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 CH 4 (methane) C 2 H 5 OH H 3 C-CH 2 -OH (ethanol) C 8 H 18 CH 3 (CH 2 ) 6 CH 3 (octane) H 2 O CO 2 Similarities in components/structure? Differences in behavior? Differences in structure?

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

5. RedOx Reactions

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

7. ATP

11. Metabolic Pathways

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

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

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

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

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

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

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

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

25. Sugar/CO 2 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?) –CO 2 (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?) –CO 2 (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. no heating parafilm!!! Thermometer should measure GAS temp, leave it up high! AFTER

33. What did we prove? Consistent with…

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