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

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

RedOx Reactions

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

ATP

Metabolic Pathways

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

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

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

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

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

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

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+

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+

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

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+

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

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?

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?)

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?

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

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

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?

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

What did we prove? • Consistent with…

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

Lab 10: Glycolysis and Fermentation