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
ATP
ATP
ATP
Metabolic Pathways
Metabolic Pathways
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 –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
Fermentation –
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
Here’s the claim:
Glycolysis –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
Fermentation –
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
What can you test?
Rank in terms of feasibility, safety, etc.
Here’s the claim:
Glycolysis –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
Fermentation –
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
What can you test?
Rank in terms of feasibility, safety, etc.
Here’s the claim:
Glycolysis –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
Fermentation –
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
What can you test?
Rank in terms of feasibility, safety, etc.
What will glyco + ferment?
How????
Standard Protocol
• Pg. 10-7 of lab manual
• Change to 500 mL
Here’s the claim:
Glycolysis –
glucose + 2ADP + 2Pi + 2NAD+ => 2 pyruvic acid + 2NADH+ + 2ATP
Fermentation –
pyruvic acid + NADH+ => ethanol + CO2 + 2NAD+
What can you test?
Rank in terms of feasibility, safety, etc.
What will glyco + ferment?
How/when to do CO2, sugar, ETOH?
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