University of Duhok

College of Medicine
Metabolism module

Session 3
Lecture 1+2
Carbohydrate metabolism part 2

Dr. HIVI M. MAHMOUD

M.B.,Ch.B., M.Sc., PhD. (Clinical Biochemistry)
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 Objectives

1. Definition of the HMP pathway

2. Major differences between EM and HMP pathway
3. Reactions of HMP pathway
4. Physiological significance of the HMP pathway
5. Clinical significance of the HMP pathway, G6PD
6. Role of TCA and how is regulated
7. How and why glucose is formed from non-
carbohydrate sources (Gluconeogenesis)

8. Galactose metabolism and biochemical and clinical

basis of galactosaemia
9. Fructose metabolism.

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Definition of the shunt pathway

It is also known as:

pentose phosphate pathway or hexose monophosphate shunt

instead of glucose going through glycolytic pathway, it is shunted

through this pathway.

Defined as an alternative pathway for oxidation of glucose

About 10% of glucose per day are entering in HMP shunt pathway

It occurs in the cytosol of the cell,

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The major purpose of this pathway are generation of:

1-reduced nicotinamide adenine dinucleotide phosphate (NADPH)

production which used as biochemical reductant

2-pentose phosphate
which used for nucleotide synthesis

Three molecules of glucose -6-posphate enter the cycle, producing

3 molecules of CO2 and
3 molecules of 5 carbon residues which give
2 molecules of glucose 6 phosphate and
1 molecule of glyceraldehydes -3-phosphate

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Major differences between EM and HMP pathway

EM pathway
occurs in all tissues
oxidation by dehydrogenation but NAD is hydrogen acceptor
ATP is required and ATP is produced
CO2 is never formed

HMP pathway
occurs in certain special tissues for special function
oxidation by dehydrogenation but NADP is hydrogen acceptor
ATP is required and ATP is not produced
CO2 is produced

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 Reactions of Pentose Phosphate Pathway
Stage 1 oxidative reactions (irreversible)

Three reactions:

1-dehydrogenation of glucose 6-phosphate

2-hydration of phosphogluconolactone
3-formation of ribulose 5-phosphate (second dehydrogenation)

For each molecule of glucose 6-phosphate oxidization:

a-ribulose 5-phosphate.

b-CO2.

c-two molecules of NADPH 8

LO 3

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Stage 2 nonoxidative reactions (reversible)

These reversible reactions permit ribulose 5-phosphate

(produced by the oxidative portion of the pathway) to be
converted Either to

ribose 5-phosphate for nucleotide synthesis

or to

fructose 6-phosphate (intermediates of glycolysis)

or to

Glyceraldehyde 3-phosphate (intermediates of glycolysis)

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Physiological significance of the HMP pathway

the oxidative reactions of the pentose phosphate pathway is operating

in following organs:
a-liver

b-lactating mammary glands

c-adipose (which are active in the biosynthesis of fatty acids)

d-adrenal cortex (which is active in the NADPH-dependent synthesis

of steroids)
e-erythrocytes ( which require NADPH to keep glutathione reduced).

f-testes and ovaries

g-lens of eyes

the nonoxidative reactions of the pentose phosphate pathway operating

in
all cell types synthesizing nucleotides and nucleic acids.
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1-generation of reducing equivalents

NADPH enhance reductive biosynthesis of

fatty acid
cholesterol
steroids

2-free radical scavenging

NADPH enhance the regeneration of

reduced glutathione reductase which has a role in
removing the free radical (super oxide, hydrogen peroxide).

3-erythrocyte membrane integrity

NADPH is required by the red blood cells to

keep the glutathione in reduced state which
detoxify the free radical formed within the RBC

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4-prevention of methemoglobinemia

NADPH is required to keep

iron of Hb in reduced state (ferrous) and prevent
accumulation of methemoglobinemia (cannot carry oxygen)

5-lens of eye

NADPH is required for preserving the transparency of lens

maintenance of lens protein

6-macrophage bacteria activity

NADPH is required for production of

superoxide anion radical by macrophage to
kill bacteria.

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Clinical significance of the HMP pathway LO 5
Glucose 6-phosphate dehydrogenase deficiency

 Most common enzyme deficiency seen in clinical practice

 Inherited (X-linked) disease

 Hemolytic anemia which is caused by inability to detoxify oxidizing agents

diminished G6PD activity impairs the ability of the cell to form the NADPH
which is essential for the maintenance of the reduced glutathione pool,
resulting in decrease in the cellular detoxification of free radicals and
peroxides formed within the cell.

 Although G6PD deficiency occurs in all cells of the affected individual, it is

most severe in erythrocytes, where the pentose phosphate pathway provides
the only means of generating NADPH

hemolytic anemia which is characterized by

ANEMIA JAUNDICE BLACK URINE
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Precipitating factors in G6PD deficiency LO 5

Most individuals who have inherited one of the many G6PD mutations do
not show clinical manifestations (depend on the amount of deficiency of
enzyme activity)

some patients with G6PD deficiency develop hemolytic anemia if they

are:

Treated with an oxidant drug.

oAntibiotics (chloramphenicol)
oAntimalarials (primaquine)
oAntipyretics (acetanilid ) …….etc

Ingesting fava beans (Favism).

Severe infection.

The generation of free radicals by inflammation in macrophages, which

can diffuse into the red blood cells and cause oxidative damage. 16
 LO 6

Citric Acid Cycle

Tricarboxylic acid cycle

Krebs cycle
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 The aerobic processing of glucose starts with the complete oxidation
of glucose derivatives to CO2 and H2O.

 It occurs in mitochondria

 The Citric acid cycle is the final common central pathway for the
oxidation of fuel molecules –amino acids, fatty acids& CHO.

 Most fuel molecules enter the cycle as Acetyl CoA.

 Under aerobic conditions, the pyruvate generated from glucose is

oxidatively decarboxylated to Acetyl CoA.

 No known genetic defects that if present would be lethal.

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Biosynthetic roles of the citric acid cycle

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Regulation of TCA Cycle

The TCA cycle is controlled by the regulation of several enzyme

activities

The main drive for the cycle is

ATP/ADP:ratio and NADH/NAD+ ratio

The most important of these regulated enzymes are :

 Citrate synthase

 Isocitrate dehydrogenase

 Alpha ketoglutarate dehydrogenase complex

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Central nervous system and other glucose-dependent

tissues need glucose during fasting & starvation when the
glucose is absent from diet.

Initially this comes from the breakdown of liver glycogen

(glycogenolysis). However, the amount of glucose stored as
liver glycogen is only sufficient for 8-10 hours of fasting

If fasting lasts more than 8-10 hours the body has to

produce glucose by the process of
GLUCONEOGENESIS.
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• GLUCONEOGENSIS: LO 7

• The synthesis of glucose from noncabohydrate precursors is called GLUCONEOGENSIS.

 Liver is the major site of GLUCONEOGENSIS.

 Kidney: small percentage of GLUCONEOGENSIS .
 Brain: very little.
• A longer period of starvation, glucose must be formed from non-CHO sources such as
lactate, amino acids& glycerol).

MAIN GLUCONEOGENESIS SUBSTRATES:

 lactate is formed by the active skeletal muscle when the rate of glycolysis exceeds the rate of
oxidative metabolism,be reconverted to glucose by GLUCONEOGENSIS in the liver.

 amino acids are derived from proteins in the diet &, during starvation, from the breakdown
of proteins in the skeletal muscles.

 Glycerol is produce from the hydrolysis of triglycerides. 24

. LO 7

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Regulation of gluconeogenesis LO 7

 Gluconeogenesis occurs as part of the response to stress situations (e.g. fasting,

starvation, prolonged exercise) and is largely under hormonal control.

 The major control sites are PEPCK and Fructose 1,6-bisphosphatase.

 The activity of PEPCK is increased by glucagon and cortisol and decreased by

insulin.

 The activity of Fructose 1,6- bisphosphatase is also increased by glucagon and

decreased by insulin.

 Thus, the insulin/glucagon ratio plays a major role in determining the rate of
gluconeogenesis.

 In the absence of adequate levels of biologically effective insulin,

such as occurs in DIABETES, increased rates of gluconeogenesis
contribute significantly to the hyperglycaemia.
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Regulation of gluconeogenesis

1-GLUCAGON HORMONE stimulates gluconeogenesis by three

mechanisms.
a. Changes in allosteric effectors: Glucagon lowers the level of
fructose 2,6-bisphosphate, resulting in activation of fructose
1,6-bisphosphatase and inhibition of phosphofructokinase-1,
thus favoring gluconeogenesis over glycolysis.

b. Covalent modification of enzyme activity

c. Induction of enzyme synthesis: Glucagon increases the

transcription of the gene for PEP-carboxykinase.

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2-substrate availability: The availability of gluconeogenic
precursors, particularly glucogenic amino acids, significantly
influences the rate of hepatic glucose synthesis.

3-inhibition by AMP: Fructose 1,6-bisphosphatase is inhibited

by AMP—a compound that activates PFK.

4-activation by acetyl CoA: Activation of hepatic pyruvate

carboxylase by acetyl CoA occurs during fasting. As a result of
increased lipolysis in adipose tissue, the liver is flooded with
fatty acids.

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Galactose metabolism
 Dietary lactose is hydrolysed by the digestive enzyme lactase to release glucose
and galactose that are absorbed into the blood stream.

 Galactose is metabolised largely in the liver (some in kidney and gastrointestinal

tract)

 The epimerase reaction is reversible enabling galactose to be synthesized from

glucose via UDP-glucose.

 This is important during lactation when breast tissue is synthesizing large

amounts of lactose for milk production.

 There are a number of clinical conditions that affect galactose metabolism

including Lactose Intolerance and Galactosaemia.
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Galactosaemia
Unablity to utilize galactose obtained from the diet because of a lack of
the kinase or transferase enzyme.

The absence of the kinase is relatively rare (Duarte type) and is

characterized by accumulation of galactose in tissues.

The absence of the transferase is more common (classical type) and

more serious as both galactose and galactose 1-phosphate accumulate
in tissues.

Accumulation of galactose in tissues leads to its reduction to galactitol

(aldehyde group reduced to alcohol group) by the activity of the enzyme
aldose reductase:

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 Aldose reductase LO 8

 This reaction depletes some tissues of NADPH.

 In the eye the lens structure is damaged, (cross linking of lens

proteins by S-S- bond formation) causing cataracts. In addition, there
may be nonenzymatic glycosylation of the lens proteins because of
the high concentration of galactose and this may contribute to the
cataract formation.

 The accumulation of galactose and galactitol in the eye may lead to

raised intra-ocular pressure (glaucoma) which if untreated may cause
blindness.

 Accumulation of galactose 1-phosphate in tissues causes damage to

the liver, kidney and brain and may be related to the sequestration of
Pi making it unavailable for ATP synthesis. 32
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Fructose metabolism
 Dietary sucrose is hydrolysed by the digestive enzyme sucrase to release
glucose and fructose. These are absorbed into the blood stream.

 Fructose is metabolized largely in the liver by soluble enzymes that catalyze its
conversion to glyceraldehyde 3-phosphate (intermediate of glycolysis).

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THANK YOU
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