1. The Effect of Yeast and Temperature on the Alcohol Content of Zea Maize after Fermentation Madeline LaBelle—Warren Mott High School Keara Miley—Lake Shore High School

2. 2 Background Petroleum oil supplies are being depleted They are being used up rapidly A sufficient alternative is needed Biofuel can be used instead – Made from renewable biomass resources – Safe and environment friendly MMSTC

3. 3 Background Biofuel emits 51% less greenhouse gases than fossil fuel – More efficient methods of production Due to new technologies MMSTC

4. 4 Background Biofuel is made from the fermentation of plant matter Fermentation is the process in which yeast and enzymes break down sugars and convert them into alcohol MMSTC

5. 5 Background Alcohol content is directly related to how well a material can be used as a biofuel – The higher the better The higher the alcohol content, the hotter is burns and the better it is for a car’s engine MMSTC

6. 6 Background Cars have combustion engines How a combustion engine works – Increase in pressure from combustion expands the cylinder – Pushes piston and causes crankshaft to turn The hotter a material burns, the more combustion – More efficient MMSTC

7. 7 Background Different types of biofuel – Corn ethanol – Cane ethanol – Cellulosic ethanol Cane ethanol is most efficient Corn ethanol is the most common in America – Corn is the most abundant source here – No import taxes MMSTC

8. 8 Background Two different corn ethanol production processes – Dry milling – Wet milling Dry milling is the most common Wet milling is more extensive but produces more byproducts MMSTC

9. 9 Dry Milling Kernels ground up, enzymes and water added – Called mash – Enzymes break down starches into sugar Mash heated to reduce bacteria Mash is cooled and yeast added – Fermentation starts When fermentation finished, it is distilled – Distillation separates substances MMSTC

10. 10 Wet Milling Kernels soaked and the middle is removed – Used to produce corn oil Starch and gluten separated – Gluten used for animal feed Water and enzymes added to ferment – Remaining starch used to make corn syrup Mixture distilled and sent to oil companies MMSTC

11. 11 Terminology Zea maize – Corn Saccharomyces cerevisiae – Type of yeast Hydrometer – Tool used to measure alcohol content of a material DOE – Design of experiment MMSTC

12. 12 Purpose To find a way to make corn ethanol more efficient by increasing the alcohol content – Different temperatures during fermentation – Different amounts of yeast added to the mixture MMSTC

13. 13 Problem Statement What amount of Saccharomyces cerevisiae and what temperature during fermentation would yield the highest alcohol concentration in Zea maize? MMSTC

14. 14 Hypothesis Out of all amounts of Saccharomyces cerevisiae (1, 2, and 3 grams) and temperatures (22, 25, and 28 °C), 3 grams of yeast at a temperature of 22 °C will yield the highest concentration of alcohol in fermented corn. MMSTC

15. 15 Data Measured Dependent variable – The alcohol content of the fermented corn (%) Independent variables – The amount of yeast (grams) – Temperature during fermentation (°C) MMSTC

16. 16 Materials Active dry yeast (Saccharomyces cerevisiae) 21 half-bushels of corn 21 emptied two liter bottles Scale Funnel Balloons Blender Hydrometer Mesh colander Incubator (set at 22, 25, and 28 °C) MMSTC

17. 17 Blender Mesh Colander 2 Liter Bottle Balloon Scale Active Dry Yeast Corn Funnel Graduated Cylinder Hydrometer MMSTC

18. 18 Procedure Blend corn mixture together and measure alcohol content with hydrometer Add yeast to mixture (amount dependent on trial) Pour mixture into a 2 Liter bottle with the funnel MMSTC

19. 19 Place bottle in incubator (temperature dependent on trial) After four days, remove the corn mixture and measure alcohol content with hydrometer Subtract second alcohol measurement from the first MMSTC Procedure

20. 20 Record the value in the table Repeat for each combination of temperature and amount of yeast MMSTC Procedure

21. 21 Procedure Video MMSTC

22. 22 Data High and Low Values Temperature (°C)Amount of Yeast (g) -Standard+- + 222528123 MMSTC

23. 23 Data Design of Experiment Data OrderRunResultOrderRunResultOrderRunResult 1Standard61 61 6 2+ 72 72 8 3- 53 53 6 4Standard64 54 8 5+ -85 75 8 6- +56 56 6 7Standard77 67 6 MMSTC

24. 24 Data Standard Trials 667656666 MMSTC

25. 25 Data Runs First DOESecond DOEThird DOEAverage Temperature (°C) Amount of Yeast (g) ++7787.33 -+5565.33 +-8787.67 --5565.33 Grand Average: 6.145 MMSTC

26. 26 Observations DateObservation Oct 229 standard trials are set up. Corn and yeast blends are consistent. Oct 26 Corn and yeast blend produced CO 2 which can be seen by the balloon inflating. Oct 29All (+,-) trials produced CO 2 and the balloons inflated. Nov 2 Only 3 (+,+) trials produced CO 2. The balloon on the Trial 2 bottle did not inflate resulting in a redo of this trial. Nov 6All (-,-) trials produced CO 2 and the balloons inflated. Nov 10All (-,+) trials produced CO 2 and the balloons inflated. Nov 13Trial 17 (-,+) blend seemed more separated than the rest, but still produced CO 2 and stayed consistent with rest of trials. MMSTC

27. 27 Data Analysis and Interpretation A Two Factor DOE was used – Temperature (°C) – Amount of yeast (g) MMSTC

28. 28 What is a DOE? DOE stands for a Design of Experiment DOE measures the individual effects and combined effects of different factors Appropriate for this experiment because two different factors were manipulated at two levels MMSTC

29. 29 Data Analysis and Interpretation The effect of temperature on the alcohol content of fermented corn was 2.17 units As temperature increased, the alcohol content increased by 2.17 units MMSTC

30. 30 Data Analysis and Interpretation The effect of the amount of yeast on the alcohol content of fermented corn was -0.17 units As the amount of yeast increased, the alcohol content decreased by 0.17 units MMSTC

31. 31 Data Analysis and Interpretation The effect of temperature and yeast has an effect of -0.17 units There may be an interaction between the two effects because the slopes are not parallel MMSTC

32. 32 Determining Significance Any effect that is greater than two times the range of standards is deemed significant None of the variables were considered significant in this experiment MMSTC

33. 33 Conclusion The hypothesis was rejected Neither temperature during fermentation or amount of yeast had a significant effect on the alcohol content of corn after fermentation These results of this experiment agree with current work in this field MMSTC

34. 34 Why Did These Results Occur? The amount of yeast had no effect because yeast can only convert the sugars that are present in the mixture and after these sugars are converted can do no more Temperature will only speed up the process MMSTC

35. 35 What Will Have an Effect? The only factor that will have a significant effect on the amount of alcohol in a material is the amount of sugar in a material MMSTC

36. 36 Sources of Error and Weakness Incubator temperatures fluctuated slightly Yeast was left in water to rehydrate for longer than necessary Balloon did not always inflate with carbon dioxide during fermentation MMSTC

37. 37 Recap An alternative fuel source is needed to replace fossil fuels – Biofuels (ethanol) Made from fermented plant matter High alcohol content is directly related to how well a biofuel can be used MMSTC

38. 38 Recap What amount of yeast and at what temperature during fermentation will yield the greatest alcohol content? The hypothesis was that 22 °C and 3 grams of yeast would yield the highest alcohol content – Hypothesis was rejected MMSTC

39. 39 Recap Corn, water, and yeast were blended and poured into a two liter bottle with a balloon placed around the mouth The bottle was placed into an incubator to ferment Alcohol content was taken using a hydrometer MMSTC

40. 40 Recap A two factor DOE was used – Temperature (°C) – Yeast (g) Neither variable had a significant effect – Yeast can only convert the sugars that are present – Temperature speeds up the process MMSTC

41. 41 Real World Applications This information can be used by the fuel industry by saving time, money and resources used during the production of biofuel MMSTC

42. 42 Further Research Rerun the experiment using different materials such as sugar cane, beats, or switch grass. Experimenting with different plants could help find an abundant source that would be more efficient Use and compare different types of yeast MMSTC

43. 43 Acknowledgements Skip Walker – Works at local brewery MMSTC

44. 44 Works Cited "Biofuel Info." Information about Biofuel, What Is Biofuel, Renewable Energy Solutions, Ethanol Uses. Biofuel Watch, 2010. Web. 18 Sept. 2012.. Curry, Jim. "Yeast and Fermentation." MGreisMeyer. Dukes of Ale BJCP Preparation Course, 19 Apr. 2009. Web. 18 Sept. 2012.. "Dry Mill Ethanol Production." Ethanol Info. N.p., n.d. Web. 12 Oct. 2012.. MMSTC

45. Works Cited "Fermentation: World of Microbiology and Immunolgy." Fermentation. Ed. Brenda Wilmoth Learner. N.p.: n.p., n.d. N. pag.Gale Virtual Reference Library. Web. 9 Sept. 2012.. Janson, Lee W. Brew Chem 101: [the Basics of Homebrewing Chemistry]. North Adams, MA: Storey, 1996. Print. "Practical and Theoretical Considerations in the Production of High Concentrations of Alcohol by Fermentation." ScienceDirect.com. N.p., n.d. Web. 05 Dec. 2012. "Some Interesting Oil Industry Statistics." Oil Industry Statistics from Gibson Consulting. N.p., n.d. Web. 05 Dec. 2012. LaBelle -- Miley45

46. Works Cited Stairs, David. Yeast. Portland, Or.?: D. Stairs, 1999. Print. "Wet Mill Ethanol Production." Ethanol Info. N.p., n.d. Web. 12 Oct. 2012.. LaBelle -- Miley46

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