1.carbohydrates and Lipid Metabolism-Converted - Watermark

Dr.
Thriveni V, Assistant Professor, AIT, Bnglr

METABOLISM OF CARBOHYDRATES AND LIPIDS
• Food: Complex mixture of carbohydrates, lipids, proteins
and vitamins.
• Food--→ carbon dioxide, water and energy.
Digestion
Glycolysis
Kreb’s cycle
• Digestion: complex polymeric macromolecules are broken-
down into small molecules. These molecules are transformed
to different cells
• Metabolism: The systematic and collective study of cell,
Enzymatic reactions (anabolic/catabolic) which takes place in
the cell. And the overall process is referred as metabolism.
• Two types of reactions
• Degradative reactions- Catabolism
• Synthetic reactions- Anabolism
Dr. Thriveni V, Assistant Professor, AIT, Bnglr
Stages in metabolism
1. Polymeric macromolecules degraded into small molecules
Carbohydrates→ polysaccharides→ Monosaccharides
Proteins→ polypeptides→ amino acids
Lipids→ Fats, oils→ Fatty acids
2. The product of digestion further degraded into still
simpler molecules.
Glycolysis
Urea cycle Acetyl
AcetylCo-A
Co-A
β-oxidation
3. Oxidation of the product of second stage into carbon
dioxide and water with the liberation of energy.
Occurs in mitochondria which is called as citric acid cycle
or Krebs cycle
Metabolism of carbohydrates
Starch
Starch
Salivary
amylase
Maltose
Pancreatic amylase
Maltose
Maltase
Glucose
Carbohydrate metabolism/ Glycolysis/EMP

cycle/EMP pathway
• Embeden-Meyerhof-Parnas (EMP) pathway
• Glycolysis is a set of reactions that takes place in the
cytoplasm of prokaryotes and eukaryotes.
• The roles of glycolysis is to produce energy (both directly
and by supplying substrate for the citric acid cycle
and oxidative phosphorylation) and to produce
intermediates for biosynthetic pathways.
• Glycolysis
• Aerobic glycolysis → Pyruvate.
• Anaerobic glycolysis→ pyruvate→ lactate.
• 10 steps in glycolysis.
• 2 phases
• Preparatory phase
• Pay-off phase
• Preparatory phase
• Includes 5 enzymatic reactions which converts 1
molecule of glucose into 2 molecules of
glyceraldehyde-3-phosphate
• 2 molecules of ATP are used.
• Pay-off phase
• 2 molecules of glyceraldehyde-3-phosphate gets
converted into 2 molecules of pyruvate
• 4 ATP molecules are produced.
• Net yield of two ATPs per molecule of glucose degraded.
1. Glucose is phosphorylated by ATP to form glucose 6-
phosphate and ADP. The reaction is catalyzed by the
enzyme glucokinase/hexokinase.
2. Glucose 6-phosphate is isomerised to fructose 6-phosphate by

glucose-6-phosphate isomerase/ phosphoglucoisomerase.
Isomerization involves the conversion of an aldose to a
ketose, a conversion that is better seen by viewing the
open chain representations of these molecules.
3. Fructose 6-phosphate is phosphorylated
Dr. Thriveni V,by ATPProfessor,
Assistant to form AIT, Bnglr
fructose 1,6-bisphosphate and ADP. The enzyme catalyzing

this step is phosphofructokinase (PFK).
4. Aldolase splits fructose 1,6-bisphosphate (a six-carbon

molecule) into two three-carbon molecules,
glyceraldehyde 3-phosphate and dihydroxyacetone
phosphate.
5. Glyceraldehyde 3-phosphate is the only molecule that can be

used for the rest of glycolysis. However, the
dihydroxyacetone phosphate formed in the previous step can
rapidly be converted to glyceraldehyde 3-phosphate by triose
phosphate isomerase.
This is an equilibrium reaction; as the glyceraldehyde 3-phosphate
is used by the rest of glycolysis, more dihydroxyacetone
phosphate is converted to glyceraldehyde 3-phosphate as
replacement. Thus effectively, for each molecule of fructose 1,6-
bisphosphate that is cleaved in step 4, two molecules of
glyceraldehyde 3- phosphate continue down the pathway.
6. Glyceraldehyde 3-phosphate is oxidised and phosphorylated to

1,3-bisphosphoglycerate. The reaction is catalyzed by
glyceraldehyde 3-phosphate dehydrogenase and uses
inorganic phosphate and NAD.
The other product is NADH. The energy for creating this new high-
energy phosphate bond comes from the oxidation of the
aldehyde group of the glyceraldehyde 3-phosphate.
7. The newly created high-energy phosphate bond of 1,3-
bisphosphoglycerate is now used to synthesize ATP.
Phosphoglycerate kinase catalyzes the transfer of the
phosphoryl group from the 1,3-bisphosphoglycerate to
ADP, generating ATP and 3-phosphoglycerate.
8. 3-Phosphoglycerate is isomerised to 2-phosphoglycerate by
phosphoglycerate mutase. Thus the reaction is a
movement of the phosphate group to a different carbon
atom within the same molecule.
9. Enolase catalyzes the dehydration of 2-phosphoglycerate to
form phosphoenolpyruvate (PEP). This reaction converts
the low-energy phosphate ester bond of 2-
phosphoglycerate into the high-energy phosphate bond of
PEP.
10. In the last reaction, pyruvate kinase catalyzes the
physiologically irreversible transfer of the phosphoryl group
from PEP to ADP to form ATP and pyruvate.
Aerobic glycolysis
Oxygen Presence
Oxidation of 32 mol of ATP per mol of glucose

glucose
Steps Occurs in 2 steps
1. Cytosol- 2 ATP + 2NADH
2. Mitochondrial oxidation: 30 ATP
Oxygen If it is in short
NADH is reoxidized
reduce pyruvate to lactate
This severely limits the amount of ATP formed per mole of
glucose oxidized
Imbalance of oxygen usage and oxygen delivery
Ex: Sepsis/heart failure
Anaerobic glycolysis occurs and results in lactate accumulation
and results in inefficient glucose usage and inadequate ATP
production.
Energy Yield in Aerobic Glycolysis
Step Enzyme Source No. of ATP
Formed/consu
med
1 Hexokinase – -1
3 Phosphofructokinase – -1
6 Glyceraldehyde-3- NADH (+3) x 2 = +6
phosphate
dehydrogenase
7 Phosphoglycerate kinase ATP (+1) x 2 = +2
10 Pyruvate kinase ATP (+1) x 2 = +2
Net Yield 8 ATPs
Other Monosaccharides Enter the Glycolytic Pathway at
Several Points
• In most organisms, hexoses other than glucose can undergo
glycolysis after conversion to a phosphorylated
derivatives.
• Fructose, present in free form in many fruits and formed
by hydrolysis of sucrose in the small intestine of
vertebrates, is phosphorylated by hexokinase:
• This is a major pathway of fructose entry into glycolysis in

the muscles and kidney. In the liver, however, fructose
enters by a different pathway. The liver enzyme
fructokinase catalyzes the phosphorylation of fructose
at C-1 rather than C-6:
• The fructose 1-phosphate is then cleaved to

glyceraldehyde and dihydroxyacetone phosphate by
fructose 1-phosphate aldolase:
• Dihydroxyacetone phosphate is converted to
glyceraldehyde 3-phosphate by the glycolytic enzyme
triose phosphate isomerase. Glyceraldehyde is
phosphorylated by ATP and triose kinase to
glyceraldehyde 3-phosphate
• D-Mannose, released in the digestion of various

polysaccharides and glycoproteins of foods, can be
phosphorylated at C-6 by hexokinase:
Mannose 6-phosphate is isomerized by phosphomannose

isomerase to yield fructose 6-phosphate, an
intermediate of glycolysis.
Fate of pyruvate
1. Conversion to lactate or anaerobic glycolysis
• When oxygen is not available NADH can not be oxidised
through ETC.
• In such cases Pyruvate is reduced to lactate by the
enzyme lactate dehydrogenase.
• Lactate can be reconverted into pyruvate when oxygen is
available.
• It occurs in the cytosol.
• Lactate is released by tissues (e.g., RBCs or exercising
muscle) and is used by the liver for gluconeogenesis
or by tissues such as the heart and kidney, where it is
converted to pyruvate and oxidized for energy.
• The LDH reaction is reversible.
• Different tissues have different mixtures of these

isozymes.
• Lactate dehydrogenase (LDH) consists of four subunits
that can be either of the muscle (M) or the heart (H)
type. Five isozymes occur (MMMM, MMMH, MMHH,
MHHH, and HHHH), which can be separated by
electrophoresis.
2. Conversion to ethanol or alcoholic fermentation
In some microorganisms like yeast when oxygen is not
available pyruvate is converted into ethanol and carbon
dioxide by the action of the enzymes pyruvate decarboxylase
and alcohol dehydrogenase. NADH is oxidized to NAD+
during this reaction
Pyruvate
Thiamine pyrophosphate
and Mg++. Pyruvate decarboxylase
Acetaldehyde + CO2
Alcohol dehydrogenase
Ethanol
3. Conversion to acetyl coenzyme A (CoA)
• In aerobic organisms pyruvate is oxidised to carbon
dioxide and water through citric acid cycle, ETC and
oxidative phosphorylation.
• Before the pyruvate enters into citric acid cycle it should
be converted into acetyl CoA
• Pyruvate can enter mitochondria, There it can be
converted by pyruvate dehydrogenase to acetyl CoA,
which can enter the TCA cycle.
• This reaction is catalysed by the enzyme pyruvate
dehydrogenase (PDH) complex.
Conversion of Pyruvate to acetyl coenzyme A (CoA)
∆ G= -33.4 kJ/mol
This step is the linking step between Glycolysis and TCA

cycle. This reaction leads to the formation of 6 ATP , 3
from each NADH molecule.
4. Conversion to oxaloacetate (OAA)
• Pyruvate can be converted to OAA by pyruvate
carboxylase.
• This reaction serves to replenish intermediates of
the TCA cycle as well as provide substrates for
gluconeogenesis.
• The enzyme is activated by acetyl CoA.
5. Conversion to alanine
• Pyruvate can be transaminated to form the amino
acid alanine.
• The enzyme involved is alanine aminotransferase,
which requires pyridoxal phosphate as a
cofactor.
Anaerobic glycolysis
• 10 seconds to 2 minutes
• The speed at which ATP is produced is about 100 times
that of oxidative phosphorylation.
• Lactate can be transformed by the liver back into glucose
through Cori cycle.
• Fates of pyruvate under anaerobic conditions:
Pyruvate is the terminal electron acceptor in lactic
acid fermentation.
• When sufficient oxygen is not present in the muscle cells
for further oxidation of pyruvate and NADH produced
in glycolysis, NAD+ is regenerated from NADH by
reduction of pyruvate to lactate.
• Pyruvate is converted to lactate by the enzyme lactate
dehydrogenase.
• The standard free energy change of the reaction is -25.1
kJ/mol.
Energy Yield in Anaerobic Glycolysis
Step Enzyme Source No. of ATP
Formed/con
sumed
1 Hexokinase – -1
3 Phosphofructokinase – -1
7 Phosphoglycerate ATP (+1) x 2 = +2
kinase
10 Pyruvate kinase ATP (+1) x 2 = +2
Net Yield 2 ATPs
Regulation ofDr.glycolysis
➢ Phosphofructokinase
The most important control step of glycolysis is the irreversible
reaction catalyzed by phosphofructokinase (PFK). The
enzyme is regulated in several ways:
● ATP/AMP. PFK is allosterically inhibited by ATP but this
inhibition is reversed by AMP. This allows glycolysis to
be responsive to the energy needs of the cell, speeding
up when ATP is in short supply (and AMP is plentiful) so
that more ATP can be made, and slowing down when
sufficient ATP is already available.
● Citrate. PFK is also inhibited by citrate, the first product of
the citric acid cycle. A high level of citrate signals that
there is a plentiful supply of citric acid cycle
intermediates already and hence no additional
breakdown of glucose via glycolysis is needed.
➢ Fructose 2,6-bisphosphate.
• Fructose 2,6-bisphosphate (F-2,6-BP) is synthesized from
fructose 6-phosphate by an enzyme called
phosphofructokinase.
• 2 (PFK2), a different enzyme from PFK. F-2,6-BP is
hydrolyzed back to fructose 6-phosphate by fructose
bisphosphatase 2 (FBPase2).
• Amazingly, both PFK2 and FBPase2 are activities catalyzed by
the same polypeptide; hence this is a bi-functional
enzyme.
• Fructose 6-phosphate stimulates the synthesis of F-2,6-BP
and inhibits its hydrolysis. F-2,6-BP in turn strongly
activates PFK and hence stimulates glycolysis.
• The overall effect is that when fructose 6-phosphate levels
are high, PFK (and hence glycolysis) is stimulated.
• When blood glucose levels fall, the hormone glucagon is

released into the bloodstream and triggers a cAMP
cascade that leads to phosphorylation of the
PFK2/FBPase2 polypeptide at a single serine residue.
• This activates FBPase2 and inhibits PFK2, lowering the level
of F-2,6-BP and hence decreasing the rate of glycolysis.
• The reverse is true as glucose levels rise; the phosphate
group is removed from the PFK2/FBPase2 polypeptide
by a phosphatase, thus inhibiting FBPase2 and
activating PFK2, raising the level of F-2,6-BP and hence
increasing the rate of glycolysis.
• Reciprocal regulation: F-2,6-BP is also important in
preventing glycolysis (glucose degradation) and
gluconeogenesis (glucose synthesis) operating
simultaneously.
● H ions. PFK is inhibited by H ions and hence the rate of
glycolysis decreases when the pH falls significantly.
This prevents the excessive formation of lactate (i.e.
lactic acid) under anaerobic conditions and hence
prevents the medical condition known as
acidosis (a deleterious drop in blood pH).
➢ Hexokinase
• Hexokinase, which catalyzes the first irreversible step
of glycolysis, is inhibited by glucose 6-phosphate.
• Thus when PFK is inhibited, fructose 6-phosphate builds up
and so does glucose 6-phosphate since these two
metabolites are in equilibrium via
phosphoglucoisomerase.
• The hexokinase inhibition then reinforces the inhibition
at the PFK step.
• At first sight this seems unusual since it is usually the first
irreversible step of a pathway (the committed step)
that is the main control step. On this basis, it may
appear that hexokinase should be the main control
enzyme, not PFK.
• However, glucose 6-phosphate, the product of the
hexokinase reaction, can also feed into glycogen
synthesis or the pentose phosphate pathway.
• Thus the first irreversible step that is unique to glycolysis is
that catalyzed by PFK and hence this is the main
control step.
➢ Pyruvate kinase
• Pyruvate kinase catalyzes the third irreversible step in
glycolysis. It is activated by fructose 1,6-
bisphosphate.
• ATP and the amino acid alanine allosterically inhibit the
enzyme so that glycolysis slows when supplies of
ATP and biosynthetic precursors (indicated by the
levels of Ala) are already sufficiently high.
• In addition, in a control similar to that for PFK, when the
blood glucose concentration is low, glucagon is
released and stimulates phosphorylation of the
enzyme via a cAMP cascade. This covalent
modification inhibits the enzyme so that glycolysis
slows down in times of low blood glucose levels.
Significance of the Glycolysis Pathway
• Glycolysis is the only pathway that is taking place in all the
cells of the body.
• It is the initial pathway by which glucose is used by the cell.
• Glycolysis is the only source of energy in erythrocytes.
• In strenuous exercise, when muscle tissue lacks enough
oxygen, anaerobic glycolysis forms the major source
of energy for muscles.
• The glycolytic pathway may be considered as the
preliminary step before complete oxidation.
• The glycolytic pathway provides carbon skeletons for
synthesis of non-essential amino acids as well as
glycerol part of fat.
• Most of the reactions of the glycolytic pathway are
reversible, which are also used for gluconeogenesis.
• It produces intermediates like acetyl CoA which is a

precursor for many biosynthetic pathways.
• Sugars other than glucose enter glycolytic pathway and
then converted into intermediates of glycolysis to enter
into glycolysis pathway.
• It leads the formation of ATP.
• The cycle forms the central part of a three-step process
Step 1 – Oxidation of fuel molecules to acetyl CoA
A major source of energy is glucose which is converted
by glycolysis into pyruvate. Pyruvate dehydrogenase (a
complex of three enzymes and five coenzymes) then oxidizes
the pyruvate (using NAD which is reduced to NADH) to form
acetyl CoA and CO2. Since the reaction involves both an
oxidation and a loss of CO2, the process is called oxidative
decarboxylation.
Step 2 – The citric acid cycle
The cycle carries out the oxidation of acetyl groups
from acetyl CoA to CO2 with the production of four pairs of
electrons, stored initially in the reduced electron carriers
NADH and FADH2
Step 3 – Oxidation of NADH and FADH2 produced by the

citric acid cycle
The NADH and FADH2 produced by the citric acid cycle are
re-oxidized and the energy released is used to synthesize ATP
by oxidative phosphorylation.
Tricarboxylic acid (TCA) cycle/ Krebs cycle/ Citric acid
cycle
Hans Kreb in the year 1937
Role
• The cycle oxidizes pyruvate to CO2 and H2O, with the
concomitant production of energy.
• Acetyl CoA from fatty acid breakdown and amino acid
degradation products are also oxidized.
• Cycle has a role in producing precursors for biosynthetic
pathways.
Location
• Mitochondria of eukaryotes.
Cytosol of prokaryotes.
Succinate dehydrogenase, the only membrane-bound enzyme in the
citric acid cycle, is embedded in the inner mitochondrial membrane
in eukaryotes and in the plasma membrane in prokaryotes.
All the enzymes of citric acid cycle are located inside the
mitochondria.
1. Citrate (6C) is formed from the irreversible condensation of
acetyl CoA (2C) and oxaloacetate (4C) – catalyzed by
citrate synthase.
2. Citrate is converted to isocitrate (6C) by an isomerization
reaction catalyzed by aconitase.
This is actually a two-step reaction during which cis-
aconitate is formed as an intermediate. It is the cis-
aconitate which gives the enzyme its name.
3. Isocitrate is oxidized to α-ketoglutarate (5C) and CO2 by
isocitrate dehydrogenase.
This mitochondrial enzyme requires NAD, which is reduced
to NADH.
4. α-Ketoglutarate is oxidized to succinyl CoA (4C) and CO2 by
the α-ketoglutarate dehydrogenase complex. Like
pyruvate dehydrogenase, this is a complex of three
enzymes and uses NAD as a cofactor.
5. Succinyl CoA is converted to succinate (4C) by succinyl CoA
synthetase. The reaction uses the energy released by
cleavage of the succinyl–CoA bond to synthesize either
GTP (mainly in animals) or ATP (exclusively in plants)
from Pi and, respectively, GDP or ADP.
6. Succinate is oxidized to fumarate (4C) by succinate
dehydrogenase. FAD is tightly bound to the enzyme
and is reduced to produce FADH2.
7. Fumarate is converted to malate (4C) by fumarase; this is a
hydration reaction requiring the addition of a water
molecule.
8. Malate is oxidized to oxaloacetate (4C) by malate
dehydrogenase. NAD is again required by the enzyme
as a co-factor to accept the free pair of electrons and
produce NADH.
Energy yield from TCA cycle

• Each of the three NADH molecules produced per turn of
the cycle yields 3 ATPs. (1NADH = 3 ATP)
• 3 NADH = 3 x 3 = 9 ATP molecules
• 1 GTP = 1 ATP conversion of succinyl CoA to succinate
• 1 FADH2 = 2 ATP (oxidative phosphorylation)
Total = 12 ATP molecules
(NADH and FADH2: although some measurements indicate

that the quantities are 2.5 and 1.5 respectively)
• Thus the oxidation of a single molecule of acetyl CoA via

the citric acid cycle produces 12 ATP molecules.
Regulation of the Citric Acid Cycle
• The overall rate of the citric acid cycle is controlled by the

rate of conversion of pyruvate to acetyl-CoA.
• BY the flux through citrate synthase, isocitrate
dehydrogenase, and α-ketoglutarate dehydrogenase.
These fluxes are largely determined by the
concentrations of substrates and products: the end
products ATP and NADH are inhibitory, and the
substrates NAD and ADP are stimulatory.
Functions of the Citric Acid Cycle
1. It is the final common oxidative pathway that oxidizes
acetyl CoA to CO2.
2. It is the source of reduced coenzymes that provide the
substrate for the respiratory chain.
3. Citric acid cycle is an amphibolic pathway, It acts as a link
between catabolic and anabolic pathways. Besides its
role in the oxidative catabolism of carbohydrates, fatty
acids, and amino acids, the cycle provides precursors
for many biosynthetic pathways.
4. It provides precursors for synthesis of amino acids and
nucleotides.
5. Components of the cycle have a direct or indirect
controlling effect on key enzymes of other pathways.
Summary of ATP yield from complete aerobic oxidation of
glucose
Glycolysis = 8 ATP
Pyruvate to acetyl CoA = 6 ATP (3 from each)
TCA cycle = 24 ATP (12 from each acetyl CoA)
Total = 38 ATP molecules.

Glyoxylate Cycle
• Kornberg and Kreb’s in the year 1957 framed a cycle called
as Glyoxylic acid cycle or Glyoxylate Cycle
• The pathway through which the fats could be converted into
sucrose (i.e., carbohydrate) during the germination of
fatty seeds in plants.
• The glyoxylate cycle is intimately associated with Kreb’s
Cycle.
• It is now known to occur in many other bacteria, yeasts,
molds, and higher plants and is completed in
glyoxysomes, mitochondria and cytosol.
Steps
• Acetyl-CoA condenses with oxaloacetate to form citrate.
• Citrate is converted to isocitrate, exactly as in the citric
acid cycle.
• The next step, however, is not the breakdown of isocitrate by
isocitrate dehydrogenase but the cleavage of isocitrate
by isocitrate lyase, forming succinate and glyoxylate.
• The glyoxylate then condenses with a second molecule
of acetyl-CoA to yield malate, in a reaction catalyzed by
malate synthase.
• The malate is subsequently oxidized to oxaloacetate, this
was catalysed by malate dehydrogenase.
• Which can condense with another molecule of acetyl-CoA to
start another turn of the cycle
• Each turn of the glyoxylate cycle consumes two molecules
of acetyl-CoA and produces one molecule of succinate,
which is then available for biosynthetic purposes.
• The succinate may be converted through fumarate and
malate into oxaloacetate, which can then be converted
to phosphoenolpyruvate by PEP carboxykinase, and
thus to glucose by gluconeogenesis.
• Vertebrates do not have the enzymes specific to the
glyoxylate cycle (isocitrate lyase and malate synthase)
and therefore cannot bring about the net synthesis of
glucose from lipids.
• In plants, the enzymes of the glyoxylate cycle are
sequestered in membrane-bounded organelles called
glyoxysomes, which are specialized peroxisomes.
• Glyoxysomes are not present in all plant tissues at all times.

They develop in lipid-rich seeds during germination,
before the developing plant acquires the ability to make
glucose by photosynthesis.
• In addition to glyoxylate cycle enzymes, glyoxysomes
contain all the enzymes needed for the degradation of
the fatty acids stored in seed oils.
• Acetyl-CoA formed from lipid breakdown is converted to
succinate via the glyoxylate cycle, and the succinate is
exported to mitochondria, where citric acid cycle
enzymes transform it to malate.
• Malate is oxidizes to oxaloacetate catalysed by a cytosolic
enzyme malate dehydrogenase.
• Oxaloacetate is a precursor for gluconeogenesis.
• Germinating seeds can therefore convert the carbon of
stored lipids into glucose.
Relationship between the

glyoxylate and citric acid cycles.
Coordinated regulation of glyoxylate
Dr. and
Thrivenicitric acid
V, Assistant cycles.
Professor, AIT, Bnglr
• Regulation of isocitrate
dehydrogenase activity
determines the partitioning
of isocitrate between the
glyoxylate and citric acid
cycles.
• When the enzyme is inactivated by
phosphorylation (by a
specific protein kinase),
isocitrate is directed into
biosynthetic reactions via the
glyoxylate cycle.
• When the enzyme is activated by
dephosphorylation
(by a specific phosphatase),
isocitrate enters the citric
acid cycle and ATP is
produced.
Pentose phosphateDr.pathway
• Reducing power is available in a cell both as NADH and

NADPH but these have quite distinct roles.
• NADH is oxidized by the respiratory chain to generate
ATP via oxidative phosphorylation.
• NADPH is used for biosynthetic reactions that require
reducing power.
Nicotinamide adenine
dinucleotide exists in
two forms.
• Oxidized form NAD+
• Reduced form NADH
Nicotinamide adenine
dinucleotide phosphate
exists in two forms.
• Oxidized form NADP+
• Reduced form NADPH.
• NADH and NADPH are not metabolically interchangeable
and so the cell must carry out a set of reactions that
specifically create NADPH.
• This set of reactions is the pentose phosphate pathway
• It takes place in the cytosol and is particularly important
in tissues such as adipose tissue, mammary gland and
the adrenal cortex that synthesizes fatty acids and
steroids from acetyl CoA.
• The activity of the pathway is very low in skeletal muscle,
(ex: which does not synthesize fatty acids or steroids).
Core reaction of the pathway
The pathway has three stages:

Stage 1 – Oxidative reactions that convert glucose 6-
phosphate into ribulose 5-phosphate, generating two
NADPH molecules
Stage 2 – Isomerization of ribulose 5-phosphate to ribose 5-
phosphate
Stage 3 – Linkage of the pentose phosphate pathway to
glycolysis via transketolase and transaldolase
Stage 1 – Oxidative reactions
• Glucose 6-phosphate is oxidized by Glucose 6-phosphate
dehydrogenase to 6-phosphoglucono-δ-lactone (producing
NADPH)
• 6-phosphoglucono-δ-lactone hydrolyzed by Lactonase to give 6-
phosphogluconate.
• The 6-phosphogluconate is subsequently converted by 6-
phosphogluconate dehydrogenase to ribulose 5-phosphate.
• This is an oxidative decarboxylation (i.e. the 6-phosphogluconate
is oxidized and a carbon is removed as CO2).
Stage 2 – Isomerization
• The ribulose 5-phosphate is converted to ribose 5-
phosphate by isomerization, a reaction catalyzed by
phosphopentose isomerase
Stage 3 – Linkage of the pentose phosphate pathway to
glycolysis
• If
at any time only a little ribose
5-phosphate is required
for nucleic acid
synthesis and other
synthetic reactions, it
will tend to accumulate
and is then converted to
fructose 6-phosphate
and glyceraldehyde 3-
phosphate by the
enzymes transketolase
and transaldolase.
• These two products are
intermediates of
glycolysis.
Regulation of Pentose phosphate pathway
• The transketolase and transaldolase reactions are reversible,
so the final products of the pentose phosphate pathway
can change depending on the metabolic needs of the
cell.
• When the cell needs NADPH but not ribose 5-phosphate,
the latter is converted to glycolytic intermediates and
enters glycolysis.
• At the other extreme, when the need for ribose 5-phosphate
exceeds that for NADPH, fructose 6-phosphate and
glyceraldehyde 3-phosphate can be taken from glycolysis
and converted into ribose 5-phosphate by reversal of the
transketolase and transaldolase reactions.
➢ Glucose 6-phosphate dehydrogenase

• The first reaction of the pathway, the oxidation of glucose
6-phosphate by glucose 6-phosphate dehydrogenase, is
rate limiting and irreversible.
• The enzyme is regulated by NADP. As the cell uses NADPH,
the concentration of NADP rises, stimulating glucose 6-
phosphate dehydrogenase and so increasing the rate of
the pathway and NADPH regeneration.
Electron transport chain (ETC) and oxidative

phosphorylation
• Oxidative phosphorylation is by far the major source of ATP
in the cell.
• These processes re-oxidize the NADH and FADH2 that arise
from
▪ The citric acid cycle (located in the mitochondrial
matrix)
▪ Glycolysis (located in the cytoplasm)
▪ Fatty acid oxidation (located in the mitochondrial
matrix)
• Trap the energy released as ATP.
• In eukaryotes, electron transport and oxidative

phosphorylation occur in the inner membrane of
mitochondria.
• In prokaryotes, the components of electron transport
and oxidative phosphorylation are located in the
plasma membrane.
Redox potential
• The oxidation of a molecule involves the loss of electrons.
• The reduction of a molecule involves the gain of electrons.
• Since electrons are not created or destroyed in a chemical
reaction, if one molecule is oxidized, another must be
reduced (i.e. it is an oxidation–reduction reaction).
• Oxidation–reduction reactions involve the transfer of
electrons. In the oxidation–reduction reaction:
• When the NADH is oxidized to NAD, it loses electrons. When

the molecular oxygen is reduced to water, it gains
electrons.
• The oxidation–reduction potential, E, (or redox potential)
is a measure of the affinity of a substance for electrons
and is measured relative to hydrogen.
• Positive redox potential: The substance has a higher affinity
for electrons than does hydrogen and so would accept
electrons from hydrogen.
• Negative redox potential: The substance with lower affinity
for electrons than does hydrogen and would donate
electrons to H, forming hydrogen.
• NADH is a strong reducing agent with a negative redox

potential and has a tendency to donate electrons.
• Oxygen is a strong oxidizing agent with a positive redox

potential and has a tendency to accept electrons.
Energetics
• For biological systems, the standard redox potential for a
substance (E0’) is measured under standard conditions,
at pH 7, and is expressed in volts.
• In an oxidation–reduction reaction, where electron transfer
is occurring, the total voltage change of the reaction
(change in electric potential, ΔE) is the sum of the
voltage changes of the individual oxidation–reduction
steps.
• The standard free energy change of a reaction at pH 7,
ΔG0′, can be readily calculated from the change in
redox potential ΔE0′ of the substrates and products:
Electron transport chain/ ETC/ Respiratory chain/
Oxidative phosphorylation
• Most of the protein components of the electron transport
chain exist as four large protein complexes embedded in
the inner mitochondrial membrane called:
• NADH-CoQ reductase (Complex I)
• Succinate-Q reductase (Complex II)
• Q-cytochrome c reductase (Complex III)
• Cytochrome c oxidase (Complex IV)
• Electron carriers are arranged such that the electron
accepted from NADH and FADH2 flows down a free
energy gradient to oxygen.
• Electron flow through ETC doesnot directly lead to ATP
synthesis. At certain points proton translocation
takesplace across the membrane creating the proton
gradient. This is used for the synthesis of ATP.
Complex 1: it contains FMN as cofactor. It accepts 2 electrons

from NADH in the form of hydride ion (H-). FMN is
reduced to FMNH2 then passes on the electron to
Coenzyme Q (CoQ) which is converted into CoQH2.
Complex 2: it contains FAD as prosthetic group. It accepts
electrons from the substrate and get converted into
FADH2. the electrons are then transferred to coenzyme
Q reducing it to CoQH2. Thus coenzyme Q accepts
electrons from both the complex 1 and 2. it is then
transferred to complex 3.
Complex 3: this complex contains cytochrome b and c1. it
transfers the electron from CoQH2 to cytochrome c.
Cytochrome c transfers electron from complex 3 to
complex 4.
Complex 5: This complex contains cytochrome a and
cytochrome a3. it catalyses complex reaction of transfer
of 4 electrons from cytochrome c along with 4 protons
from the surrounding medium to molecular oxygen.
Oxidative phosphorylation/ Synthesis of ATP
• It occurs when NADH and FADH2 are oxidized (hence
oxidative) by electron transport through the respiratory
chain.
• Unlike substrate level phosphorylation, it does not involve
phosphorylated chemical intermediates.
• Oxidative phosphorylation mechanism was proposed by
Peter Mitchell in 1961, the chemiosmotic hypothesis.
• This proposes that energy liberated by electron transport is
used to create a proton gradient across the
mitochondrial inner membrane and that is used to drive
ATP synthesis.
• Thus the proton gradient couples electron transport and ATP
synthesis, not a chemical intermediate.
• The evidence is overwhelming that this is indeed the
way that oxidative phosphorylation works. The actual
synthesis of ATP is carried out by an enzyme called ATP
synthase located in the inner mitochondrial
membrane.
Difference between Oxidative phosphorylation and
substrate level phosphorylation
Sl Oxidative Substrate level
No phosphorylation phosphorylation
1 Occurs in Inner mitochondrial Cytoplasm/ mitochondrial
membrane matrix
2 Enzyme ATP synthase Various kinase enzymes
system
3 Requirement Oxygen as final electron Does not require any
acceptor acceptor
4 Source of Proton gradient Energy released by the
energy generated during ETC is hydrolysis of high energy
conserved by the compounds is used for the
phosphorylation of ADP formation of ATP from ADP
to ATP and Pi
Coupling and respiratory control
• Electron transport is normally tightly coupled to ATP
synthesis (i.e. electrons do not flow through the
electron transport chain to oxygen unless ADP is
simultaneously phosphorylated to ATP).
• ATP is not synthesized unless electron transport is occurring
to provide the proton gradient.
• Thus oxidative phosphorylation needs NADH or FADH2,
oxygen, ADP and inorganic phosphate.
• The actual rate of oxidative phosphorylation is set by the
availability of ADP. If ADP is added to mitochondria, the
rate of oxygen consumption rises as electrons flow
down the chain and then the rate of oxygen utilization
falls when all the ADP has been phosphorylated to ATP;
a process called respiratory control.
• This mechanism ensures that electrons flow down the
chain only when ATP synthesis is needed. If the level
of ATP is high and the ADP level is low, no electron
transport occurs, NADH and FADH2 build up, as does
excess citrate, and the citric acid cycle and glycolysis
are inhibited.
Electron transport Inhibitors

• Several inhibitors of specific electron carriers are known
and were used in the original studies to determine the
order of the components in the respiratory chain.
• Rotenone and amytal inhibit electron transport at NADH-Q
reductase and so prevent NADH oxidation but the
oxidation of FADH2 can still occur since this feeds
electrons into the chain at CoQ.
• Antimycin A inhibits electron transport at the Q-
cytochrome c reductase complex.
• Cyanide (CN–), azide (N3–) and carbon monoxide (CO) all
inhibit cytochrome c oxidase.
Oxidative stress
• A free radical is a molecule or molecular fragment that
contains one or more unpaired electrons in its outer
orbital.
• Free radical is generally represented by a superscript dot,
(R• ).
• Oxidation reactions ensure that molecular oxygen is
completely reduced to water. The products of partial
reduction of oxygen are highly reactive and create havoc
in the living systems. Hence, they are also called
Reactive oxygen species or ROS.
Members of ROS:
• Superoxide anion radical (O2--• )
• Hydroperoxyl radical (HOO• )
• Hydrogen peroxide (H2O2)
• Hydroxyl radical (OH•)
• Lipid peroxide radical (ROO•)
• Singlet oxygen ( 1O2)
• Nitric oxide (NO•)
• Peroxy nitrite (ONOO--•).
Characteristics of ROS:
a. Extreme reactivity.
b. Short life span.
c. Generation of new ROS by chain reaction.
d. Damage to various tissues.
• Oxidative stress is a phenomenon caused by an imbalance
between production and accumulation of oxygen
reactive species (ROS) in cells and tissues and the ability
of a biological system to detoxify these reactive
products.
• ROS are generated in biological systems by aerobic
metabolism and also by exogenous sources such as
drugs, ultraviolet light, ionizing radiation and pollution
systems.
• Many endogenous and exogenous defense mechanisms are
available in living organisms to limit the levels of ROS
and the damage caused by these radicals.
• The defense mechanisms include antioxidant enzymes

such as superoxide dismutase, catalase, glutathione
peroxidase and many large non-enzymatic antioxidant
compounds such as, polyphenols, tocopherols, ascorbic
acid, uric acid, glutathione and other thiol protein
groups to protect the functional and structural integrity
of the biological molecules.
• Unbalanced ROS production and antioxidant cell defenses
have been associated in the physiological and
pathological conditions such as Cardiovascular Disease,
asthma and chronic obstructive pulmonary disease,
aging, cancer, rheumatoid arthritis, atherosceloresis
and neurodegenerative diseases including Alzheimer's
and Parkinson's etc.
Gluconeogenesis
• Gluconeogenesis synthesizes glucose from non-carbohydrate
precursors, including lactate and pyruvate, citric acid
cycle intermediates, the carbon skeletons of most amino
acids and glycerol.
• The main site of gluconeogenesis is the liver, although it also
occurs to a far lesser extent in the kidneys. Very little
gluconeogenesis occurs in brain or muscle.
• Within liver cells, the first enzyme of gluconeogenesis,
pyruvate carboxylase, is located in the mitochondrial
matrix. The last enzyme, glucose 6-phosphatase is
bound to the smooth endoplasmic reticulum. The other
enzymes of the pathway are located in the cytosol.
• Gluconeogenesis appears to be a reversal of glycolysis.
Comparison of
gluconeogenesis and glycolysis
Precursors for gluconeogenesis Dr. Thriveni V, Assistant Professor, AIT, Bnglr
• Glycerol can act as a substrate for glucose synthesis by

conversion to dihydroxyacetone phosphate, an
intermediate in gluconeogenesis.
• For lactate, pyruvate, citric acid cycle intermediates and the
carbon skeletons of most amino acids to act as
precursors for gluconeogenesis, these compounds
must first be converted to oxaloacetate.
• Some of the carbon skeletons of the amino acids directly
give rise to oxaloacetate. Others feed into the citric acid
cycle as intermediates and the cycle then converts these
molecules to oxaloacetate.
• Lactate is converted to pyruvate by the lactate dehydrogenase
reaction and some amino acids also give rise to pyruvate.
• Therefore, for these precursors, the first step in the
gluconeogenic pathway is the conversion of pyruvate to
oxaloacetate.
Steps in gluconeogenesis
1. Pyruvate is converted to oxaloacetate by carboxylation using
the enzyme pyruvate carboxylase that is located in the
mitochondrial matrix.
This enzyme uses biotin as an activated carrier of CO2, the

reaction occurring in two stages:
2. The oxaloacetate is now acted on by phosphoenolpyruvate
carboxykinase which simultaneously decarboxylates
and phosphorylates it to form phosphoenolpyruvate
(PEP), releasing CO2 and using GTP in the process
3. PEP is converted to fructose 1,6-bisphosphate in a series of
steps that are a direct reversal of those in glycolysis,
using the enzymes enolase, phosphoglycerate mutase,
phosphoglycerate kinase, glyceraldehyde 3-phosphate
dehydrogenase, triose phosphate isomerase and
aldolase. This sequence of reactions uses one ATP and
one NADH for each PEP molecule metabolized
4. Fructose 1,6-bisphosphate is dephosphorylated to form

fructose 6-phosphate by the enzyme fructose 1,6-
bisphosphatase.
5. Fructose 6-phosphate is converted to glucose 6-phosphate
by the glycolytic enzyme phosphoglucoisomerase.
6. Glucose 6-phosphate is converted to glucose by the enzyme

glucose 6-phosphatase. This enzyme is bound to the
smooth endoplasmic reticulum.
Energy used
• The synthesis of glucose by gluconeogenesis needs the
input of energy.
• Two pyruvate molecules are required to synthesize one
molecule of glucose.
• Energy is required at the following steps:
Pyruvate carboxylase 1 ATP (× 2) = 2 ATP
PEP carboxykinase 1 GTP (× 2) = 2 ATP
Phosphoglycerate kinase 1 ATP (× 2) = 2 ATP
Total = 6 ATP
• The net ATP yield from glycolysis is 2.
• Thus an extra 4 ATPs/ glucose are required to reverse
glycolysis.
• The glyceraldehyde 3-phosphate dehydrogenase reaction
also consumes NADH.
• 1NADH = 3 ATP molecules.
• Since each cytosolic NADH would normally be used to
generate approximately 3 ATP molecules via the
glycerol 3-phosphate shuttle and oxidative
phosphorylation, this is equivalent to the input of
another 6 ATPs per glucose synthesized.
Reciprocal regulation of glycolysis and Gluconeogenesis
• Glycolysis generates two ATPs net per glucose.
• Gluconeogenesis uses four ATPs and two GTPs per glucose.
• Operation of both glycolysis and gluconeogenesis, the net
result would be the utilization of two ATPs and two
GTPs, a so-called futile cycle.
• This is prevented by tight coordinate regulation of
glycolysis and gluconeogenesis. Since many of the steps
of the two pathways are common, the steps that are
distinct in each pathway are the sites of this regulation,
in particular the interconversions between fructose 6-
phosphate and fructose 1,6-bisphosphate and
between PEP and pyruvate
Regulation of pyruvate kinase, pyruvate carboxylase and
PEP carboxykinase
• In liver, pyruvate kinase is inhibited by high levels of ATP and
alanine so that glycolysis is inhibited when ATP and
biosynthetic intermediates are already plentiful.
• Acetyl CoA is also abundant under these conditions and
activates pyruvate carboxylase, favoring
gluconeogenesis.
• When the energy status of the cell is low, the ADP
concentration is high and this inhibits both pyruvate
carboxylase and PEP carboxykinase, switching off
gluconeogenesis. At this time, the ATP level will be low
so pyruvate kinase is not inhibited and glycolysis will
operate.
• Pyruvate kinase is also stimulated by fructose 1,6-

bisphosphate, so that its activity rises when needed, as
glycolysis speeds up.
• During starvation, the priority is to conserve blood glucose
for the brain and muscle. Under these conditions,
pyruvate kinase in the liver is switched off. This occurs
because the hormone glucagon is secreted into the
bloodstream and activates a cAMP cascade that leads
to the phosphorylation and inhibition of this enzyme.
Biosynthesis of Fatty acids

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Dr.

Thriveni V, Assistant Professor, AIT, Bnglr

Carbohydrate metabolism/ Glycolysis/EMP

2. Glucose 6-phosphate is isomerised to fructose 6-phosphate by

fructose 1,6-bisphosphate and ADP. The enzyme catalyzing

4. Aldolase splits fructose 1,6-bisphosphate (a six-carbon

5. Glyceraldehyde 3-phosphate is the only molecule that can be

6. Glyceraldehyde 3-phosphate is oxidised and phosphorylated to

Oxidation of 32 mol of ATP per mol of glucose

• This is a major pathway of fructose entry into glycolysis in

• The fructose 1-phosphate is then cleaved to

• D-Mannose, released in the digestion of various

Mannose 6-phosphate is isomerized by phosphomannose

• Different tissues have different mixtures of these

This step is the linking step between Glycolysis and TCA

• When blood glucose levels fall, the hormone glucagon is

• It produces intermediates like acetyl CoA which is a

Step 3 – Oxidation of NADH and FADH2 produced by the

Energy yield from TCA cycle

(NADH and FADH2: although some measurements indicate

• Thus the oxidation of a single molecule of acetyl CoA via

Regulation of the Citric Acid Cycle

• The overall rate of the citric acid cycle is controlled by the

Total = 38 ATP molecules.

• Glyoxysomes are not present in all plant tissues at all times.

Relationship between the

• Reducing power is available in a cell both as NADH and

The pathway has three stages:

➢ Glucose 6-phosphate dehydrogenase

Electron transport chain (ETC) and oxidative

• In eukaryotes, electron transport and oxidative

• When the NADH is oxidized to NAD, it loses electrons. When

• NADH is a strong reducing agent with a negative redox

• Oxygen is a strong oxidizing agent with a positive redox

Complex 1: it contains FMN as cofactor. It accepts 2 electrons

Electron transport Inhibitors

• The defense mechanisms include antioxidant enzymes

• Glycerol can act as a substrate for glucose synthesis by

This enzyme uses biotin as an activated carrier of CO2, the

4. Fructose 1,6-bisphosphate is dephosphorylated to form

6. Glucose 6-phosphate is converted to glucose by the enzyme

• Pyruvate kinase is also stimulated by fructose 1,6-

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