Download - Lab 10: Glycolysis and Fermentation
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Lab 10: Glycolysis and Fermentation
What’s so sweet about Sugar?
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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
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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?
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So where are ‘calories’?
Energy (heat) needed to raise 1g water, 1oC
Saturated Fat
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RedOx Reactions
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OIL RIG
OIL = Oxidation Is Loss
RIG = Reduction Is Gainof electrons
of electrons
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ATP
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ATP
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ATP
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ATP
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Metabolic Pathways
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Metabolic Pathways
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Metabolic Pathways
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Glycolysis• Glyco – glucose/sugar
• Lysis – “to loosen”, take apart
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Glycolysis
Freeman, Scott. Biological Science. 4th Edition.
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Glycolysis + fermentation
Freeman, Scott. Biological Science. 4th Edition.
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Glycolysis + Fermentation
Freeman, Scott. Biological Science. 4th Edition.
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Here’s the claim:Glycolysis –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
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Here’s the claim:Glycolysis –
Fermentation –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
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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+
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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+
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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+
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Standard Protocol
• Pg. 10-7 of lab manual
• Change to 500 mL
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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+
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Sugar/CO2 Presence?
• Appendix C
• ***READ APPENDIX C*** – REALLY (lots of helpful stuff…in the text)
• What about ETOH?
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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?
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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?)
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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?
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After Mixing
• What will your solution look like?– “murky”?
• How do you speed it up?
• What conditions do we WANT?
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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
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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?
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no heating parafilm!!!
Thermometer should measure GAS temp, leave it up high!
AFTER
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What did we prove?
• Consistent with…
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Next Week• Assess: Glycolysis and fermentation
• Lab report: Fermentation with a Bullet– One/group in Dropbox
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