Glycolysis and Fate of

pyruvate
Dr. Amar Babikir Elhussein
BSc, MSc &PhD
 Glycolysis

• 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.

• Glycolysis is both the principal route for glucose metabolism and the
main pathway for the metabolism of fructose, galactose, and other
carbohydrates derived from the diet.
• All enzymes of glycolysis are found in the cytosol.

• Most tissues have at least some requirement for glucose.

• In brain, the requirement for glucose is substantial.

• Erythrocytes, which lack mitochondria, are completely depend on

glucose as metabolic fuel and metabolize it in anaerobic glycolysis.
• The ability of glycolysis to provide ATP in the absence of oxygen is
important, because it allows skeletal muscles to perform at very high
levels when oxygen supply is insufficient.

• Certain disease are shown when enzymes of glycolysis are deficient

(Heamolytic anaemias).
❑ Reactions of Glycolysis:

• The overall equation for glycolysis from glucose to lactate is as

follows:

• Glucose + 2ADP +2Pi 2 Lactate +2 ATP +2H2O.

• Glucose enters glycolysis by phosphorylation to glucose 6-phosphate,

catalyzed by hexokinase, using ATP as the phosphate donor.

• Under physiologic conditions, the phosphorylation of glucose to

glucose 6-phosphate can be regarded as irreversible.

• Hexokinase is inhibited allosterically by its product, glucose 6-

phosphate.
 Hexokinase Glucokinase
In most tissues. In liver parenchymal cells and β
cells of the pancreas.
Able to phosphorylate several Phosphorylation of only glucose.
hexoses in addition to glucose.
Hexokinase has a low Km It has a much higher Km,
(and, therefore, a high requiring a higher glucose
affinity). concentration for half-saturation.
Constitutive Adaptive
• Glucose 6-phosphate is then phosphorylated with ATP catalyzed by the enzyme

phosphofructokinase (phosphofructokinase-1), forming fructose 1,6-

bisphosphate.

• The phosphofructokinase reaction is irreversible under physiologic conditions;

it is both inducible and subject to allosteric regulation and has a major role in

regulating the rate of glycolysis.

• Fructose 1,6-bisphosphate is cleaved by aldolase (fructose 1,6-bisphosphate

aldolase) into two triose phosphates, glyceraldehyde 3-phosphate and

dihydroxyacetone phosphate.

• Glyceraldehyde 3-phosphate and dihydroxyacetone phosphate are

interconverted by the enzyme phosphotriose isomerase.

• Glycolysis continues with the oxidation of glyceraldehyde 3-phosphate to 1,3-

bisphosphoglycerate.

• The enzyme catalyzing this oxidation, glyceraldehyde 3-phosphate

dehydrogenase, is NAD-dependent.

• The reaction, catalyzed by phosphoglycerate kinase, phosphate is transferred

from 1,3-bisphosphoglycerate onto ADP, forming ATP (substrate-level

phosphorylation) and 3-phosphoglycerate.

• 3-Phosphoglycerate is isomerized to 2-phosphoglycerate by

phosphoglycerate mutase.

✓NB: Since two molecules of triose phosphate are formed per molecule of

glucose, two molecules of ATP are generated at this stage per molecule of

glucose undergoing glycolysis.

• This step may competitively inhibited by arsenic.

• The subsequent step is catalyzed by enolase and involves a dehydration,

forming phosphoenolpyruvate.

• Enolase is inhibited by fluoride.

• The enzyme is also dependent on the presence of either Mg2+ or Mn2+.

• The phosphate of phosphoenolpyruvate is transferred to ADP by pyruvate

kinase to generate, at this stage, two molecules of ATP per molecule of glucose

oxidized.
• The product of the enzyme-catalyzed reaction, enolpyruvate, undergoes

spontaneous (nonenzymic) isomerization to pyruvate.

• Under anaerobic conditions, the re-oxidation of NADH through the

respiratory chain to oxygen is prevented.

• Pyruvate is reduced by the NADH to lactate, the reaction being catalyzed by

lactate dehydrogenase.
• The re-oxidation of NADH via lactate formation allows glycolysis to proceed in

the absence of oxygen by regenerating sufficient NAD + for another cycle of the

reaction catalyzed by glyceraldehyde-3-phosphate dehydrogenase.

• Under aerobic conditions, pyruvate is taken up into mitochondria and after

conversion to acetyl-CoA is oxidized to CO2 by the citric acid cycle.

• The reducing equivalents from the NADH + H+ formed in glycolysis are taken

up into mitochondria for oxidation.

Summary of anaerobic glycolysis.
Reactions involving the production
or consumption of ATP or NADH
are indicated.
The three irreversible
reactions of glycolysis are shown
with thick arrows.
DHAP =
dihydroxyacetone phosphate.
• Glycolysis in erythrocytes, even under aerobic conditions, always terminates

in lactate, because the subsequent reactions of pyruvate are mitochondrial, and

erythrocytes lack mitochondria.

• Other tissues that normally derive much of their energy from glycolysis and

produce lactate include brain, gastrointestinal tract, renal medulla, retina, and

skin.
❑Regulation of glycolysis:

• Most of the reactions of glycolysis are reversible, except three

reactions.

• These reactions, catalyzed by hexokinase (and glucokinase),

phosphofructokinase, and pyruvate kinase, are the major sites of
regulation of glycolysis.
The regulation of glycolysis by allosteric activation or

inhibition, or the phosphorylation/dephosphorylation of

rate-limiting enzymes, is shortterm “that is, they

influence glucose consumption over periods of minutes

or hours.

Superimposed on these moment-to-moment effects are

slower, and often more profound, hormonal influences

on the amount of enzyme protein synthesized.

• These effects can result in 10-fold to 20- fold increases in enzyme activity that

typically occur over hours to days.

• Regular consumption of meals rich in carbohydrate or administration of insulin

initiates an increase in the amount of glucokinase, phosphofructokinase, and

pyruvate kinase in liver .

• These changes reflect an increase in gene transcription, resulting in increased

enzyme synthesis.
• High activity of these three enzymes favors the conversion of glucose to

pyruvate, a characteristic of the well-fed state .

• Conversely, gene transcription and synthesis of glucokinase,

phosphofructokinase, and pyruvate kinase are decreased when plasma glucagon

is high and insulin is low, for example, as seen in fasting or diabetes.

In the erythrocytes of many mammals, the reaction catalyzed by

phosphoglycerate kinase may be bypassed by phosphate of 1,3-

bisphosphoglycerate.

Bisphosphoglycerate mutase catalyzes the conversion of 1,3

bisphosphoglycerate to 2,3-bisphosphoglycerate, which is converted to 3-

phosphoglycerate by 2,3- bisphosphoglycerate phosphatase.

This process provide 2,3-

bisphosphoglycerate, which binds

to hemoglobin, decreasing its

affinity for oxygen and so making

oxygen more readily available to

tissue.
❑Oxidation of pyruvate to acetyl COA:

• Pyruvate, formed in the cytosol, is transported into the mitochondria.

• Inside the mitochondrion, pyruvate is oxidatively decarboxylated to acetyl-

CoA by a multienzyme complex (pyruvate dehydrogenase complex) that is

associated with the inner mitochondrial membrane.

• Pyruvate is decarboxylated by the pyruvate dehydrogenase to a

hydroxyethyl derivative of the thiazole ring of enzyme bound thiamin

diphosphate, which in turn reacts with oxidized lipoamide, the prosthetic

group of dihydrolipoyl transacetylase, to form acetyl lipoamide.

• Thiamin is vitamin B1
Mechanism of action of the pyruvate dehydrogenase complex.
TPP = thiamine pyrophosphate; L = lipoic acid.
• Pyruvate Dehydrogenase Is Regulated by End-Product Inhibition & Covalent
Modification.

• Pyruvate dehydrogenase is inhibited by its products, acetyl-CoA and NADH.

• It is also regulated by phosphorylation by a kinase of three serine residues on the

pyruvate dehydrogenase component of the multienzyme complex, resulting in
decreased activity.

• The kinase is activated by increases in the [ATP]/[ADP], [acetyl-CoA]/[CoA], and

[NADH]/[NAD+] ratios.

• Thus, pyruvate dehydrogenase— and therefore glycolysis—is inhibited not only by a

high-energy potential but also when fatty acids are being oxidized.
• Thus, in starvation, when free fatty acid concentrations increase, there is a

decrease in the proportion of the enzyme in the active form, leading to a

sparing of carbohydrate.

• In adipose tissue, where glucose provides acetyl CoA for lipogenesis, the

enzyme is activated in response to insulin.

❑CLINICAL ASPECTS:

• Arsenite and mercuric ions react with the - SH groups of lipoic acid and

inhibit pyruvate dehydrogenase, as does a dietary deficiency of thiamin,

allowing pyruvate to accumulate.

• Nutritionally deprived alcoholics are thiamin-deficient and may develop

potentially fatal pyruvic and lactic acidosis.

• Patients with inherited pyruvate dehydrogenase deficiency, which can be due to

defects in one or more of the components of the enzyme complex, also present with

lactic acidosis.

• Because of its dependence on glucose as a fuel, brain is a prominent tissue where

these metabolic defects manifest themselves in neurologic disturbances.

• Inherited aldolase A deficiency and pyruvate kinase deficiency in erythrocytes cause

hemolytic anemia.
• The exercise capacity of patients with muscle phosphofructokinase

deficiency is low, particularly on high-carbohydrate diets.

• By providing an alternative lipid fuel, eg, during starvation, when blood free

fatty acids and ketone bodies are increased, work capacity is improved.
• Reference:
✓Harper`s Biochemistry
✓Lippincott's Illustrated Reviews Biochemistry

Thank you