Carbohydrate Metabolism

Metabolism
of
Carbohydrates
Dr. Itepu, Victor E.
Mbbs, MSc
1
Outline
• Introduction
• Glycolysis
• Entry of other sugars into the glycolytic pathways
• Formation and fate of pyruvate
• Tricarboxylic acid (Krebs) cycle
• Gluconeogenesis
• Glycogenesis
• Glycogenolysis
• Pentose phosphate pathway
• Electron transport chain
• Convergence of metabolic pathways
2
Learning outcomes
At the end of the lectures, students should be able to:
• Describe the major and minor metabolic pathways of
carbohydrate metabolism.
• Describe how energy in the form of ATP is generated from the
various pathways.
• Demonstrate the knowledge of carbohydrate metabolism in
the detection and prevention of metabolic diseases including
diabetes mellitus.
• Discuss the role of carbohydrate in blood grouping.
3
Digestion of carbohydrates
• Dietary carbohydrates principally consist of the polysaccharides
and disaccharides.
• Liquid food materials like milk, soup, fruit juice escape digestion in
mouth as they are swallowed.
• Solid foodstuffs are masticated thoroughly before they are
swallowed.
• Cooking makes the digestion process easier.
• The digestion of carbohydrates occurs briefly in the mouth and
largely in the intestine.
• The hydrolysis of glycosidic bonds is carried out by a group of
enzymes called glycosidases.
4
Digestion in the mouth
• The digestion of carbohydrates begins in the mouth as the food
comes in contact with saliva during mastication.
• Saliva contains the enzyme salivary α-amylase (ptyalin) which
hydrolyzes α-1 → 4 glycosidic linkage in starch at random sites.
• Ptyalin requires Cl– ion and optimum pH 6.7 for activation.
• The products formed include α-limit dextrins, (containing about 8
glucose units with one or more α-1,6-glycosidic bonds) maltotriose
and maltose.
• The action of ptyalin stops in the stomach as a result of the pH of
3.
• Gastric juice does not contain any carbohydrate splitting enzymes.
• Hence, there is no action on carbohydrates in the stomach.
5
Digestion in the small intestine
• The pancreas secretes pancreatic juice into the duodenum.
• Pancreatic juice contains pancreatic α-amylase, which acts similar to
ptyalin.
• The acidic content of the stomach on entering the duodenum is
neutralized by bicarbonate produced by the pancreas.
• Pancreatic α–amylase also requires Cl– for activation.
• The enzyme hydrolyses α-1→4 glycosidic linkage situated well inside
polysaccharide molecule.
• The resultant products at this stage are disaccharides and
oligosaccharides.
6
Digestion in the small intestine cont’d
• The resultant disaccharides and oligosaccharides are digested at the
mucosal lining of the proximal jejunum.
• Here, oligosaccharidases (e.g. glucoamylase acting on amylose) and
disaccharidases (e.g. maltase, sucrase, lactase) act to convert these to
monosaccharides.
7
Absorption of carbohydrate
• Absorption of monosaccharides occurs in the duodenum and
proximal jejunum.
• Monosaccharides that are absorbed in decreasing order of relative
rate of absorption are galactose, glucose, fructose and mannose.
• However, glucose accounts for nearly 80% of the total
monosaccharides.
8
Mechanism of absorption
• Different sugars possess different mechanisms for their absorption.
• However, glucose and galactose are transported by the same
mechanism.
• Glucose is transported into the intestinal mucosal cells by the sodium
dependent glucose transporter (SGluT-1).
• Na+ concentration is higher in the intestinal lumen compared to the
intestinal mucosal cells.
• Na+ is carried by the carrier protein into the intestinal cells along its
concentration gradient and glucose is carried along with it.
• After delivering the glucose to the cells, Na+ is returned to the
intestinal lumen by the Na+-K+-ATPase pump.
• This return requires energy which is often supplied by ATP.
9
Mechanism of absorption cont’d
• Fructose is transported into the intestinal cells by facilitated diffusion
mediated by a carrier.
• It is relatively simple, is independent of energy and does not require
energy.
• Inside the cells, most of the fructose is converted to glucose, before
entering the circulation.
• Pentoses are absorbed by simple diffusion.
• Glucose present in the intestinal cells is transported into circulation
by means of facilitated diffusion using the Glucose Transporter Type-2
(GluT-2)
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Mechanism of absorption cont’d
• Unlike SGluT-1, GluT-2 is a uniport and as such, does not require
sodium to transport glucose.
• It transports glucose from the intestinal cells into capillaries using the
ping-pong mechanism.
• GluT-2 is also present in liver cells, beta cells of the pancreas and
renal cells.
• Another glucose carrier protein is GluT4, which is a major glucose
transporter in skeletal muscles and adipose tissue.
• Various other glucose carrier proteins are present in different cells of
the body, mediating the transport of glucose across cells.
• Some of these include GluT1 present in retina, RBC, kidney, etc,GluT3
present in neurons and brain, GluT5 in testis etc.
11
Carbohydrate metabolism
• Carbohydrate metabolism in mammalian cells can be classified into:
• Glycolysis: Oxidation of glucose to pyruvate (aerobic state) or lactate
(anaerobic state)
• Krebs (TCA) cycle: After oxidation of pyruvate to acetyl CoA, acetyl
CoA enters the Krebs cycle for the aim of production of ATP.
• Hexose monophosphate shunt: Enables cells to produce ribose-5-
phosphate and NADPH.
• Glycogenesis: Synthesis of glycogen from glucose, when glucose
levels are high
• Glycogenolysis: Degradation of glycogen to glucose when glucose in
short supply.
• Gluconeogenesis: Formation of glucose from non-carbohydrate
sources.
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Carbohydrate metabolism cont'd
• Glucose is the
major fuel of most
organisms.
• The major
pathways of
carbohydrate
metabolism either
begin or end with
glucose.
13
GLYCOLYSIS
• Glycolysis is the oxidation of glucose or glycogen to
produce pyruvate (under aerobic condition) and lactate
(under anaerobic condition).
• It is also called Embden Meyerhof pathway, being named
after Embden Meyerhof and Parnas who first described
it.
• Glycolysis occurs in almost all tissues and the reactions
occur in the cytoplasm.
• Red blood cells and cells of the nervous system derive
their energy mainly from glycolysis.
14
Glycolysis cont'd
• Anaerobic glycolysis forms the major source of energy for
muscles during strenuous exercise, when there is
insufficient oxygen.
• The glycolytic pathway provides carbon skeletons for
synthesis of non-essential amino acids as well as glycerol.
• Glycolysis consists of ten different reactions.
• However, the tenth reaction occurs under anaerobic
condition.
• Most of the reactions of the glycolytic pathway are
reversible.
• Consequently, they are also used for gluconeogenesis.
15
Reaction 1 of Glycolysis
• Glycolysis begins with glucose and in step 1, there is
phosphorylation of glucose to glucose-6-phosphate.
• The reaction involved the splitting of ATP to ADP.
• This reaction is irreversible and is catalyzed by
hexokinase, a key glycolytic enzyme, which has an iso-
enzyme glucokinase, that is only present in liver cells
unlike the former that is present in all cells.
• This reaction traps glucose in the cell.
• Hence, when glucose is phosphorylated, it must be
metabolized.
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Reaction 1 of Glycolysis cont'd
• 1 ATP is used up in this reaction for one molecule of glucose.
17
Step 2 of Glycolysis
• In step 2, there is isomerization of glucose-6-phosphate by
phosphohexose isomerase to produce fructose-6-phosphate.
• This reaction is reversible.
18
• In step 3, there is further phosphorylation of fructose-6-
phosphate to produce fructose-1,6-bisphosphate.
• The prefix bis- is used in place of di- because the
phosphorylation occurs at different positions.
• This reaction is irreversible and is catalyzed by the
allosteric, inducible and regulatory enzyme,
phosphofructokinase (PFK), which is a key glycolytic
enzyme.
• This is the rate limiting step of glycolysis.
19
Step 3 of Glycolysis cont'd
• ATP and citrate are the most important allosteric inhibitors of
PFK.
• AMP acts as an allosteric activator.
• In this reaction, another ATP molecule is used up.
20
• In step 4, a molecule of fructose-1,6-bisphosphate is
cleaved into 2 molecules of 3-carbon units, namely
glyceraldehyde-3-phosphate and dihydroxyacetone
phosphate (DHAP).
• The reaction is reversible and is catalyzed by aldolase.
• Dihydroxy acetone phosphate is isomerised to
glyceraldehyde-3-phosphate by the enzyme
phosphotriose isomerase.
• Thus, net result is that glucose is now cleaved into 2
molecules of glyceraldehyde-3-phosphate
21
22
• In step 5, there is dehydrogenation and
phosphorylation of glyceraldehyde-3-phosphate to
produce 1,3-bisphosphoglycerate, using a molecule of
NAD+.
• The reaction is reversible and is catalyzed by
glyceraldehyde-3-phosphate dehydrogenase.
• Two molecules of NADH are formed in this step.
• Under aerobic condition, each NADH can produce 2.5
ATPs via the mitochondrial electron transport chain,
making a total of 5 ATPs.
23
• Energy is derived in this step via oxidative phosphorylation in the
electron transport chain.
24
• Step 6 is a substrate level phosphorylation reaction.
• Here, energy is trapped directly from the substrate,
without going through the electron transport chain.
• A molecule of ATP is generated from each molecule of
1,3-bisphosphoglycerate, as it is being converted to 3-
phosphoglycerate.
• Reaction is reversible and catalyzed by
bisphosphoglycerate kinase.
• Therefore, 2 ATPs are formed from one molecule of
glucose at this step.
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• In step 7, 3-phospho glycerate is isomerized to 2-phospho
glycerate by the shifting of the phosphate group from 3rd to 2nd
carbon atom.
• The reaction is reversible and catalyzed by phosphogluco
mutase.
26
• In step 8, 2-phosphoglycerate is converted to
phosphoenol pyruvate (PEP).
• In this reaction, one molecule of water is removed.
• The reaction is reversible and catalyzed by the enzyme
enolase, which requires Mg²+.
• Hence, the enzyme can be inhibited by flouride, because
it will remove Mg²+, thereby halting glycolysis.
• This is the reason blood sample to be used for estimation
of blood glucose is collected in a flouride oxalate
container.
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• Step 9 is another substrate phosphorylation reaction of
glycolysis.
• Here, phosphoenol pyruvate (PEP) is dephosphorylated
to pyruvate.
• The reaction is irreversible and catalyzed by pyruvate
kinase, which is also a key glycolytic enzyme.
• One ATP is formed in this step from one molecule of PEP,
making a total of 2 ATPs from one molecule of glucose.
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• Step 10 reaction occur only in anaerobic conditions.
• Under aerobic condition, pyruvate enters the citric
acid/kreb cycle for complete oxidation.
• However, in step 10, pyruvate is reduced to lactate.
• The reaction is reversible and is catalyzed by lactate
dehydrogenase (LDH)
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Summary of glycolysis
33
Energetics of glycolysis
• Under aerobic
condition, 9 ATPs are
produced, both from
substrate level and
oxidative
phosphorylation but 2
ATPs are used up in
steps 1 and 3, resulting
in a net yield of 7 ATPs.
34
Energetics of glycolysis cont'd
• Under anaerobic
condition, 4
molecules of ATP are
synthesized by the 2
substrate level
phosphorylations
(steps 6 and 9).
• But 2 molecules of
ATP are used in the
steps 1 and 3, hence
the net yield is only
2 ATP.
35
Regulation of glycolysis
• Glycolysis is regulated by the three regulatory or key enzymes of
glycolysis, which are;
• Glucokinase/Hexokinase, step 1
• Phosphofructokinase, step 3
• Pyruvate kinase, step 9
36
Entry Of Other Sugars Into The Glycolytic Pathway
• Fructose
• Most of the fructose ingested is metabolized by the liver using
the fructose-1-phosphate pathway.
• The first step in the pathway is the phosphorylation of fructose to
fructose-1-phosphate by fructokinase.
• Fructose-1-phosphate then undergoes hydrolysis by fructose-1-
phosphate aldolase to form dihydroxyacetone phosphate (DHAP),
an intermediate in glycolysis and glyceraldehyde.
• DHAP is then isomerized to glyceraldehyde 3-phosphate by triose
phosphate isomerase to continue in the glycolytic pathway, while
glyceraldehyde is phosphorylated to glyceraldehyde-3-phosphate
by triose kinase.
37
Entry of Fructose into the glycolytic pathway
• Frucose can be
phosphorylated to
fructose-6-phosphate by
hexokinase.
• However, the affinity of
hexokinase for glucose is
20 times more than that
for fructose.
• Hence,most of the
fructose in adipose is
often metabolized in the
adipose tissue through
fructose-6-phosphate
38
Entry of Galactose into the glycolytic pathway
• Galactose is converted into glucose-6-phosphate, which enters
the glycolytic pathway, in four steps.
• The first reaction is the phosphorylation of galactose to
galactose-1-phosphate by galactokinase.
39
• Galactose-1-phosphate reacts with uridine diphosphate glucose
(UDP-glucose) to form UDP-galactose and glucose-1-phosphate.
• The reaction is catalyzed by galactose-1-phosphate uridyl
transferase.
40
• The galactose moiety of UDP-galactose is then epimerized
at carbon 4 to glucose by UDP-galactose-4-epimerase.
• Glucose-1-phosphate formed is then isomerized to
glucose-6-phosphate by phosphoglucomutase.
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Formation and Fate of Pyruvic acid (Pyruvate)
• Pyruvate is formed principally from the oxidation of glucose, via
the glycolysis pathway.
43
Formation of Pyruvic acid(Pyruvate) cont’d
• However, it can be formed from other sources which include;
• Conversion of lactic acid to pyruvic acid, catalyzed by lactate
dehydrogenase.
44
• Pyruvic acid can also be formed from oxaloacetate, either by
spontaneous decarboxylation or in a reaction catalyzed by
oxaloacetate decarboxylase.
45
• Pyruvic acid can be formed in the body from malic acid by malic
enzyme.
46
• Pyruvic acid is also formed from deamination of the amino acid
alanine.
• Certain glucogenic amino acids also produce pyruvic acid

during their catabolism e.g. glycine, serine, cysteine and
threonine.
47
Fate of Pyruvic acid (Formation of Acetyl CoA)
• The fate of pyruvic acid depends on the presence or
absence of oxygen.
• In the presence of oxygen, pyruvic acid undergoes
oxidative decarboxylation to form acetyl CoA.
• Pyruvate undergoes oxidative decarboxylation under
aerobic condition to form a 2-carbon compound ‘acetyl-
CoA’.
• The reaction involves both oxidation and
decarboxylation, and it is catalyzed by an multi-enzyme
complex known as pyruvate dehydrogenase complex,
which can exist both in the active and inactive forms.
48
Fate of Pyruvic acid (Formation of lactic acid)
• The Cori cycle
• Pyruvic acid is converted to lactic acid in the absence of oxygen via
the Cori or Lactic acid cycle.
• This is an important reaction and occurs in skeletal muscles, often
during periods of strenuous exercise.
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Fate of Pyruvic acid cont’d
• Pyruvic acid can also be converted to any of the following:
• Can be converted to alanine by amination
• Can form glucose via gluconeogenesis (this will be discussed
later)
• Forms malic acid which is in turn converted to OAA (oxaloacetic
acid)
• Can be converted to oxaloacetic acid (OAA) by CO2-fixation
reaction.
50
Gluconeogenesis
• Gluconeogenesis is the process by which glucose is
synthesized from non-carbohydrate sources.
• These non-carbohydrate sources include lactate,
glucogenic amino acids, glycerol and propionyl CoA
derived from odd chain fatty acids.
• Gluconeogenesis occurs in the liver.
• However, it also occur in the cells of the renal cortex to
a lesser extent.
• Some reactions of the gluconeogenetic pathway occur
in the cytosol while others occur in the mitochondria.
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Gluconeogenesis cont’d
• Most of the steps in the gluconeogenesis are a similar and
complete reverse of glycolysis with the same intermediates and
enzymes.
• However, the irreversible steps of glycolysis are circumvented by
four enzymes which are designated as the key enzymes of
gluconeogenesis.
• These key enzymes are:
• Pyruvate carboxylase
• Phosphoenol pyruvate carboxykinase
• Fructose-1-6-bisphosphatase
• Glucose-6-phosphatase
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• Pyruvate Carboxylase Reaction (Mitochondrial)
• Pyruvate formed in the cytoplasm enters the mitochondria with
the help of transfer proteins.
• Then, carboxylation of pyruvate to oxaloacetate occurs, being
catalyzed by a mitochondrial enzyme, pyruvate carboxylase that
requires the co-enzymes biotin and ATP.
54
• Phosphoenol Pyruvate Carboxy Kinase (Cytosolic)
• In the cytoplasm, PEPCK enzyme then converts oxaloacetate to
phosphoenol pyruvate, removing a molecule of CO2.
• GTP or ITP donates the phosphate.
• The net effect of these two reactions is the conversion of
pyruvate to phosphoenol pyruvate.
• This circumvents the irreversible step in glycolysis catalyzed by
pyruvate kinase (step 9 of glycolysis).
• The phosphoenol pyruvate soformed, undergoes further
reactions catalyzed by the glycolytic enzymes to form fructose-
1,6-bisphosphate (glycolysis steps 8,7,6,5 and 4). All these
reactions are freely reversible.
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• Fructose-1,6-bisphosphatase
• Fructose 1,6-bis-phosphate is then acted upon by
fructose 1,6-bisphosphatase to form fructose-6-
phosphate. This will bypass the step of PFK reaction (see
step 3 of glycolysis).
Fructose-1,6-bisphosphatase
Fructose-1,6-bisphosphate –––––––––→ Fructose-6-phosphate + Pi
• Then fructose-6-phosphate is isomerized to glucose-6-

phosphate by the freely reversible reaction catalyzed by
hexose phosphate isomerase (second step in glycolysis).
57
• Glucose-6-phosphatase Reaction
• The glucose 6-phosphate is hydrolyzed to free glucose by
glucose-6-phosphatase.
Glucose-6-phosphate + H2O -----→ Glucose + Pi
• Glucose-6-phosphatase is active in liver. It is present in kidney
and intestinal mucosa to a lesser extent, but is absent in
muscle.
58
• From Lactate
• Lactate is another substrate of gluconeogenesis. It is
often formed in muscles following anaerobic glycolysis.
• The lactate formed in the muscle is transported to the
liver.
• In the liver cell, lactate dehydrogenase converts it to
pyruvate.
• The pyruvate then enters the gluconeogenic pathway,
described above to form glucose.
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• From Glucogenic amino acids
• During periods of fasting, starvation or in diabetes
mellitus, when glucose is not readily available, the
glucogenic amino acids are transaminated to
corresponding carbon skeletons.
• These then enter the TCA cycle and form oxaloacetate or
pyruvate, which is then converted to glucose.
• Alanine released from the muscle is the major substrate
for gluconeogenesis.
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• From Glycerol
• The glycerol part of fat is phosphorylated in the liver
cytosol by ATP to glycerol-3-phosphate.
• The reaction is catalyzed by glycerol kinase.
• It is then oxidized to dihydroxyacetone phosphate
(DHAP) by an NAD+ dependent glycerol phosphate
dehydrogenase.
• DHAP then enters the gluconeogenic pathway to form
glucose.
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• Propionyl CoA
• The oxidation of odd chain fatty acids as well as the breakdown
of some amino acids (methionine, isoleucine) yields a three
carbon propionyl CoA.
• Propionyl CoA carboxylase acts on propionyl CoA in the
presence of ATP and biotin and converts it to methyl malonyl
CoA.
• Malonyl CoA is then converted to succinyl CoA in the presence
of Vit B12 coenzyme.
• Succinyl CoA formed from propionyl CoA enters
gluconeogenesis via the TCA cycle.
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Assignment
• Significance of gluconeogenesis
• Regulation of gluconeogenesis
67
Tricarboxylic acid (krebs) cycle
• The tricarboxylic acid (TCA) cycle is also known as the
citric acid or Krebs cycle.
• It was first called krebs cycle, being named after Sir Hans
Krebs who first proposed the complete cycle in 1937.
• He also named it TCA cycle.
• However, before him, some other scientists had
described some of the reactions in the cycle.
68
TCA cycle cont'd
• The TCA cycle is a sequence of reactions that occur in the
mitochondria of cells and involves the generation of energy from
the oxidation of acetyl CoA, which is derived from various
metabolic pathways of carbohydrates, fats and proteins.
• Acetyl CoA enters the TCA cycle and in the cycle becomes
completely oxidized, giving rise to carbon dioxide and energy.
• Pyruvate formed from glycolysis, undergoes oxidative
decarboxylation by pyruvate dehydrogenase and is converted to
acetyl CoA, which then enters the cycle.
• In the same vein, acetyl CoA is formed from other pathways,
thereby serving as a link between the TCA cycle and other
metabolic pathways.
• The cycle consists of eight(8) different reactions.
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Step 1: Formation of citrate
• In step 1, oxaloacetate is condensed with acetyl CoA in
an irreversible reaction, catalyzed by citrate synthase,
to form citrate (a tricarboxylic acid)
71
Step 2: Formation of Isocitrate
• Step 2 involves the isomerization of citrate to isocitrate
by the enzyme aconitase in a reversible reaction.
• The reaction occurs in two stages, with both catalyzed by
the same enzyme.
• The first stage involves the conversion of citrate to cis-
aconitase by the removal of one molecule of water.
• Cis-aconitase has short half life.
• Hence, it is immediately converted to isocitrate by
addition of one molecule of water.
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Step 3: Formation of alpha ketoglutarate
• In step 3, isocitrate undergoes oxidative decarboxylation
to alpha ketoglutarate with the liberation of one
molecule of carbon dioxide.
• The reaction is a two-stage reaction with both catalyzed
by isocitrate dehydrogenase.
• The first stage involves the dehydrogenation of Isocitrate
to oxalosuccinate.
• Oxalosuccinate is unstable and undergoes
decarboxylation to form alpha ketoglutarate.
• NADH is generated in this step, which enters the electron
transport chain (ETC) for the generation of ATPs.
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Step 4: Formation of succinyl CoA
• In step 4, there is oxidative decarboxylation of alpha
ketoglutarate to succinyl CoA by alpha ketoglutarate
dehydrogenase complex, which is a multi-enzyme
complex.
• The reaction is irreversible.
• NADH is also formed in this step, which enters the ETC
for the generation of ATPs.
• In this step too, another molecule of carbon dioxide is
liberated.
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Step 5: Formation of succinate
• Step 5 is a substrate level phosphorylation reaction in
which a molecule of GDP is phosphorylated to GTP and
succinate is formed.
• In this step, a high energy phosphate is generated from
the energy trapped in the thio-ester bond of succinyl
CoA.
• The GTP can be converted to ATP by reacting with an
ADP molecule:
• GTP + ADP → GDP + ATP
• Reaction is catalyzed by succinate thiokinase.
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Step 6: Formation of Fumarate
• Succinate is dehydrogenated to fumarate, an
unsaturated dicarboxylic acid, by succinate
dehydrogenase.
• The hydrogen atoms are received by FAD to form
FADH2, which then enters into ETC to generate ATPs.
• The enzyme, succinate dehydrogenase is
competitively inhibited by malonate.
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Step 7: Formation of Malate
• The formation of malate from fumarate is catalyzed by
fumarase.
• The reaction involves the addition of a water molecule.
• Only L-malate is formed.
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Step 8: Regeneration of Oxaloacetate
• In step 8, malate is oxidized to oxaloacetate by malate
dehydrogenase.
• NAD+ serves as coenzyme in this reaction.
• NADH is generated in this step, and it enters the
electron transport chain, to produce ATPs.
• The oxaloacetate can further condense with another
acetyl CoA molecule and the cycle continues.
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Regulation of TCA cycle
The TCA cycle is regulated by the following :
• Citrate synthase: This enzyme catalyses the formation of
citrate from oxaloacetate and acetyl CoA in step 1, and is
an important control.
• ATP is an allosteric inhibitor of this enzyme.
• Isocitrate dehydrogenase: This enzyme catalyses step 3
of the cycle.
• ADP is a positive modifier of this enzyme and enhances
the binding of substrate. NADH is an inhibitor.
87
Regulation of TCA cycle cont'd
• Alpha ketoglutarate dehydrogenase: Acts in step 4.
• It is inhibited by succinyl CoA and NADH.
• Availability and cellular need of ATP: When the energy charge of
the cell is low, the cycle operates at a faster rate.
• The cycle is tightly coupled to the respiratory chain providing
ATP.
• The Krebs cycle is the largest generator of ATP among metabolic
pathways.
• In anaerobic conditions, the cycle is inhibited.
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Assignment
• Highlight ten significance of the TCA cycle.
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Glycogenesis
• Glycogenesis is the process by which glycogen is formed
from glucose.
• Glycogen is a polysaccharide (homopolymer) deposited
in the tissues and stored as a carbohydrate.
• The primary purpose of glycogenesis is to make sure the
body does not run out of glucose.
• The pathway of glycogenesis includes a series of steps
that result in complex glycogen formation in the
cytoplasm of the liver and cells of the muscles.
90
Glycogenesis cont’d
• The steps of glycogenesis are as follows:
• Glucose phosphorylation – In the initial phase, glucose is
phosphorylated into glucose-6-phosphate, a usual reaction in
glycolysis.
• This reaction is catalyzed by glucokinase (liver) and hexokinase
(muscle).
• Formation of Glucose-1-phosphate – An enzyme
Phosphoglucomutase catalyzes the conversion of Glucose-6-
phosphate to Glucose-1-phosphate.
• Formation of UDP Glucose – The third step focuses on the
reaction of glucose-1-phosphate with UTP (uridine triphosphate)
to form the active nucleotide UDP-Glucose (Uridine diphosphate
glucose).
• The reaction is catalyzed by UDP-Glucose Pyrophosphorylase.
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• UDP-Glucose attaches to glycogen primer – A small fragment of
already existing glycogen serves as a primer, to stimulate the
synthesis of glycogen.
• The glucose from UDP-Glucose is accepted by glycogenin.
Glycogenin is an enzyme involved in converting glucose to
glycogen.
• It acts as a primer, by polymerizing the first few glucose
molecules, after which other enzymes take over.
• The initial glucose unit is attached to the hydroxyl group of
tyrosine of glycogenin, thereby acting as the primer.
• Other glucose molecules attach themselves to it.
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• Glycogen synthase synthesizes glycogen –Glycogen synthase
transfers glucose from UDP-Glucose to glycogen (non-reducing
end) forming alpha 1,4-linkages.
• The same enzyme catalyzes the synthesis of the unbranched
molecule with alpha-1,4-glycosidic linkages.
• Glycogen synthase synthesizes glycogen –Glycogen synthase
transfers glucose from UDP-Glucose to the non-reducing end of
glycogen forming alpha 1,4-linkages.
• The same enzyme catalyzes the synthesis of the unbranched
molecule with alpha-1,4-glycosidic linkages.
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• The formation of glycogen braches – The final step is the
formation of glycogen branches caused by the effect of branching
enzyme, which transfers a small fragment of about five to eight
residues of glucose from the non-reducing end of the glycogen
chain to another glucose residue linked by alpha-1,6 bond.
• This action causes the formation of a new non-reducing end.
• The final result is the elongation and branching out of the
glycogen chain.
• The process of glycogenesis utilizes two molecules of ATP.
• One molecule is needed for glucose phosphorylation and another
molecule is needed to convert UDP to UTP.
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Assignment
• Discuss the regulation of glycogenesis
• Discuss the significance of glycogenesis.
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Glycogenolysis
• Glycogenolysis refers to the breakdown of glycogen.
• Glycogenolysis is initiated by the action of a specific
enzyme phosphorylase, which brings about
Phosphorolytic cleavage of α 1→ 4 linkage to yield
glucose-1-phosphate.
• The liver and muscle contain the enzyme
phosphorylase.
• In both, the enzyme can be in ‘active’ and “inactive”
forms and both are interconvertible.
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Glycogenolysis cont’d
• Steps of Glycogenolysis
• Step 1: Action of Glycogen Phosphorylase:
• The alpha-1,4-glycosidic bonds (from non-reducing ends) are
cleaved sequentially by the enzyme Glycogen Phosphorylase to
yield Glucose-1-Phosphate.
• This process is called Phosphorolysis and it continues until four
glucose residues remain on either side of branching point (alpha-
1,6-glycosidic bond).
• The glycogen so formed is known as limit dextrin which can not
be further degraded by phosphorylase.
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• Step 2: Action of Debranching Enzyme:
• The branches of Glycogen are cleaved by two enzyme activities
present on a single polypeptide called debranching
enzyme, hence it is a bifunctional enzyme.
• Glycosyl 4:4 transferase (oligo alpha 1,4—>1,4 glucan
transferase) activity removes a fragment of three or four Glucose
residues attached at a branch and transfers them to another
chain.
• Here, one alpha-1,4-bond is cleaved and the same alpha-1,4
bond is made, but the places are different.
• Amylo alpha-1,6-glucosidase breaks the alpha-1,6-bond at the
branch with a single glucose residue.
101
• Step 3: Formation of Glucose-6-Phosphate and Glucose:
• Through the combined action of glycogen phosphorylase and
debranching enzyme, glucose-1-phosphate and free glucose in a
ration of 8:1 are produced.
• Glucose-1-phosphate is converted to glucose-6-phosphate by
the enzyme phosphoglucomutase.
• The fate of glucose-6-phosphate is as follows:
• In the liver, kidney and intestine (which have glucose-6-
phosphatase), glucose-6-phosphate is cleaved into glucose.
• The enzyme is absent in muscle and brain
• In peripheral tissues, glucose-6-phosphate will be used for
glycolysis.
102
Pentose phosphate pathway (ppp)
• The Pentose phosphate pathway is an alternative pathway for the
metabolism of glucose.
• It is also known as;
• Hexose monophosphate pathway or shunt
• Pentose cycle
• Phosphogluconate pathway
• Warburg-Dickens-Lipman Pathway.
• The pentose phosphate pathway is more complex than glycolysis.
• Like glycolysis, the reactions of the ppp occur in the cytosol.
103
Pentose Phosphate Pathway (PPP) cont’d
• Reactions of this pathway are considered in 2 stages.
• Stage I: Oxidative Phase
• The oxidative phase involves the oxidation of Glucose
and the formation of Pentose Phosphates.
• Stage II: Non-oxidation Phase
• The non-oxidative phase include:
• Pentose phosphate reactions (Interconversion of
pentoses) and
• Conversion of pentose phosphates to hexose
phosphates.
104
• Oxidative phase
• The oxidative phase involve the following reactions:
• Step 1: Dehydrogenation of glucose-6-phosphate to form 6-
phosphogluconolactone.
• The reaction is catalyzed by glucose 6-phosphate dehydrogenase.
• One molecule of NADPH is formed in the reaction.
• This is the rate limiting step of the reaction.
• Step 2: There is hydrolysis of 6-phosphogluconolactone to form 6-
phosphogluconate, catalyzed by gluconolactonase.
• Step 3: In this step, there is dehydrogenation of the 6-
phosphogluconate, followed by spontaneous decarboxylation to form
ribulose-5-phosphate, catalyzed by 6-phosphogluconate
dehydrogenase
• In this step, another molecule of NADPH is generated.
105
106
• Non-oxidative phase
• Step 4: Ribulose-5-phosphate is isomerized to the
corresponding ribose-5-phosphate by ribose-5-
phosphate isomerase or epimerized to xylulose-5-
phosphate by ribulose-5-phosphate 3-epimerase.
107
108
• Step 5: Xylulose-5-phosphate and ribose-5-phosphate reacts to
form glyceraldehyde-3-phosphate and a 7-carbon sugar,
sedoheptulose-7-phosphate by the enzyme transketolase.
• Transketolase is a TPP dependent enzyme.
109
• Step 6: There is a transfer of 3 carbon units from sedoheptulose-
7-phosphate to glyceraldehyde-3-phosphate to give fructose-6-
phosphate and erythrose-4-phosphate.
• The reaction is catalyzed by transaldolase
110
• Step 7: There is a transfer of 2 carbon units from xylulose-5-
phosphate to erythrose-4-phosphate to form fructose-6-
phosphate and glyceraldehyde-3-phosphate.
• The reaction is catalyzed by another transketolase which uses
Mg2+and thiamine diphosphate (vitamin B1) as coenzyme.
111
• Step 8: Two molecules of glyceraldehyde-3-phosphate
are condensed to form fructose-6-phosphate (reversal
of step 4 of glycolysis).
• Fructose-6 –phosphate is isomerized to glucose-6-
phosphate by enzyme phosphohexose isomerase
(reversal of step 2 of glycolysis).
112
113
Assignment
• Metabolic Role of NADPH formed by HMP Shunt
Pathway.
• Significance of the pentose phosphate pathway
• Regulation of the hexose monophosphate pathway
114
Electron transport chain (etc)
• The electron transport chain is the final common pathway
in aerobic cells by which electrons derived from various
substrates are transferred to oxygen.
• The energy rich carbohydrates (Glucose), Fatty acids and
amino acids undergo a series of metabolic reactions and
finally get oxidized to CO2 and H20, via the ETC.
• The reducing equivalents from various metabolic
intermediates are transferred to NAD+ and FAD to produce
NADH and FADH2.
• The reduced NADH and FADH2 pass through the ETC or
respiratory chain and finally reduce O2 to H20.
115
116
Electron transport chain cont’d
• The passage of electrons through the ETC is associated with the
loss of free energy, a part of which is utilized to generate ATP from
ADP and Pi.
• ETC is the 4th and final stage of aerobic respiration.
• Through ETC, the energy needed for cellular activities is released in
the form of ATP.
• ETC is an O2 dependent process which occurs in the inner
mitochondrial membrane.
• ETC is localized in Mitochondria.
• The mitochondria is the centre for metabolic oxidative reactions to
generate reduced coenzymes (NADH and FADH2) which in turn,
are utilized in ETC to liberate energy in the form of ATP.
• Hence, mitochondria is regarded as Power House of the Cell.
117
Electron transport chain cont’d
• The Mitochondria
• The mitochondrion consists of five distinct parts; the outer membrane, the
inner membrane, the intermembrane space, the cristae and the matrix.
• Mitochondrial matrix: The mitochondrial matrix is the site of the citric acid
cycle, beta-oxidation of fatty acids and oxidation of amino acids, as it is rich
in the enzymes responsible for the processes.
118
The Mitochondria cont’d
• Inner mitochondrial membrane : The inner
mitochondrial membrane is a specialized structure,
rich in proteins.
• It is impermeable to ions (H+, K+, Na+) and small
molecules (ADP, ATP).
• Cristae: The inner mitochondrial membrane is highly
folded to form cristae, which greatly increases its
surface area.
• Phosphorylating subunits: These are specialized
particles located in the inner surface of the inner
mitochondrial membrane and they are the centres for
ATP production.
119
The Mitochondria cont’d
• The inner mitochondrial membrane can be divided into five
distinct respiratory or enzyme complexes, denoted as complex I,
II, III, IV and V.
• The complexes l-lV are carriers of electrons while complex V is
responsible for ATP synthesis.
• Besides these enzyme complexes, there are certain mobile
electron carriers in the respiratory chain.
• These include NADH, coenzyme Q, cytochrome C and oxygen.
• The enzyme complexes (l-lV) and the mobile carriers are
collectively involved in the transport of electrons which,
ultimately, combine with oxygen to produce water.
120
Components of the ETC
• There are five distinct carriers that participate in the electron
transport chain (ETC).
• These carriers are responsible for the transfer of electrons from a
given substrate to ultimately combine with proton and oxygen to
form water.
• The carriers are:
• NAD & Flavoprotein: H+-carriers in cellular respiration
• Non heme metalloprotein (Fe-S- Protein): Iron cycles between 3+
and 2+ states.
• Ubiquinone or CoQ: region serves as an anchor to inner
mitochondrial membrane.
• Cytochromes : Electron-transfer proteins that contain a heme
prosthetic group
121
Composition of the Electron transport chain
• Four large protein complexes;
• Complex I - NADH-Coenzyme Q reductase
• Complex II - Succinate-Coenzyme Q reductase
• Complex III - Cytochrome c reductase
• Complex IV - Cytochrome c oxidase
• Many of the components are proteins with prosthetic groups
to move electrons.
122
123
Complex I
• It is also called NADH-CoQ reductase or NADH dehydrogenase
complex.
• NADH is the donor of electrons, which is accepted by FMN to
become reduced to FMNH2.
NADH + H+ + FMN → FMNH2 + NAD+
• The electrons from FMNH2 are then transferred to Fe-S and
then to coenzyme Q (ubiquinone) (CoQ).
• The overall function of this complex is to collect a pair of
electrons from NADH and pass them to CoQ.
• This creates a large negative free energy change, which is used
to drive 4 protons out of the mitochondria.
NADH → FMN → Fe-S → CoQ →
124
125
Complex II
• This is also known as Succinate-Q-Reductase complex.
• The electrons from FADH2 enter the ETC at the level of
coenzyme Q.
• Three major enzyme systems transfer their electrons directly to
ubiquinone from the FAD prosthetic group and they are:
• Succinate dehydrogenase
• Fatty acyl CoA dehydrogenase
• Mitochondrial glycerol phosphate dehydrogenase
• The ubiquinone (Q) accepts a pair of electrons from NADH or
FADH2 through complex-I or complex-II respectively, to become
reduced successively to semiquinone (QH) and finally to quinol
(QH2).
126
Complex III
• Also called cytochrome reductase complex.
• It is a cluster of iron-sulphur proteins, cytochrome b and
cytochrome c1, both contain heme prosthetic group.
• In the electron transfer process, the iron in heme group
shuttles between Fe3+ and Fe2+ forms, and 4 protons are
pumped out.
• Cytochrome c is a peripheral membrane protein
containing one heme prosthetic group.
• Cytochrome c collects electrons from Complex III and
delivers them to Complex IV.
127
128
Complex IV
• Also called cytochrome oxidase complex.
• It contains different proteins, including cytochrome a and cytochrome
a3.
• Four electrons are accepted from cytochrome c, and passed on to
molecular oxygen.
4H+ + O2 + 4Cyt.C-Fe++ → 2H2O + 4Cyt.C+++
• In this complex, 2 protons are pumped out to the intermembrane
space.
• Cytochrome oxidase has 4 redox centers, namely, a, a3, CuA and CuB,
and electron transfer is in the following order:
Cyto c →CuA→ Cyto a→ Cyto a3 →CuB
• Cytochrome oxidase contains 2 heme groups and 2 copper ions.
• The two heme groups are denoted as cytochrome-a and cytochrome
a-3.
• The functional unit of the enzyme is a single protein, and is referred to
as cytochrome a--a3
129
130
Complex V
• This is also known as ATP synthase complex.
• This is a protein assembly in the inner mitochondrial
membrane.
• Proton pumping ATP synthase (otherwise called F1-Fo
ATPase) is a multi subunit transmembrane protein.
• It has two functional units, named as F1 and Fo.
• Translocation of protons carried out by the Fo catalyzes
the formation of phospho-anhydride bond of ATP by F1.
• Coupling of the dissipation of proton gradient with ATP
synthesis (oxidative phosphorylation) is through the
interaction of F1 and Fo.
131
132
Oxidative Phosphorylation
• Oxidative phosphorylation is the process of
synthesizing ATP from ADP and Pi coupled with the
electron transport chain.
• The transport of electrons through the ETC is
linked with the release of free energy.
• The complex V of the inner mitochondrial
membrane is the site of oxidative phosphorylation.
133
P:O Ratio
• The P:O ratio refers to the number of molecules of ATP synthesized
per pair of electrons carried through the ETC.
• According to the estimated free energy of synthesis, it was
presumed that around 3 protons are required per ATP synthesized.
• When one NADH transfers its electrons to oxygen, 10 protons are
pumped out, while for FADH2, 6 protons are pumped out.
• This accounted for 3 ATP from NADH and 2 from FADH2.
• However, recent studies have shown that the actual energy
production is less, because there is always leakage of protons.
• Hence, this results in the production of only 2.5 ATP from NADH
and 1.5 ATP from FADH2.
• Therefore, when a pair of electrons from NADH reduces an atom of
oxygen (½ O2), 2.5 mol of ATP are formed per 0.5 mol of O2
consumed, while 1.5 mol for FADH2.
134
Convergence of metabolic pathways
• Metabolism is a continuous process, with thousands of
reactions, simultaneously occurring in the living cell.
• However, they are presented in form of pathways or
cycles for better understanding.
• The various pathways are interrelated as depicted in the
schematic diagram below.
• Glycolysis: This is responsible for the degradation of
glucose to pyruvate (lactate under anaerobic condition)
generating 8 ATP.
• Pyruvate is converted to acetyl CoA.
135
136
Convergence of metabolic pathways cont’d
• Citric acid cycle: Citric acid cycle is the final common metabolic
pathway for the oxidation of all foodstuffs.
• Most of the energy is trapped in the form of NADH and FADH2.
• Acetyl CoA, which is a common metabolite from different fuel
sources, is oxidized to CO2.
• Oxidative phosphorylation: The NADH and FADH2, produced in
different metabolic pathways, are finally oxidized in the electron
transport chain (ETC).
• The ETC is coupled with oxidative phosphorylation to generate
ATP.
• Gluconeogenesis: The synthesis of glucose from non-
carbohydrate sources constitutes gluconeogenesis.
• Several compounds (e.g. pyruvate, glycerol, amino acids) can
serve as precursors for gluconeogenesis
137
Convergence of metabolic pathways cont’d
• Pentose phosphate pathway: This pathway is primarily
concerned with the liberation of NADPH and ribose sugar.
• NADPH is utilized for the biosynthesis of several compounds,
including fatty acids.
• Ribose is an essential component of nucleotides and nucleic
acids.
• Glycogen metabolism: Glycogen is the storage form of glucose,
mostly found in liver and muscle.
• lt is degraded (glycogenolysis) and synthesized (glycogenesis) by
independent pathways.
• Glycogen effectively serves as a fuel reserve to meet body needs,
for a brief period (between meals)
138
Assignment
• Discuss the regulation of the electron transport
chain
• Discuss the significance of the electron transport
chain
139

Carbohydrate Metabolism

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Carbohydrate Metabolism

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Metabolism

• Certain glucogenic amino acids also produce pyruvic acid

Fructose-1,6-bisphosphate –––––––––→ Fructose-6-phosphate + Pi

• Then fructose-6-phosphate is isomerized to glucose-6-

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