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Lab 10: Glycolysis and Fermentation

Lab 10: Glycolysis and Fermentation. What’s so sweet about Sugar?. Objectives. Discuss ATP and where the ‘energy’ is Describe what fermentation is and where the ‘energy’ is Design an experiment to ‘test’ fermentation Recognize/explain what happens in a RedOx reaction. Thinking it Through.

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Lab 10: Glycolysis and Fermentation

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  1. Lab 10: Glycolysis and Fermentation What’s so sweet about Sugar?

  2. Objectives • Discuss ATP and where the ‘energy’ is • Describe what fermentation is and where the ‘energy’ is • Design an experiment to ‘test’ fermentation • Recognize/explain what happens in a RedOx reaction

  3. Thinking it Through • CH4 (methane) C2H5OH H3C-CH2-OH (ethanol) C8H18 CH3(CH2)6CH3 (octane) • H2O CO2 • Similarities in components/structure? • Differences in behavior? • Differences in structure?

  4. So where are ‘calories’? Energy (heat) needed to raise 1g water, 1oC Saturated Fat

  5. RedOx Reactions

  6. OIL RIG OIL = Oxidation Is Loss of electrons RIG = Reduction Is Gain of electrons

  7. ATP

  8. ATP

  9. ATP

  10. ATP

  11. Metabolic Pathways

  12. Metabolic Pathways

  13. Metabolic Pathways

  14. Glycolysis • Glyco – glucose/sugar • Lysis – “to loosen”, take apart

  15. Glycolysis Freeman, Scott. Biological Science. 4th Edition.

  16. Glycolysis + fermentation Freeman, Scott. Biological Science. 4th Edition.

  17. Glycolysis + Fermentation Freeman, Scott. Biological Science. 4th Edition.

  18. Here’s the claim: Glycolysis – glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP

  19. Here’s the claim: Glycolysis – Fermentation – glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+

  20. Here’s the claim: Glycolysis – Fermentation – What can you test? Rank in terms of feasibility, safety, etc. glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+

  21. Here’s the claim: Glycolysis – Fermentation – What can you test? Rank in terms of feasibility, safety, etc. glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+

  22. Here’s the claim: Glycolysis – Fermentation – What can you test? Rank in terms of feasibility, safety, etc. What will glyco + ferment? How???? glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+

  23. Standard Protocol • Pg. 10-7 of lab manual • Change to 500 mL

  24. Here’s the claim: Glycolysis – Fermentation – What can you test? Rank in terms of feasibility, safety, etc. What will glyco + ferment? How/when to do CO2, sugar, ETOH? glucose + 2ADP + 2Pi+ 2NAD+ => 2 pyruvic acid + 2NADH++ 2ATP pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+

  25. Sugar/CO2 Presence? • Appendix C • ***READ APPENDIX C*** – REALLY (lots of helpful stuff…in the text) • What about ETOH?

  26. Group discussion • On what basis will you be drawing conclusions at the end of the experiment • making comparisons between the final result and...? • What if we wanted to produce a lot of ETOH, or very little glucose?

  27. Optimizations • Concept: limiting components. How do you max your yield (sensitivity) for: • Glucose (how might you limit in final solution?) • CO2 (Maximize) • ETOH (Maximize) • Use standard protocol (10-7) as a baseline and adjust amounts from there per your role (could there be too much bread?)

  28. Optimizations • Concept: limiting components. How do you max your yield (sensitivity) for: • Glucose (how might you limit in final solution?) • CO2 (Maximize) • ETOH (Maximize) • Use standard protocol (10-7) as a baseline and adjust amounts from there per your role (could there be too much bread?) • What do we do? Glucose groups – 1X or 2X?

  29. After Mixing • What will your solution look like? • “murky”? • How do you speed it up? • What conditions do we WANT?

  30. First part • 1) Yeast mixed with glucose solution • 2) Take glucose pre-sample (in plastic tube) and place on ice – how much? Answer: see ‘candidate solution’ in protocol • 3) Hook up BaOH glass tube to flask and follow protocol • 4) Cook after sealing/taping • Turn on hot plates

  31. After Cooking • 1) BaOH outcome? • 2) Take another yeast solution extract for sugar testing – how many do we have? Why? • How to get chunks out? • Follow Benedict’s protocol • 3) Heat the beer…get ETOH – how to do so? • How to test?

  32. AFTER no heating parafilm!!! Thermometer should measure GAS temp, leave it up high!

  33. What did we prove? • Consistent with…

  34. Next Week • Assess: Glycolysis and fermentation • Lab report: Fermentation with a Bullet • One/group in Dropbox

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