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Glycolysis 1
Lecture 23
Chapter 15
Sections 1 and 2
Chapter 14 Opener
History of Glycolysis
• 1850’s: Pasteur showed yeast can ferment sugar to
CO2 and ethanol, but he cannot replicate with
extracts, proposes “vitalism”.
• 1898: Hans and Eduard Buchner demonstrate yeast
extracts ferment sugar to CO2 and ethanol, proving
vitalism false. Birth of biochemistry.
• 1905: Harden and Young dialyze yeast extract into
two fractions:
– Retained (high molecular weight), called “zymase”.
– Dialyzed (low molecular weight), called “co-zymase”.
– Need to mix together to recover ability to ferment sugar.
– Lose activity if boil zymase, but not if boil co-zymase. Zymase
Co-zymase
Figure 2-14
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Overview of Glycolysis
• Glycolysis means “sweet splitting”
– Glyco: Greek for sweet
– Lysis: Greek for splitting or loosening
• Split a 6-carbon molecule (Glucose) to two 3-carbon
molecules (pyruvate).
• Largest carbon flux in most cells.
• Best understood metabolic pathway.
• Central to many other pathways.
• Also called fermentation:
– Yeast: Glucose -> 2 CO2 + 2 Ethanol
– Exercising muscle, RBCs, bacteria: Glucose -> 2 Lactic acid
Figure 11-4
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Glycolysis Summary
Preparatory
Stage
• 10 step breakdown of one glucose (C6) to
two pyruvates (C3).
• Balanced equation:
Glc + 2NAD+ + 2ADP + 2Pi --->
2Pyr + 2NADH + 2ATP + 2H2O + 2H+ Payoff
• Two stages: Stage
– Preparatory stage (reactions 1-5): invest 2 ATP
– Payoff stage (reactions 6-10): produce 4 ATP
Figure 15-1
Reaction 1:
Recall that hexokinase catalyzes a coupled
Hexokinase reaction:
First priming reaction Coupling the energetically unfavorable phosphorylation of
glucose to the energetically favorable hydrolysis of ATP.
Kinases: Enzymes ΔGo’
that transfer Pi from
ATP to a substrate
Hexokinase also
phosphorylates
mannose and fructose.
Page 489
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Catalyzes reversible
isomerization using
general acid-base
catalysis
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Reaction 3:
PGI Mechanism
Phosphofructokinase
Committed step, so
key regulatory site.
Second priming
reaction
Bisphosphate: 2
phosphates not linked
to one another.
Figure 15-3 Page 491
Reaction 4:
Why is phosphofructokinase the committed step in
glycolysis? Aldolase
Note change in
Glycogen Pentose-P numbering!!
Fructose-6P
Fructose-1,6-BP
Page 492
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Aldolase mechanism:
Non-enzymatic, base catalyzed aldol cleavage:
Stabilization of C-
by resonance
Ketone
Aldehyde
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Resonance stabilized
Reaction 5:
Triose phosphate isomerase (TIM)
Enediol intermediate, like PGI
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TIM has a flexible loop that closes over the active site, Toxic compound released upon dephosphorylation
preventing loss of the phosphate from the endiol intermediate of enediol intermediate
TIM barrel structure:
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Figure 15-7
Reaction 6:
GAP dehydrogenase
Preparatory
Dehydrogenases:
Stage
involved in
oxididation/reduction
reactions.
Couples favorable
oxidation to
Payoff unfavorable
Stage phosphorylation
through covalent
intermediate:
thioester
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GAP
dehydrogenase
mechanism δ-
Charge separation
δ+
δ-
Charge separation
δ+ Energy from
oxidation
conserved in
thioester
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Phosphorolysis:
Conserves energy of
thioester (originally from
oxidation) in acyl-phosphate
(Acyl-phosphate)
GAPDH catalyzes Pi
exchange
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