Carbohydrate Metabolism

Dietary Carbohydrates: mainly include sucrose (table sugar), lactose (in milk) and
starch, which is present in potatoes, rice, corn and wheat. Also, glucose and
fructose are present in fruits. Cellulose is present in the cell wall of some plants
but is not digested by humans as it contains β-glucosidic linkage not hydrolyzed
by human amylase. It increases the bulk of stools, thus preventing constipation.

Absorption of monosaccharides:

- Different monosaccharides, ingested in diet and resulting from the process of

digestion, can be absorbed through three main mechanisms:
1. Simple (Passive) diffusion:
- It depends on the concentration gradient of sugars between the intestinal lumen
and mucosal cells.
- Fructose and pentose are absorbed by this mechanism.

2. Facilitated transport (GLUT-5):

It requires a transporter. It is the main transporter for fructose. Also, galactose and
glucose can be absorbed by this transporter.
3. Cotransport (Active Transport):

- This is an active mechanism which needs energy derived from the hydrolysis of
ATP, as well as a transporter.
- By this process, glucose and galactose are actively transported against their
concentration gradients.
- The brush border of enterocytes contains a sodium glucose transporter (SGLT-
1), which binds both sodium ions and glucose at separate sites and transport
both through the brush border of intestinal cells and both are released into the
cytosol allowing the carrier to bind more sodium and glucose.At the basal
border, all sugars are transported by facilitative transport (GLUT-2).

Glucose Uptake by Different Tissues:

- Glucose is transported by different transporters which may facilitative or sodium
dependent transporters.(GLUT-1,GLUT-2,GLUT-3,GLUT-4 ,GLUT-5 ,SGLT-1
and SGLT-2)
- GLUT-2 is found in liver, kidney and B-cells of pancreatic islets. It is
characterized by rapid uptake and release of glucose.
-GLUT-4 (insulin- dependent) is present in heart, skeletal muscles and adipose
tissue where insulin produces its translocation to the outer cell membrane surface.

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 Fate of Absorbed Sugars:
A- Glucose:It is transported to the liver then by systemic blood to different
tissues. The main pathways for glucose utilization are as follows:
1- Oxidation for production of energy.
2- Provides other compounds as follows:
- Carbohydrates i.e. fructose, galactose, pentoses, amino sugars and uronic
acids.
- Glycerol 3-phosphate for synthesis of triacylglycerol and phospholipids.
- Active acetate for synthesis of cholesterol, steroids and fatty acids.
- Nonessential amino acids.

3- Storage
Glucose is stored as glycogen in liver and muscles and as triacylglycerol in
adipose tissue.
N.B.Glucose can be excreted in urine if its plasma level exceeded the renal
threshold (180 mg/dl).

Oxidation of glucose
The pathways for oxidation of glucose include:
I-Glycolysis:
It is the main pathway for glucose oxidation. It is mainly concerned with energy
production.
II- Other Pathways for Oxidation:are mainly for synthesis of other glucose
derivatives and not for energy production.
1- Hexose monophosphate pathway (HMP): For production of pentoses and
NADPH.
2- Uronic acid pathway: For production of uronic acids.

Glycolysis
Definition:
Glycolysis is a sequence of enzymatic reactions in which one molecule of glucose
is converted into:
- Two molecules of pyruvate in the presence of oxygen OR
- Two molecules of lactate in the absence of oxygen.

Site: cytosol of all cells.

Steps of Glycolysis can be divided into two phases:

- Energy utilization phase (I): This phase consumes 2 ATPs.

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- Energy recovery phase (II): This phase produces 4 ATPs at the substrate level
(aerobically & anaerobically) and 5 ATP under aerobic state only.
Phase –I

Mg2+

Mg2+

Phosphotriose
Isomerase

- In this phase 1 molecule of glucose (C6) is converted to 2 molecules of

glyceraldehyde 3-phosphate (C3) as follows:
ATP ATP
Glucose (C6) 2 Glyceraldehyde -3 -P (C3)
- These steps actually require energy, in the form of two ATPs per glucose
molecule.

Phase –II:
- In this phase the 2 molecules of glyceraldehyde 3-P are converted to 2
molecules of pyruvate (aerobic) or lactate (anaerobic):

2 ATP
2Pyruvic acid
(2)Glyceraldehyde-3 P (C3)
2 NADH, 2H+

NAD+

2 Lactic acid
3
 2
2

2
2
2
Mg2+
2
2

2
Mg2+
2
2
Mg2+
2
2 (Aerobic Oxidation)

2 NADH, 2H+
Lactate dehydrogenase
2 NAD+
2 Lactate (Anaerobic Oxidation)

The redox state of the tissue determines which of two pathways is followed:
a- Under aerobic conditions: Pyruvate is transported into mitochondria and after
conversion to acetyl-CoA is oxidized by the citric acid cycle.

b- Under anaerobic conditions: The reoxidation of NADH is catalyzed by lactate

dehydrogenase (LDH). The reoxidation 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 i.e.in red cells (no mitochondria) and in muscles during severe
muscular exercise.
(See the diagram below)

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 Glycolysis under Aerobic and Anaerobic States

Aerobic state Aerobic state

Glucose Pyruvate Active acetate

NAD+ NADH,H+
ETC

H2O [O]

TCA cycle
Anaerobic state Anaerobic state
Glucose Pyruvate Lactate
LDH
NAD+ NADH,H+ NADH,H+ NAD+

Importance of Glycolysis
I. Energy production
A- Under aerobic state, the net gain of chemical energy is 7 ATP molecules
in phase I and II as follows:
1- Two ATP molecules are utilized by glucokinase (or hexokinase) and
phosphofructokinase-1.
2- Four ATP molecules are produced by substrate level phosphorylation by
phosphoglycerate kinase and pyruvate kinase.
3- Five ATP molecules from oxidation of 2 NADH through ETC. These two
NADH are produced by glyceraldehydes 3-phosphate dehydrogenase.
B- Under anaerobic state, the net gain of chemical energy is 2 ATP
molecules only:
NADH produced is not oxidized by ETC (4 ATP produced – 2 ATP utilized = 2
ATP).

N.B. Glycolysis liberates only a small part of energy from glucose, however:

a. It is very important during severe muscular exercise, where oxygen supply is

often insufficient to meet the demands of aerobic metabolism.
b. It provides all energy required by the RBCs. (due to lack of mitochondria).

II. In Erythrocytes
Glycolysis is the only source of ATP by substrate level oxidative-phosphorylation
because it lacks a mitochondria.

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III. Importance of Glycolytic Intermediates
1. Pyruvate generated by glycolysis has several fates i.e. conversion to active
acetate, oxaloacetate, alanine and lactate.
2. 3-Phosphoglycerate can be converted into serine amino acid.
3. Dihydroxyacetone-phosphate can be converted into glycerol 3-phosphate
which is used in triacylglycerol and phospholipid synthesis.

Clinical applications of Glycolysis

Glyceraldehyde-3-phosphate dehydrogenase: it is inhibited by arsenite and

iodoacetate which block the SH group present in the active site of the enzyme.
Enolase: is inhibited by fluoride as it irreversibly binds to Mg2+ which is
present at the active site of the enzyme. So, fluoride is added to blood samples
before glucose estimation to prevent glycolysis.
Pyruvate Kinase: Its deficiency causes hemolytic anemia.

Regulation of Glycolysis (Allosteric and Hormonal Regulation)

It can be noted that all reactions of glycolysis are reversible except those
catalyzed by:
1. Glucokinase (GK) or hexokinase (HK).
2. Phosphofructokinase-1 (PFK-1)
3. Pyruvate kinase (PK)

Glycolysis is regulated by factors which control the activity of the key enzymes
which catalyze the 3 irreversible reactions.

1- Glucokinase (GK or hexokinase D) and Hexokinase (HK):

Hexokinase Glucokinase
(Hexokinase D)

Site Extrahepatic Liver and pancreatic β cells

Substrate Glucose mainly and Glucose mainly and

other hexoses other hexoses

Affinity to glucose Has a low Km i.e. high affinity Has a high Km i.e. low affinity
for glucose and low Vmax. for glucose and high Vmax.

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 Effect of :
-Insulin No Effect Inducer
-Glucagon No Effect Repressor

-Fasting No Effect Decreases the activity

No Effect Increases the activity
-Feeding of glucose

2- Phosphofructokinase-1 (PFK-1):
- PFK1 is the most important control site in the mammalian glycolytic pathway
because it is a committed step.
- PFK1 isallosterically inhibited by: High levels of ATP.
- PFK1 isallosterically activated by: AMP.
Regulation of Glycolytic Key Enzymes
3- Pyruvate Kinase (PK):
- PK is allosterically inhibited by ATP (a parallel
Glucose regulation with PFK-1).
- PK is allosterically activated by PEP and fructose 1,6-bisphosphate.
HK GK
Insulin
III-Effect of Fatty Acid Oxidation on Glycolysis:
- Fatty acid oxidation decreases the Glucose
rate of6-P
glycolysis by:
- Fatty acid oxidation produces ATP which inhibits PFK-1 and PK +
ATP oxidation produces active acetate which forms citrate. The later
- Fatty acid
Fructose 6-P
produces inhibition of PFK-1. GK
_
PFK-1
PFK-1 PK
AMP +

Fructose 1,6-BP
_

Glyceraldehyde 3-P
Glucagon
(Liver)
2-PEP

_ PK
ATP

Pyruvate

II- Hormonal Regulation:

Insulin/glucagon ratio is the main hormonal regulator of glucose utilization, it

increases during glucose feeding and decreases during fasting or glucose
deprivation.

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 a- Glucagon:
It is secreted in case of carbohydrate deficiency or in response to low blood
glucose level (hypoglycemia). It affects liver cells mainly (inhibitor of
glycolysis) as follows:

1. It acts as a repressor of glycolytic key enzymes (GK, PFK-1 and PK).

2. It binds to specific cell membrane receptors → activation of specific G proteins→
activation of the adenylyl cyclase → ↑cAMP→ activates the protein kinase A→
inactivation of PK .

b- Insulin:
- It is secreted after feeding of carbohydrates or in response to high blood glucose
level (hyperglycemia).
1. It acts as inducer for synthesis of the glycolytic key enzymes (GK, PFK-1 and PK).
2. It activates phosphodiesterase (↓cAMP → inhibition of protein kinase A).
It activates protein phosphatase → activation of PK.

Effect of Glucagon and Insulin on Enzymes that Regulate

Glycolysis Through cAMP
Glucagon Insulin
+ +
Adenylyl cyclase Phosphodiesterase
cAMP
ATP AMP
PPi H2O

Inactive protein kinase A Active protein kinase A

ATP ADP
Mg2+ PK- b (inactive)
PK- a (active)

Pi H2O
P
Protein
phosphatase
Activation of Glycolysis - Inhibition of Glycolysis
+ -
Insulin cAMP

III-Effect of Fatty Acid Oxidation on Glycolysis:

- Fatty acid oxidation decreases the rate of glycolysis. This is an important
mechanism to minimize oxidation of glucose by different tissues during fasting
or carbohydrate deprivation, which allows more glucose for the brain.
- Fatty acid oxidation produces ATP which inhibits PFK-1 and pyruvate kinase.

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 - Fatty acid oxidation produces active acetate which forms citrate. The later
produces inhibition of PFK-1.

Reversal of Glycolysis (Gluconeogenesis)

During fasting and low dietary carbohydrates, glycolysis is reversed for synthesis
of glucose from non-carbohydrate compounds.

Conversion of Pyruvate to Active Acetate and Oxaloacetate

A- Conversion of Pyruvate to Active Acetate
- Pyruvate formed at the end of glycolysis is transported into the mitochondria by a
specific pyruvate transporter, where it is converted by a process of oxidative
decarboxylation into acetyl-CoA (active acetate).

- This step is catalyzed by pyruvate dehydrogenasecomplex (PDH) which requires

five coenzymes: thiamine pyrophosphate (TPP), lipoate, CoA, FAD and NAD+.
- PDH is only active in aerobic condition as NADH produces inhibition of the
enzyme.
- In case of complete oxidation of glucose, two molecules of pyruvate form two
molecules of active acetate and two NADH. Two NADH will produce 5 ATP by
oxidation through ETC.

ADP
NAD+
Insulin NADH
Pyruvate
(in adipose tissue) Acetyl-CoA
CoA
Ca2+ ATP

+ + -
O O
Pyruvate Dehydrogenase Complex
CH3-C-COOH CH3-C~S-CoA
Pyruvate TPP Acetyl-CoA
NAD+ Lipoate NADH,H+
CoA-SH FAD CO2

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Regulation of Activity of PDH
I- PDH activity is regulated through allosteric mechanism as follows:
- It is inhibited by the product of the reaction (acetyl-CoA and NADH) and
ATP.
- It is activated by NAD+, Pyruvate and CoA (substrates of the reaction) and
ADP.
- It is activated by Ca2+

II- In adipose tissue: insulin increases the activity of PDH.

- During carbohydrate feeding: Insulin in adipose tissue increases the activity
of PDH. This directs pyruvate to active acetate and increases the rate of
glucose utilization for fatty acid synthesis (lipogenesis) and fat
storage.Accordingly the activity of the enzyme increases by feeding of
carbohydrate and decreases during fasting.

- During fasting or carbohydrate deprivation: Oxidation of fatty acids and

ketone bodies is increased which produces active acetate and ATP → decrease
the activity of PDH → decreased conversion of pyruvate to active acetate
(↓glucose utilization).

III-In muscles: During muscular exercise, Ca2+ is released from its storage
sites → activation of PDH →increased conversion of pyruvate to active
acetate (↑glucose utilization), which is important to supply more ATP for
contracting muscles.

N.B.
- Deficiency of PDH or thiamine (vitamin B1) results in an inability to convert
pyruvate to acetyl-CoA, and accumulation of pyruvate and its conversion to lactate
which leads to lactic acidosis.
- The condition is mainly fatal. It produces brain damage as the brain is dependent
mainly on glucose for energy production and is particularly sensitive to acidosis.

II- Conversion of Pyruvate to Oxaloacetate

This reaction is catalyzed by the mitochondrial enzyme pyruvate carboxylase
(PC).
O ATP Biotin
CO2 ADP + Pi
2+
Mg O = C - COOH
CH3-C-COOH
CH2 - COOH
Pyruvate Pyruvate carboxylase Oxaloacetate

Oxaloacetate has an important catalytic function in oxidation of active acetate by

citric acid cycle.

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I- Allosteric Regulation
Active acetate: It acts as an allosteric activator of pyruvate carboxylase, this is
important becauseoxaloacetate has an important catalytic function in oxidation of
active acetate by citric acid cycle.

II- Hormonal Regulation

1- Antiinsulin hormones
- Conversion of pyruvate to oxaloacetate is an important step during
gluconeogenesis.
- So, the activity of pyruvate carboxylase is increased by anti-insulin
hormones (mainly glucagon, epinephrine and glucocorticoids), which act as
inducers for the enzyme.
2-Insulin
It acts as a repressor for pyruvate carboxylase and decreases its synthesis; this
effect does not decrease the rate of oxidation of active acetate by citric acid cycle
because the enzyme is allosterically activated by active acetate.

Action and Regulation of Activity of Pyruvate Carboxylase

O ATP Biotin
CO2 ADP + Pi
2+
Mg O = C - COOH
CH3-C-COOH
Pyruvate carboxylase CH2 - COOH
Pyruvate Oxaloacetate
- +
+
Repressor Inducers Allosteric activator
Insulin Anti-insulin Active acetate
(Glucagon, epinephrine & glucocorticoids)

Energy Yield from Glucose Oxidation

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Oxidation of one mole of glucose yields up to 32 moles of ATP under aerobic
conditions, but 2 moles only under anaerobic condition.

Complete Oxidation of glucose Summary for

Glucose
(One molecule)

Glycolysis

2 Pyruvate + 7 ATPs

PDH
CO22

2 Active acetate + 5 ATPs

7ATPs
Oxidation by citric acid cycle

4 CO2+ 20ATPs

Net result = 6 CO2 + 32 ATPs

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 Hexose Monophosphate Pathway (HMP)
Pentose Phosphate Pathway (PPP)

Definition:
The pentose phosphate pathway is an alternative route for glucose oxidation. It has
two major functions, formation of NADPH and ribose 5-phosphate.

Site:
- The enzymes of the pentose phosphate pathway are cytosolic.
- HMP is active in certain tissues e.g. liver, thyroid, adrenal cortex, adipose tissue,
gonads, retina, lactating mammary gland and RBCs.

Steps:
The sequence of reactions of the pathway can be divided into two phases:
1-Oxidative irreversible phase: In this phase, glucose 6-phosphate undergoes
dehydrogenation and decarboxylation to yield a pentose i.e.ribulose 5-phosphate.

2-Nonoxidative reversible phase: In this phase, six molecules of ribulose 5-

phosphate are converted back to five molecules of glucose 6-phosphate by a series
of reactions involving mainly two enzymes: transketolase and transaldolase

6 Glucose-6-P Oxidative phase 6 Ribulose-6-P

(C36) (C30)
12 NADP+ 12 NADPH, H+ 6 CO2

Nonoxidative phase
5 Glucose-6-P
(C30)

N.B. Glucose-6-P dehydrogenase (G6PD) is the key enzyme of the oxidative

phase.

Importance of HMP pathway

1- It provides ribose 5-phosphate which is required for synthesis of nucleotides
and nucleic acids. In tissues (e.g. muscles) where the oxidative HMP is not
active due to the deficiency of the enzymes of the oxidative phase and in cases of
favism (deficiency of G6PD), pentoses are formed by the reversal of the non-
oxidative phase.
2- It is the main source of NADPH, which is required for the reaction of many
reductases, hydroxylases and NADPH oxidase. which catalyze several important
biochemical reactions, e.g.:

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 a. Fatty acid synthesis (in liver, adipose tissue and lactating memory gland).
b. Steroid synthesis (in adrenal cortex, testis, ovaries and placenta).
c. Important for vision (in the eye)
d. Importance in RBCs
Glutathione reductase and glutathione peroxidase are important for removal of
hydrogen peroxide (H2O2). H2O2 produced through certain biochemical
reactions is a powerful oxidant and produces damage of cellular DNA,
proteins and phospholipids of the cell membrane

Glutathione peroxidase
H2O2 2 H2O
Se
2 G – SH G-S – S-G

NADP+ NADPH,H+
Glutathione reductase
(FAD)

Favism
Prevalence: Favism is the most common human enzymopathy.
Cause: Favism is due to genetic deficiency of glucose-6-phosphate dehydrogenase
(G6PD).
Mechanism:
1. Red cells are liable for oxidative damage by H2O2 due to their role in oxygen
transport. H2O2 produces lipid peroxidation, which increases the cell membrane
fragility.
2. The red cell capacity to protect itself from oxidative damage is markedly
decreased due to decreased concentration of NADPH which is required by
glutathione reductase for the regeneration of reduced glutathione (GSH)for
removal of H2O2 (by glutathione peroxidase).
3. Exposure of red cells to oxidizing agents produces lysis of red cells and
development of hemolytic anemia and jaundice.
4. Administration of certain drugs (premaquine, aspirin or sulfonamides) or eating
Fava beans (containing oxidants), stimulate the production of H2O2 and produce
lysis of the fragile red cells.
Treatment: The only treatment is to avoid the above factors and by blood transfusion
during the attack of hemolysis.

Regulation of HMP pathway:

I-Allosteric regulation

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NADPH is a strong inhibitor of glucose 6-phosphate dehydrogenase.
II-Hormonal regulation
The synthesis of G6PD isinduced by the increased insulin/glucagon ratio after
a high carbohydrate meal.

URONIC ACID PATHWAY

Definition:
Uronic acid pathway is also an alternative oxidative pathway for glucose. In
humans, it catalyzes the conversion of glucose to glucuronic acid, and pentoses.
Site:
Uronic acid pathway is a cytosolic pathway that occurs inthe liver.
Importance of Uronic acid pathway
The main function is the formation of UDP-glucuronate which is utilized in the
following pathways:
1) Glycosaminoglycans (GAGs) synthesis.
2) Conjugation reactions with many compounds to increase their water solubility
such as: all steroid hormones and their metabolites, bilirubin and certain
detoxification reactions of xenobiotics such as phenols.

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