. (2006) Logan, Bruce E. (2006) “Using Algae and other Biomass for Hydrogen Production in a Modified Microbial Fuel Cell” A small voltage was applied (.25 V) to the algae, which generated pure hydrogen gas at the cathode

12. Hypotheses Alternate: A short period of anaerobic conditions will increase the hydrogen production Alternate: A short period of anaerobic conditions will increase the hydrogen production Null: The duration and frequency of anaerobic conditions will have no effect on the amount of hydrogen produced Null: The duration and frequency of anaerobic conditions will have no effect on the amount of hydrogen produced

13. Purpose Therefore, the purpose of this experiment is to find the optimal duration and frequency of anaerobic exposure needed to optimize C.reinhardtii hydrogen production Therefore, the purpose of this experiment is to find the optimal duration and frequency of anaerobic exposure needed to optimize C.reinhardtii hydrogen production

14. Exposing C.reinhardtii to Anaerobic Atmospheric Conditions to Enhance Hydrogen Production Problem: What is the optimal duration and frequency of anaerobic exposure needed to optimize C.reinhardtii hydrogen production? Control Groups: The Algae growth medium, Temperature, and light intensity Independent Variable: Growth of C.reinhardtii and hydrogen produced Dependent Variable: Carbon dioxide levels, pH levels All data will statistically analyzed by SPSS, by an ANOVA followed by a Sheffe Post Hoc Test. Data will then be put in line graphs. -The hydrogen gas produced will be measured using an H2Scan hydrogen detector. -A fluorometer will be used to measure the photosynthetic rate. 6 hours of anaerobic atmospheric conditions 12 hours of anaerobic atmospheric conditions 24 hours of anaerobic atmospheric conditions -Carbon dioxide concentration measured using a Pasco GLX Xplorer -pH levels measured using pH paper -growth of C.reinhardtii measured using a Spectrophotometer

15. Do Ability C.reinhardtii: Can be grown in the lab as done in previous years C.reinhardtii: Can be grown in the lab as done in previous years Light Intensities: Different bulbs can be purchased and used in the school lights Light Intensities: Different bulbs can be purchased and used in the school lights Microbial Fuel Cell: A simple fuel cell is owned by the school Microbial Fuel Cell: A simple fuel cell is owned by the school

16. Budget Christina George VendorOrder #QuantityDescriptionPriceTotal price Carolina Biological1520404C.reinhardtii9.50 (ea)38 Carolina Biological1537522Medium16.85 (ea)33.7 Ilmavc112555-011anaerobic container Warehouse Lighting WS-110344 1light99.99 VibraFlobubbler fishsupply.com SPAEH- 21645 1sump-pump139.99 HomeDepottubing HomeDepot1wooding base Corning1stirrer A.W. Sperry1photometer fishsupply.com stm- heg119 6 1thermo sensor12.99 fishsupply.com slnt-sdt02 1timer19.99 Aturner DesignsFluorometer h2Scan1HY-ALERTA™ 2600

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