P.10.

GLYCOLYSIS AND THE ▪ reciprocal actions ensure that both

OXIDATION OF PYRUVATE (PART 2) pathways are not fully active at the
Dr.Salango | February 8, 2017 same time
▪ During well-fed state:
elevated insulin, dec. Glucagon
OUTLINE =increase F-2,6-BP
I. The Two Phases of Glycolysis ▪ During fasting:
A. Preparative Phase (Continuation) elevated glucagon, dec. Insulin
B. ATP generating Phase =decrease F-2,6-BP
II. Anaerobic Glycolysis
III. Oxidative Decarboxylation of Pyruvate D. CLEAVAGE OF FRUCTOSE-1,6-BIPHOSPHATE
IV. Other Sugars Feed into Glycolysis
V. Regulation of Glycolysis
VI. Enzyme Complexes Facilitating Channels

A.PREPARATIVE PHASE (Continuation)

C. PHOSPHORYLATION OF FRUCTOSE-6-PHOSPHATE
• Hydrolysis of F-1,6-BP into two 3-carbon products:
o Dihydroxyacetone phosphate (DHAP)
o Glyceraldehyde 3-phosphate (G3P)
• Enzyme: Aldolase
o Aldolase B: isoform of aldolase found primarily
in the liver, also cleaves fructose-1-phosphate
• Hexokinase reaction and functions in the metabolism of dietary
• Not readily reversible; most important control point fructose
and the rate-limiting and committed step • Reversible in three way and not regulated reaction
• Ratio of fructose-6-phosphate to glucose-6-phosphate is • Ratio is 50:50
40:60 E. ISOMERIZATION OF DIHYDROXYACETONE
• Phosphorylation of fructose-6-phosphate (F-6-P) to PHOSPHATE
fructose-1,6-biphosphate (F-1,6-BP)
o By utilizing a second ATP
o Catalyzed by the enzyme Phosphofructokinase-
1/PFK-1/
6-Phosphofructo-1-Kinase/
Fructose 1-phosphorylase • The two products of the aldolase reaction equilibrate
• PFK-1 is controlled by the available concentrations of
readily in a reaction catalyzed by:
the substrates ATP and fructose-6-phosphate o Enzyme: Triose phosphate isomerase
o PFK-1 is inhibited allosterically by elevated
(phosphotriose isomerase):
levels of ATP ▪ interconverts dihydroxyacetone
o Fructose-6-phosphate forms fructose-2,6-
phosphate (DHAP) and
biphosphate by the
glyceraldehyde-3-phosphate (G3P)
enzymephosphofructokinase-2 (PFK-2)
o DHAP is utilized in triacylglycerol synthesis
o PFK-2: enzyme that converts to opposite
• Net production of two molecules of G3P
reaction; has a kinase activity that produces
• Succeeding reactions of glycolysis utilize G3P as a
fructose-2,6-biphosphate and a phosphatase
substrate.
activity that dephosphorylates fructose-2,6-
• Bidirectional
biphosphate back to fructose 6-phosphate
• PFK-1 is active in Glycolysis; inactive in gluconeogenesis
A.ATP GENERATING PHASE
PFK-2 is active in Gluconeogenesis; inactive in glycolysis
(rate limiting enzyme in gluconeogenesis)
F. OXIDATION OF GLYCERALDEHYDE-3-PHOSPHATE
o Other name: Fructose -1,6 Biphosphatase
• Fructose-2,6-bisphosphate (F-2,6-BP):
▪ most potent activator of PFK-1
▪ inhibitor of fructose 1,6-
bisphosphatase aka PKF-2(enzyme of
gluconeogenesis)
▪ intracellular signal, indicating
abundance of glucose

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• Catalyzes the NAD+ dependent oxidation of G3P
o to 1,3-biphosphoglycerate (1,3-BPG)
o Enzyme: glyceraldehyde-3-phosphate
dehydrogenase
o NADH formed by this reaction must be
reoxidized thru:
▪ NADH linked conversion of pyruvate
to lactate (anaerobic)
▪ Oxidation of NADH via respiratory
chain (aerobic) which requires malate-
aspartate and G-3-P substrate
shuttles
• Reversible
• In RBCs, the first site in glycolysis for the generation of
ATP may be bypassed, leading to the formation of 2,3-
biphosphoglycerate, which is important in decreasing
the affinity of Hgb for O2 (Oxyhemoglobin dissociation
curve shifts to the right)
G. SYNTHESIS OF PHOSPHOGLYCERATE PRODUCING ATP
• The high-energy phosphate pf 1,3-BPG is used to forom
ATP and 3-phosphoglycerate (3PG
o Enzyme: phosphoglycerate kinase
▪ Physiologically reversible
▪ Replaces the two ATP molecules
consumed by earlier reaction
o Used to synthesize ATP from ADP
• Substrate level phosphorylation
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H. SHIFT TO PHOSPHATE GROUP
• Conversion of 3PG to 2PG
o Enzyme: Phosphoglycerate mutase
• Freely reversible
• Aimed at converting a relatively low energy
phosphoacyl-ester of 3PG to a high-energy to form and
harvesting the phosphate as ATP
I. DEHYDRATION OF 2-PHOSPHOGLYCERATE
• Dehydration/ conversion of 2-phosphoglycerate to form
phosphoenolpyruvate (PEP)
o Enzyme: enolase
o PEP contains a high energy enol phosphate
o Redistributes the energy within the substrate
• Reversible
• Fluoride: inhibits enolase
J. FORMATION OF PYRUVATE, PRODUCING ATP
• Conversion of PEP to pyruvate
o Enzyme:pyruvate kinase (PK)
• Third irreversible reaction of glycolysis
• Substrate level phosphorylation
• Final reaction kinase
• Strongly exergonic reaction, high energy enol phosphate
in PEP is used to synthesize ATP from ADP
• Loss of phosphate by PEP leads to the production of
pyruvate in an unstable enol form which spontaneously
tautomerizes to the more stable, keto form of pyruvate.
This reaction contributes a large proportion of the free
energy of hydrolysis
• PRODUCTS:
o 4 ATP from one glucose (from 2 G3P)
o 2 pyruvates
o 2 NADH
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Summary

II. ANAEROBIC GLYCOLYSIS

• Two molecules of ATP are generated for each molecule
of glucose converted to two molecules of lactate. There
is no net production or consumption of NADH
• In the absence of oxygen, pyruvate is reduced by NADH
to lactic acid
o Enzyme: Lactate Dehydrogenase
o Conditions of stress like fight or flight activities:
anaerobic glycolysis
▪ e.g. 100-meter dash, running as fast

as you can
• Without Oa- oxidative respiration
• With O2, reaction is relatively unstable so it find its way
to something that stabilizes it
o H ion will bind (usual source: NADH
o Final product: Lactic Acid
• Lactic Acid is not at all bad,
o when accumulates, it directly goes to the
blood stream, then to the liver where it will be
converted back to glucose.
• In bacteria and lower forms of animals, the product of
Lactic Acid is ethyl alcohol and CO2
• Tissues that normally derive their energy from glycolysis
and glucose lactate:
o Skeletal muscle
o Brain
o GIT

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 IV. OTHER SUGRAS FEED INTO GLYCOLYSIS

• Dehydrogenase reaction: removal of hydrogen making it

more oxygen containing than hydrogen containing 
less electron  more oxidized  more unstable more
accepting of phosphate groups
• Glycolysis: oxidation of glucose
❖ DHAP: to prevent concentration gradient inhibition
o Too much G3P: other reactions will not
proceed
o “spillage control”
o Shuttle mechanism: tranport of other
molecules inside mitochondria

III. OXIDATIVE DECARBOXYLATION OF PYRUVATE

• Under normal situation: aerobic is more dominant

• The crucial element in aerobic glycolysis is the
conversion of pyruvate to acetyl CoA
o Enzyme: Pyruvate dehydrogenase
• Acetyl CoA- a major fuel for the TCA cycle and the V. REGULATION OF GLYCOLYSIS
building block for fatty acid synthesis • Feedback inhibition system
o Blocked when there is fatty acid in the • ATP plays a crucial role in regulation of glycolysis:
cytoplasm: “Randle effect” o ↑ATP:
o Stored ATP→anaerobic glycolysis→aerobic Negative feedback inhibition on conversion of
glycolysis→fatty acid fructose-6-phosphate to fructose-1,6-
• Irreversible biphosphate
• Important pathway in tissues with a high oxidative ▪ PFK-1: inhibited by ATP
capacity such as cardiac muscle o ↑ADP:
stimulates PFK-1

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 VI. ENZYME COMPLEXES FACILITATING CHANNELS

CHECKPOINT
T/F
1. The phosphorylation of glucose-6-phosphate to glucose
by the action of glucokinase uses ATP and is reversible.
2. All reactions involved in glycolysis are reversible, thus
maintaining homeostasis in the body’s blood sugar.
3. Pyruvate kinase is a final reaction kinase.
4. PFK-1 is inactive in gluconeogeneis; active in
glycogenolysis
5. Lactic acid is a waste product of anaerobic metabolism
and cannot be converted to glucose, thus it is excreted
out of the body.

ANSWERS: F F T F F

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 SUMMARY OF REACTIONS

Substrate Enzyme Product Reaction COMMENTS

PREPARATIVE PHASE

Glucose Glucokinase Glucose-6-Phosphate Phosphorylation Uses ATP

Irreversible

Glucose-6-Phosphate Phosphohexose isomerase Fructose-6-Phosphate Aldose-Ketose isomerization Reversible

Fructose-6-Phosphate PFK-1 Fructose-1,6-Biphosphate Phosphorylation Uses ATP

Irreversible

Fructose-1,6- Aldolase Dihydroxyacetone phosphate Hydrolysis Reversible

Biphosphate
Glyceraldehyde-3-phosphate

Dihydroxyacetone Triose phosphate Glyceraldehyde-3-phosphate Isomerization Reversible

phosphate isomerase

ATP GENERATING PHASE

Glyceraldehyde-3- Glyceraldehyde-3- 1,3-biphosphoglycerate Oxidation NAD dependent

phosphate phosphate dehydrogenase
Reversible

1,3-biphosphoglycerate Phosphoglycerate kinase 3-phosphoglycerate Substrate level Reversible

phosphorylation

3-phosphoglycerate Phosphoglycerate mutase 2-phosphoglycerate Shifting Reversible

2-phosphoglycerate Enolase Phosphoenolpyruvate Dehydration Reversible

Phosphoenolpyruvate Pyruvate kinase Pyruvate Substrate level Irreversible

phosphorylation

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