Carbohydrate

Metabolism: Glycolysis:

Lecturer: David Matongwe

Second year Biomedical
sciences
 Glycolysis
Definition(s)
• Glycolysis means "splitting sugars." Glucose, a six carbon
sugar, is split into two molecules of a three carbon sugar. In
the process, two molecules of ATP, two molecules of pyruvic
acid and two "high energy" electron carrying molecules of
NADH are produced.
• A set of enzymatic reactions that breakdown 6-carbon
glucose to two 3-carbon pyruvate molecules with generation
of energy
• An ATP-generating metabolic process that occurs in nearly all
living cells in which 6-glucose is converted in a series of
steps to 3-pyruvate.
• The metabolic breakdown of glucose and other sugars that
releases energy in the form of ATP.
 Biomedical Importance of Glycolysis
• Most tissues have at least some requirement for glucose.
• In brain, the requirement is substantial.
• Glycolysis, the major pathway for glucose metabolism, occurs
in the cytosol of all cells.
• It is unique in that it can function either aerobically or
anaerobically.
• In the presence of oxygen, glycolysis is the first stage of
cellular respiration. Without oxygen, glycolysis allows cells to
make small amounts of ATP. This process is called
fermentation.
• Erythrocytes, which lack mitochondria, are completely reliant
on glucose as their metabolic fuel and metabolize it by
anaerobic glycolysis.
• In addition some of the intermediates formed as a result of
glycolysis are important building blocks for other complex
 Overview of glycolysis
• From glucose to pyruvate there are 10
enzymatic steps
• The first 5 steps constitute the preparatory
phase when energy in the form of 2 ATP
molecules is spent per glucose molecule.
• The last 5 steps constitute the payoff phase
where the energy generated is used to form 4
ATP molecules on one carrier of high potential
electrons i.e. NADH per glucose molecule.
• Therefore glycolysis of a glucose molecule
leads to a net gain of 2 ATP molecules within
the cytosol
C6H12O6 + 2 NAD+ + 2 ADP + 2 P —–> 2 pyruvic

acid, (CH3(C=O)COOH + 2 ATP + 2 NADH +

2 H+
 Overview cont.
• There are 3 irreversible reactions
 The first reaction catalyzed by
hexokinase/glucokinase
 The third reaction catalyzed by
phosphofructokinase-1
 The tenth reaction catalyzed by pyruvate kinase
• Reaction catalyzed by phosphofructokinase -1 is the
most important control point in glycolysis
• The only oxidation in glycolysis is catalyzed by
glyceraldehyde 3-phosphate dehydrogenase which
forms 1,3-bisphosphoglycerate from glyceraldehyde 3-
phosphate
 Electrons are collected by NADH + H+
• Two molecules of glyceraldehyde 3-phosphate are
 GLUCOSE

Glycolysis (10 successive

reactions

Anaerobic 2 PYRUVATE Anaerobic

conditions conditions

O2
Aerobic conditions

2 ETHANOL + 2CO2 C O2
2 LACTATE

2 ACETYL-CoA
Alcoholic fermentation Anaerobic glycolysis in contracting
muscles;

O2 Lactic acid fermentation

Citric acid cycle

Animal, plant and many microbial cells
under aerobic conditions

4CO2 + 4H2O
 Glycolysis enzymatic reactions

Phase I or Preparatory Phase

• It consists of the first five steps.
i. Glucose is enzymatically phosphorylated
by ATP (first at carbon 6 and later at carbon
1) to yield fructose 1, 6-diphosphate
ii. Fructose 1, 6-diphosphate split in half to
yield 2 moles of the 3-carbon compound
glyceraldehyde 3-phosphate. Other
hexoses such as D-fructose, D-galactose
and D-mannose may also convert into
glyceraldehydes-3-phosphate.
 Phase II or Payoff Phase
• This phase represents the payoff of glycolysis,
in which the energy liberated during
conversion of 3 moles of glyceraldehydes 3-
phosphate to 2 moles of pyruvate is converted
by the coupled phosphorylation of 4 moles of
ADP to ATP.
• Although 4 moles of ATP are formed in phase
II, the net overall yield is only 2 moles of ATP
per mole of glucose oxidized, since 2 moles of
ATP are invested in phase I. The phase II is,
thus, energy conserving.
Phosphorylation of glucose (Reaction 1)

 Phosphorylation of glucose ring is the

process of adding a phosphate group to a
molecule derived from ATP resulting into 1
molecule of ATP been consumed.
 The reaction occurs with the help of the
enzyme hexokinase, an enzyme that
catalyzes the phosphorylation of many six-
membered glucose-like ring structures and
atomic magnesium (Mg) involved.
 The result of this phosphorylation is a
molecule called glucose-6-phosphate (G6P).
 Reaction of phase 1
• .

Conversion of D-glucose into glucose-6-phosphate catalyzed by the enzyme hexokinase

Conversion of G6P to F6P
 The second step of glycolysis involves
the conversion of G6P to F6P with the
help of the enzyme phosphoglucose
isomerase (PI) an isomerization
reaction.
 The reaction involves the rearrangement
of the carbon-oxygen bond to transform
the six-membered ring into a five-
membered ring.
 Reaction 2 : Phosphoglucose Isomerase

Rearrangement of G6P into F6P by glucose

phosphate isomerase (Phosphoglucose Isomerase).
 Phosphorylation of F6P
 In the third step of glycolysis,
fructose-6-phosphate is converted to
fructose- 1,6-bisphosphate (FBP).
 Similar to the reaction that occurs in
step 1 of glycolysis, a second
molecule of ATP provides the
phosphate group that is added on to
the F6P molecule.
 Reaction 3: Phosphofructokinase

Convention of fructose 6-phosphate into fructose

1,6-bisphosphate by Phosphofructokinase, with
magnesium as a cofactor.
Reaction 4: Aldolase
• Enzyme Aldolase splits fructose 1, 6-
bisphosphate into two sugars that are
isomers of each other.
• Dihydroxyacetone phosphate (DHAP)
and glyceraldehyde 3-phosphate (GAP).
 Isomerase reaction
• Triosephosphate isomerase rapidly inter-
converts the molecules dihydroxyacetone
phosphate (DHAP) to glyceraldehyde 3-
phosphate (GAP) used in next step of
Glycolysis.
• Glyceraldehyde phosphate is the only
molecule that continues in the glycolytic
pathway.
• At this point in the glycolytic pathway, we have
two 3-carbon molecules.
Reaction 5: Triosephosphate isomerase

Triosephosphate isomerase rapidly inter- converts the molecules

dihydroxyacetone phosphate (DHAP) into glyceraldehyde 3-
phosphate (GAP). GAP is used in next step of Glycolysis.
 Formation of 1,3 bisphoglycerate
 Glyceraldehyde-3-phosphate is oxidized by the
coenzyme nicotinamide adenine dinucleotide
(NAD);
 The molecule is phosphorylated by the addition of
a free phosphate group. The enzyme that
catalyzes this reaction is glyceraldehyde-3-
phosphate dehydrogenase (GAPDH).
 The enzyme GAPDH contains appropriate
structures and holds the molecule in a
conformation such that it allows the NAD molecule
to pull a hydrogen off the GAP, converting the NAD
to NADH.
 The phosphate group then attacks the GAP
molecule and releases it from the enzyme to yield
Reaction 6: Glyceraldehyde-3-phosphate
Dehydrogenase
Formation of 3-phosphoglycerate
 1,3 bisphoglycerate is converted to 3-
phosphoglycerate by the enzyme phosphoglycerate
kinase (PGK). (loss of a phosphate group from the
starting material). The phosphate is transferred to a
molecule of ADP that yields our first molecule of ATP.

 Note: Two molecules of ATP synthesized at this step cancel

the first two molecules of ATP that we used, leaving us with
a net of 0 ATP molecules up to this stage of glycolysis

 Magnesium is involved to shield the negative charges

on the phosphate groups of the ATP molecule.
Reaction 7: Phosphoglycerate Kinase

Phosphoglycerate kinase transfers a phosphate group

from 1,3-bisphosphoglycerate to ADP to form ATP and
3-phosphoglycerate.
Formation of 2-phosphoglycerate
 This step involves a simple rearrangement of
the position of the phosphate group on the 3
phosphoglycerate molecule, making it 2
phosphoglycerate by an enzyme
phosphoglycerate mutase (PGM).
 A mutase is an enzyme that catalyzes the
transfer of a functional group from one position
on a molecule to another.
Reaction 8: Phosphoglycerate Mutase

The P from 3- phosphoglycerate from the 3rd carbon is relocated

to the 2nd carbon to form 2-phosphoglycerate by phosphoglycero
mutase .
Formation of phosphoenolpyruvate

 The conversion of 2 phosphoglycerate to

phosphoenolpyruvate (PEP).
 The reaction is catalyzed by the enzyme
enolase.
 Enolase works by removing a water group,
or dehydrating the 2 phosphoglycerate.
 The specificity of the enzyme pocket allows for
the reaction to occur through a series of steps
which too complicated
Reaction 9: Enolase
Formation of pyruvic acid
 The enzyme pyruvate kinase transfers a P
from phosphoenolpyruvate (PEP) to ADP to
form pyruvic acid and ATP.
 Final step of glycolysis where
phosphoenolpyruvate is converted into
pyruvate with the help of the enzyme pyruvate
kinase.
 Reaction involves the transfer of a phosphate
group attached to the 2′ carbon of the PEP to a
molecule of ADP, yielding ATP.
Note: Two molecules of PEP resulting into 2 ATP
molecules being generated.
Step 10: Pyruvate Kinase

The enzyme pyruvate kinase transfers a P from

phosphoenolpyruvate (PEP) to ADP to form pyruvic acid and
ATP is formed.
Steps 1 and 3 = – 2ATP
Steps 7 and 10 = + 4 ATP
Net “visible” ATP produced = 2.
 Anaerobic Glycolysis

• Pyruvate an end product of glycolysis has several

separate fates depending on the microcellular
environment (specifically, oxygen availability, energy
demand, and the presence or absence of mitochondria).
• In mitochondria-containing cells, pyruvate can enter the
citric acid cycle within the mitochondrial matrix and
undergo oxidative phosphorylation. Aptly named due to
its dependence on oxygen as the final electron
acceptor, oxidative phosphorylation cannot take place in
the absence of oxygen.
• Moreover, as the enzymes of both the citric acid cycle
and electron transport chain are within the mitochondria,
cells lacking mitochondria (e.g., erythrocytes) cannot
rely on oxidative phosphorylation for energy production.
• In erythrocytes and oxygen-deprived tissue,
pyruvate remains within the cytoplasm and
converts to lactate, a process referred to
as anaerobic glycolysis.
• This final reaction allows for the regeneration of
NAD+, a cofactor that must be available in high
enough intracellular concentrations for the earlier
reactions of glycolysis to remain favorable.
• Compared to oxidative phosphorylation, however,
anaerobic glycolysis is significantly less efficient,
providing a net production of only 2 ATP per
glucose molecule (versus 32 ATP per glucose
molecule produced during oxidative
phosphorylation).
• ATP Yield:
• For every glucose molecule, cellular
respiration produces:
– Glycolysis: 2 ATP
– Krebs Cycle (Citric Acid Cycle): 2 ATP
– Oxidative Phosphorylation: 28 ATP
– Fermentation: 2 ATP
• Therefore, the net ATP production per glucose
molecule is 32 ATP.
 Overview of Respiration

Organic Compounds

Glycolysis
Oxygen Present Oxygen Absent

Cellular Respiration Fermentation

(aerobic) (anaerobic)
 Types of fermentation

Fermentation

Lactic acid Alcohol

 Fermentation (cont)
• Lactic Acid Fermentation occurs in
muscle cells
• In the cytosolic enzyme lactate
dehydrogenase converts pyruvate to
lactic acid
• lactic acid results in muscle fatigue
and cramps
• produces NAD+ from NADH that goes
back to glycolysis
Lactic Acid Fermentation
 Fermentation (cont)
• Alcoholic fermentation produces
alcohol (ethyl) and carbon dioxide

• NAD+ that goes back to glycolysis

Alcoholic Fermentation
 Importance of Fermentation
• Allows glycolysis to continue

• Used in the production of food

stuffs such as bread, cheese, and
yogurt.
The end