The sequence of reactions from glucose to pyruvate is similar in most organisms and most

types of cells.
In contrast, the fate of pyruvate is variable.

Pyruvate is most commonly metabolized in one of three ways, depending on the type of
organism & the presence or absence of O .

1- Aerobic Condition
• AEROBIC RESPIRATION
• is the release of energy from
the breakdown of glucose by
combining it with oxygen
inside living cells.

3- Anaerobic conditions:
2- Anaerobic conditions (microorganism and yeast):
(muscles, RBCs): • ALCOHOLIC FERMENTATION
• LACTIC ACID FERMENTATION • Conversion of pyruvate to
• Conversion of pyruvate to ethanol
lactate

A key to understand the biochemical logic behind two of these reactions of pyruvate (Anaerobic conditions)
is to recognize that glycolysis needs a continuing supply of NAD+. (HOW?)

>> If NO oxygen is present to reoxidize NADH to NAD+ then another way must be found to reoxidize it.
 ​Diverse fates of
​Pyruvate

​Oxidation of pyruvate to/acetyl-CoA,

​which enters the citric acid cycle for
​further oxidation

​When glycolysis is complete (in the

​cytosol), two pyruvate molecules are
​left.

​Under aerobic conditions, pyruvate

​undergoes several changes to become
​acetyl Coenzyme A (acetyl CoA) in the
​1- Aerobic condition
​mitochondrial matrix (in eukaryotes)

​Once in the mitochondrion, aerobic

​respiration would break them down
​further, releasing more energy.

​Citric acid cycle (Krebs cycle)

​This occurs in two major stages:
​Electron transport system

​(muscles, RBCs)

​When we race too fast or work too

​hard, the oxygen supply can't keep up
​with the need >>> That is when our
​muscles switch from aerobic respiration
​to lactic acid fermentation.

​Lactic acid fermentation is the process

​2- Anaerobic condition/ ​by which our muscle cells deal with
​Lactic acid fermentation ​pyruvate during anaerobic respiration
​and produce lactic acid.

​The regeneration of NAD in the

​reduction of pyruvate to lactate
​sustains the continued operation of
​glycolysis under anaerobic conditions.

​lactic acid fermentation also takes

​place in a variety of microorganisms.

​(microorganisms, yeast)

​Ethanol is formed from pyruvate in

​yeast and several other microorganisms.

​Decarboxylation of pyruvate to
​acetaldehyde by Pyruvate
​3- Anaerobic condition/
​decarboxylase. >>This enzyme require
​Mg* and the cofactor, thiamine ​Anabolic alcoholic
​Two reactions lead to the production
​pyrophosphate (TPP) ​fermentation
​of ethanol:
​Reduction of acetaldehyde to ethanol
​by Alcohol dehydrogenase

​ The regeneration of NAD in the

​reduction of pyruvate to ethanol
​sustains the continued operation of
​glycolysis under anaerobic conditions.
 Anaerobic Condition

Summary
• Pyruvate is converted to lactate in anaerobic tissues, such as actively
metabolizing muscle. NAD+ is recycled in the process
• In some organisms, pyruvate is converted to ethanol in a process requiring
Mg2+ and thiamine pyrophosphate as a coenzyme
• NAD+ Needs to be Recycled to Prevent Decrease in Oxidation
Reactions, So NAD + will be present for further glycolysis to take place
Metabolic regulation of Glycolysis
• The biochemical pathways are composed of groups of coordinated enzymes that perform a specific
metabolic pathway.

• In general, these enzyme groups are composed of many enzymes, but only one or more of these
enzymes along a given pathway have the capacity to affect the flux of the pathway. These
enzymes known as REGULATORY ENZYMES.

• REGULATORY ENZYMES mostly found at or near the initial steps in a pathway, or part of a
branch point or cross-over point between pathways (where a metabolite can be potentially
converted into several products in different pathways)

• Regulatory Enzymes Regulation : Of the 10 steps in the glycolytic pathway, three involve large
negative G and are essentially irreversible, These are;
◦ steps1: (phosphorylation of glucose) catalysed by hexokinase (or glucokinase).
◦ step 3: (phosphorylation of fructose-6-phosphate) catalysed by phosphofructokinase.
◦ step 10: (transfer of phosphate from phosphoenolpyruvate to ADP) catalysed by pyruvate
kinase
 ​Hexokinases, First step
​Pyruvate Kinases, Last step

​the enzyme catalyzing the first step of

​glycolysis. ​the enzyme catalyzing the last step of
​glycolysis

​>> High concentrations of this molecule

​It is regulated by allosteric effector ( ​allosteric effectors
​signal that the cell no longer requires glucose
​feedback inhibition)>> it is inhibited by its
​for energy, instead the cells need to store ​It is regulated by
​product, glucose 6-phosphate.
​glucose in the form of glycogen. ​covalent modification.

​Metabolic regulation
​of Glycolysis
​A high level of ATP inhibits the enzyme by
​AMP diminishes and citrate enhances the
​Allosteric inhibitor ​ATP & Citrate ​decreasing its affinity for substrate fructose
​inhibitory effect of ATP
​It is highly regulated with various allosteric ​6-phosphate.
​inhibitors and activators:
​Allosteric activator ​Fructose 2,6 bisphosphate & ADP and AMP ​Glycolysis is stimulated as the Energy
​Charge falls.

​the enzyme catalyzing the third step of

​glycolysis: is the most important control
​element in the mammalian glycolytic pathway

​Phosphofructokinases, Third step

 Energy yield from glycolysis
The ATP yield from glycolysis is different in anaerobic and aerobic conditions

During aerobic (oxygen plenty) During anaerobic (oxygen de ciency)

condition “Oxygen is available” condition
• The two NADH generated in step 6 can • one molecule of glucose is converted to
enter the mitochondrial electron transport two molecule of lactate, the net yield of 2
chain to complete oxidation molecules of ATP

• Each NADH provides 3 ATPs • 4 molecules of ATP are synthesised by

the 2 substrate level phosphorylation
• Since there are 2 NADH this reaction (steps 7 and 10)
provides 3x2= 6 ATPs
• But 2 ATP molecules are used in the steps
The net gain of energy from 1 and 3 the net yield is only 2 ATP. (it
glycolysis pathways is 8 ATPS returns NADH back to NAD+)

Glycolysis metabolic disorders

• Glycolytic mutations are generally rare due to importance of the metabolic pathway.

• Mutations of most of the enzymes in the glycolytic pathway have been described in

association with hemolytic anaemia.

1. Hereditary red cell enzymopathies: Pyruvate kinase deficiency

enzyme disorders is deficiency of pyruvate kinase (PK).

2. Chronic haemolytic anemia (CHA): Hexokinase defficiency

Hexokinase (HK) deficiency are very similar to those of pyruvate

kinase (PK) deficiency

This defect will also result in anaemia

 The Central Role of the Citric Acid Cycle
Three processes play central roles in aerobic metabolism: Play an
essential role in the production of energy by a cell
1. The citric acid cycle
2. Electron transport reactions
3. Oxidative phosphorylation

Metabolism consists of,

1. Catabolism: the oxidative breakdown of nutrients
2. Anabolism: the reductive synthesis of biomolecules
The Citric Acid Cycle is amphibolic: that is, it plays a role in both catabolism and

anabolism. It is the central metabolic pathway

-After glycolysis, 90% of energy in glucose is still unused, locked in pyruvate

-Oxygen is required to get the remaining energy

-During krebs cycle, pyruvate is broken down into CO2 through many steps.

Citric acid cycle: is a series of chemical reactions in mitochondria used by all

aerobic organisms to release stored energy through the oxidation of acetyl CoA

derived from carbohydrates. fats, and proteins into carbon dioxide and chemical
energy in the form of adenosine triphosphate (ATP).

Names:
1. Krebs Cycle
2. Citric Acid Cycle
3. Tricarboxylic Acid Cycle (TCA)

Where does it take place?

In eukaryotes, the enzymes of TCA are located in the mitochondrial matrix.

Kerbs Cycle: Preparatory Step conversation of

Pyruvate to Acetyl CoA
Pyruvic acid from glycolysis is converted to acetyl coenzyme A ( A-CoA) in 3
main steps;
1. Decarboxylation
1 carbon is removed from pyruvaic acid, 3C & 2C molecule
The lost carbon forms carbon dioxide (CO2); exhaled
2. Oxidation
A Hydrogen atom are removed from pyruvaic acid (‘oxidation’) and picked
up by NAD
NAD is reduced to NADH + H
3. Formation of acetyl CoA
The resulting acetic acid is combined with coenzyme A, a sulfur-
containing coenzyme, to form acetyl CoA (A-CoA)
Pyruvate dehydrogenase complex is responsible for the conversion of pyruvate to
CO2 and the acetyl portion of acetyl COA

Pyruvate dehydrogenase complex is 5 Enzymes in complex

1. Pyruvate dehydrogenase
2. Dihydrolipoyl transacetylase
3. Dihydrolipoyl dehydrogenase
4. Pyruvate dehydrogenase kinase
5. Pyruvate dehydrogenase phosphates

Pyruvate conversion into acetyl CoA requires a few cofactors in addition to the enzymes that
make up the complex; including thiamine pyrophosphate (TPP), lipoamide, FAD and
NAD+.
Summary
• The two-carbon unit needed at the start of the citric acid cycle is obtained
by converting pyruvate to acetyl-COA
• This conversion requires the three primary enzymes of the pyruvate
dehydogenase complex, as well as, the cofactors TPP, FAD+, NAD+,
and lipoic acid
• The overall reaction
Features of TCA Cycle
• For every glucose molecules two pyruvate molecules are produced by glycolysis
• Transferred into mitochondria where decarboxylation leads to formation of two acetyl-CoA and two C02
• Once it is produced, from whatever precursor, it is degraded viaTCA a cycle
Reactions of the Citric Acid Cycle
IN GENERAL:
TCA cycle is an eight-step cycle in which each acetic acid is decarboxylated and oxidized, generating:

- Three molecules of NADH + H+ (ox/red)

- One molecule of FADH2 (ox/red)

- Two molecules of CO (decarboxylation)

- One molecule of ATP (substrate level phosphorylation)

• In step 1, there is a condensation of acetyl- Co with oxaloacetate to form citrate

• Reaction is catalyzed by citrate synthase.
• Citrate synthase is an allosteric enzyme that is
• inhibited by NADH, ATP, and succinyl-CoA

• In step 2, citrate is isomerized to isocitrate

• This step is accomplished by a dehydration step followed by a
hydration step.
• The reaction is catalyzed by aconitase
• In step 3, Formation of a-ketoglutarate and Co2 by oxidative
decarboxylation reaction.
• There is an oxidation of isocitrate followed by decarboxylation to
form B-ketoglutarate and CO2
• The reaction is catalyzed by isocitrate dehydrogenase.
• Isocitrate dehydrogenase is an allosteric enzyme, which is inhibited
by ATP and NADH, and activated by ADP and NAD

• In step 4, formation succinyl -CoA by oxidative decarboxylation

reaction.
• This reaction is catalyzed by the a-ketoglutarate dehydrogenase
complex, which is, like pyruvate dehydrogenase, a multienzyme
complex and requires:
-coenzyme A,
-thiamine pyrophosphate TPP
-lipoic acid.
-FAD, and NAD+

• In step 5 the thioester bond of succinyl-COA is hydrolyzed to

form coenzyme A and succinate
• The two CH2-COO- groups of succinate are equivalent
• This is the first energy-yielding step of the cycle (substrate-level
phosphorylation)
• The overall reaction is slightly exergonic
• This reaction is catalyzed by the succinyl-CoA synthestase.
• In step 6, oxidation of succinate to fumarate
• Succinate dehydrogenase is a FAD linked enzyme bound to
the inner mitochondria membrane
• require Fe as cotactor

• In Step 7, the hydrotion of fumarate to L-maltate occurs

• This reaction is catalyzed by the Fumarase.

• In step 8, malate is oxidized to Oxaloacetate

• This reaction is catalyzed by the malate dehydrogenase.
 Summary
• Acetyl CoA links glycolysis and pyruvate oxidation with the citric acid cycle.
• In the citric acid cycle, the two carbons that were originally the acetyl group
of acetyl CoA are released as carbon dioxide, one of the major products of
cellular respiration, through a series of enzymatic reactions.
• The release of the two carbon dioxide molecules are coupled with:
◦ the production of (3)NADH from the reduction of NAD+
◦ the production of FADH2 from the reduction of FAD
◦ the production of one GTP/ATP

Metabolic regulation of
the Citric Acid Cycle

There are 3 control point within There is 1 control point outside

the cycle the cycle
Citrate synthase “step 1”
• inhibited by; ATP, NADH, succinyl CoA
• Feedback inhibition by; Citrate

Pyruvate dehydrogenase
• inhibited by; ATP, NADH
• Feedback inhibition by; Acetyl-CoA
Isocitrate dehydrogenase “step 3”
• inhibited by; ATP, NADH
• Activated by; ADP , NAD+

Ketoglutarate dehydrogenase complex “step 4”

• inhibited by; ATP, NADH, succinyl CoA
• Activated by; ADP , NAD+
• Pyruvate is oxidatively decarboxylated to acety| CoA, which enters into the
citric acid cycle. Complete oxidation of glucose through glycolysis plus
citric acid cycle will yield a net 38 ATPs.

• Energy yield (number of ATP generated) per molecule of glucose when it

completely oxidized through glycolysis pathway plus citric acid cycle. under
aerobic conditions

• in eukaryotic cells cytosolic NADH, loses an electron and enter the

respiratory chain as FADH2 therefore the ATP yield from these 2 NADH is
2 not 3 this will results in total 36 ATP instead of 38.

The Citric Acid Cycle in

Anabolism Catabolism
• The catabolism of carbohydrates,
• The citric acid cycle is the source of starting Ipids, and proteins all feed into the
materials for the biosynthesis of other compounds citric acid cycle at one or more
• If a component of the citric acid cycle is taken out points
for biosynthesis, it must be replaced • In catabolic pathway, nutrients, many
• A reaction that replenishes a citric acid cycle of which are macromolecules, are
intermediate is called an anaplerotic reaction broken down to smaller molecules.
• Small molecules are processed
further, and the end products of
catabolism frequently enter the CAC
 The link to oxygen
• The citric acid cycle is considered part of the aerobic metabolic process because of its link to
the electron transport chain and oxidative phosphorylation

• NADH and FADH2, two important cofactors generated by the citric acid cycle, ultimately
pass their electrons to oxygen

Metabolism of Fructose
• The major source of fructose is the disaccharides sucrose: (glucose+fructose).

• The entry of fructose into cells is not insulin-dependent (unlike glucose), and in contrast to glucose,
fructose does not promote the secretion of insulin.

• Liver is the main site of fructose metabolism:

◦ Fructose breakdown
◦ Fructose formation

Reaction of Fructose Breakdown (Liver)

1. Fructose is phosphorylated to F-1-P by fructokinase in the liver
2. F-1-P is cleaved by liver aldolase (aldolase-B) to dihydroxy acetone phosphate (DHAP) and
glyceraldehyde.
3. Glyceraldehyde is phosphorylated by glyceraldehyde kinase to glyceraldehyde-3-phosphate

- According to the metabolic status of the tissue>>

these two triose phosphate (DHAP and GA-3-P) enter the glycotic pathway. gluconeogenesis and
glycogenesis.

Reaction of Fructose Breakdown (extrahepatic tissue)

• In extrahepatic tissue, especially the adipose and muscle tissues, fructose can be phosphorylated by
hexokinase to F-6-P.

• However, this is a slow reaction occurs in the presence of high concentration of fructose because when
fructose is present with glucose, its phosphorylation is inhibited by glucose.

Disorders of fructose metabolism

1. Essential fructosuria, caused by a deficiency of the enzyme Fructokinase.
2. Hereditary fructose intolerance = fructosemia, it is due to deficiency of the enzyme Aldolase-B.
 Metabolism of Mannose
(Conversion of mannose to fructose-6- phosphate)

1. The C-2 epimer of glucose is an important component of glycoproteins.

2. Hexokinases phosphorylate mannose, producing mannose-6- phosphate
3. Mannose-6-phosphate is Isomerised to F-6-P by phosphomannose
isomerase
 Sorbitol (polyol) pathway
(Conversation of glucose to fructose)

• Sorbitol is a sugar alcohol (polyol) commonly used as a sweetener in so-called sugar-free sweets and
chewing gum, diet and diabetic foods.

• Cells use glucose for energy. This normally occurs by phosphorylation via the enzyme hexokinase

>> However, if large amounts of glucose are present (as in diabetes mellitus), hexokinase becomes
saturated and the excess glucose enters the sorbitol (polyol) pathway.

• Sorbitol pathway is a two-step metabolic pathway in which glucose is reduced to sorbitol, which is
then converted to fructose
>> this pathway provides a way for the body to produce fructose from glucose without using ATP.

Sorbitol (polyol) pathway:

1. Aldose reductase reduce glucose to sorbitol.
◦ this enzyme is found in many tissues as lens, retina, peripheral nerve cells, kidney, placenta,
RBC, ovaries and liver. Reduction

2. In the cells of liver, ovaries and sperm cells, there is a second enzymes, sorbitol dehydrogenase
that can oxidize the sorbitol to produce fructose

◦ The pathway from sorbitol to fructose in the liver provides a mechanism by which dietary sorbitol
is converted into fructose which can enter glycolysis
 Galactose metabolism
• The major dietary source of galactose is lactose (glucose+galactose) obtained from milk and milk
product

• Some galactose can also be obtained by lysosomal degradation of complex carbohydrates such as,
glycoproteins and glycolipids, which are important membrane components.

• Like fructose, the entry of galactose into cells is not insulin-dependent

• Galactose is readily converted in the liver to glucose.

Reactions of the pathway

1. The first reaction in galactose metabolism in the liver is phosphorylation of galactose to galactose
phosphate by the enzyme galactokinase, using ATP as [phosphate donor]

2. Galactose-1- phosphate reacts with UDP-glucose to form: [UDP-galactose and glucose-1-

phosphate]. The reaction is catalyzed by galactose-1-phosphate uridyl transferase.

3. The conversion of UDP-galactose to UDP-glucose catalysed by an UDP-galactose-4- epimerase.

Epimerization involves an oxidation and reduction at carbon 4 with NAD+ as a coenzyme

4. Finally, glucose released from UDP-glucose after conversion to G-1-P via formation of glycogen by
glyogenesis is followed by glycogenolysis

Disorders of galactose metabolism

• Galactosemia Three defects have been described due to deficiency of:
◦ Galactokinase
◦ Galactose-1-phosphate uridyl transferase
◦ UDP-galactose-4-epimerase

• The commonest and most serve enzymatic defect is due to Galactose-1-phosphate uridyl
transferase

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