Metabolism

NAT SCI 9 BIOCHEMISTRY
Module 3
Metabolism
Module Overview:
Metabolism is the sum of all chemical reactions needed to sustain life. Living
organisms are unique in that they can extract energy from their environments
and use it to carry out activities such as movement, growth and development,
and reproduction. But how do living organisms or their cells extract energy from
their environments, and how do cells use this energy to synthesize and
assemble the components from which the cells are made? This module
provides you a conceptual understanding of the biochemical principles of
metabolism so you can figure out how your cells and body utilize various
strategies to meet energy demands.
Module Outcome:
This module enables the students to understand the metabolic processes that
carbohydrates, lipids, and amino acids undergo and integrate the overall design
of metabolism.
Lesson 1
Lesson Title: Carbohydrate Metabolism
Introduction
Welcome to Lesson 1 of module 3! This lesson covers the overview of
metabolism, the carbohydrate catabolic pathways such as glycolysis,
glycogenolysis, Krebs cycle, hexose monophosphate shunt and anabolic
pathways such as gluconeogenesis, biosynthesis of polysaccharides,
and photosynthesis. Electron transport chain and oxidative
phosphorylation are also included in this lesson.
Learning Outcomes
At the end of this lesson, you will:
✓ Distinguish between anabolism and catabolism;
✓ Understand the metabolic process that carbohydrates undergo;
and
✓ Discuss the enzyme, cofactors, regulation, and significance of
carbohydrate metabolism
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ACTIVITY
Laboratory Activity # 2
Fermentation
Objectives:
1. To observe the process of cellular respiration in a living organism, yeast.
2. To perceive how changing food concentrations can affect the rate of
cellular respiration.
Materials Needed:
8 bottles
Active dry yeast
Water
White sugar
8 Balloon
Weighing scale/measuring spoon/measuring cup/graduated cylinder
String
Ruler/tape measure
Thermometer
Procedure:
1) Gather the needed materials. Label the bottles 1, 2, 3, 4. Do also for the
remaining 4 bottles as replicates.
2) Pre-stretch the balloons. Do this by stretching them and blowing them up

and letting out the air.
3) Set up the following bottles using the procedure for each bottle. Make sure
to be accurate in your measurements.
Bottle #1: ½ cup or 118 mL of warm water (40⁰C)

2 tsp or 8.4 grams of sugar
Put the pre-stretched balloon immediately over the top of the flask

1 tsp or 3.1 gram of instant dry yeast

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Bottle #:4 ½ cup or 118 mL of warm water (40⁰C)

4) Gently stir the flask to mix the materials in each flask. Allow the flasks to sit
for 8-12 hours.
5) Use a piece of string to measure the circumference (in cm) of each balloon.
If the balloon does not look noticeably different from the start of the
experiment, calculate the circumference of the balloon as zero.
6) Do the same for the replicates.
7) Fill out the data table.
Data/Result:
Bottle # Contents Circumference of the
balloon (cm)
ANALYSIS
Guide Questions for Discussion:
1) What process was observed in this lab?
2) What gas was produced in this process? What observation was seen to show
this production?
3) What is the overall equation of cellular respiration?
4) Which bottle or bottles had no observable gas production? Why is this so?
5) Which flask produced the largest volume of gas? Why might this be so?
6) What are some observable indicators that a chemical reaction was taking
place inside some of the flasks?
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ABSTRACTION
All living cells require energy to carry out various cellular activities. This energy
is stored in the chemical bonds of organic molecules, such as carbohydrates,
fats, and proteins, that we eat as food. These organic molecules are broken
down by enzymatic reactions in cells to generate energy in the form of
adenosine triphosphate (ATP). The ATP generated by these pathways in cells
is used to drive fundamental cellular processes.
Metabolism is the total of all chemical reactions in an organism. The reaction

pathways that comprise metabolism are divided into two categories.
• Catabolism: degradation of molecules to provide energy.
• Anabolism: reactions using energy to synthesize new molecules for
growth.
Source: https://simplebiologyy.blogspot.com
In anabolic reactions, simple organic molecules such as pyruvic acid, acetyl

unit or intermediate compounds of citric acid cycle serve as starting molecules
for varied biosynthetic products. The energy rich molecules such as ATP or
NADPH derived from catabolic reactions are utilized in the biosynthetic
reactions.
In catabolic reactions, complex substances are broken down to simpler

compounds with a concomitant release of free energy. The released free
energy during these catabolic reactions is conserved in the form of ATP or
NADPH. The major nutrients such as carbohydrates, lipids and proteins are
converted to common intermediate and further metabolised in a central
oxidative pathway.
Stages of Catabolism
1. Stage I - carbohydrates, fats, and proteins are broken down into their
individual monomer units: carbohydrates into simple sugars, fats into
fatty acids and glycerol, and proteins into amino acids. One part of stage
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I of catabolism is the breakdown of food molecules by hydrolysis

reactions into the individual monomer units—which occurs in the mouth,
stomach, and small intestine—and is referred to as digestion.
2. Stage II - the monomer units (or building blocks) are further broken
down through different reaction pathways, one of which produces ATP,
to form a common end product that can then be used in stage III to
produce even more ATP.
3. Stage III - oxidation of the reducing equivalents by oxygen with the

production of ATP.
Source: slideserve.com
Digestion, absorption, and transport of Carbohydrates

Carbohydrate digestion begins in the mouth where salivary α-amylase
attacks the α-glycosidic linkages in starch, the main carbohydrate ingested
by humans. Cleavage of the glycosidic linkages produces a mixture of
dextrins, maltose, and glucose. The α-amylase mixed into the food remains
active as the food passes through the esophagus, but it is rapidly inactivated
in the acidic environment of the stomach. The primary site of carbohydrate
digestion is the small intestine. The secretion of α-amylase in the small
intestine converts any remaining starch molecules, as well as the dextrins,
to maltose. Maltose is then cleaved into two glucose molecules by maltase.
Disaccharides such as sucrose and lactose are not digested until they reach
the small intestine, where they are acted on by sucrase and lactase,
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respectively. The major products of the complete hydrolysis of disaccharides

and polysaccharides are three monosaccharide units: glucose, fructose, and
galactose.
Digestion of Carbohydrate
Source: pressbooks.bccampus.ca
The monosaccharides (glucose, galactose, and fructose) are taken up

through the intestinal epithelial cells. Cotransport of the major
monosaccharide, glucose, and Na+ is driven by a Na+ concentration gradient
that is established by the sodium-potassium pump. Diffusion of Na+ down its
concentration gradient provides the energy to transport glucose across the
cell membrane. This mechanism is also used for galactose transport, while
fructose is taken up by facilitated diffusion. Once inside the intestinal
epithelial cell, monosaccharides are transported into the capillaries of the
intestinal villi and are carried by the hepatic portal system to the liver. Liver
cells convert different types of monosaccharides to glucose, which then
leaves the liver via the circulation to be distributed throughout the body.
Glucose enters the cells by facilitated diffusion. The rate of glucose transport
into most types of cells is greatly influenced by insulin and can increase
tenfold in the presence of insulin. Without insulin, glucose enters most cells
very slowly.
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Transport of glucose across the intestinal epithelium

Source: www.brainkart.com
Carbohydrate Metabolism
Major Pathways in Carbohydrate Metabolism

Source: McKee and McKee, Biochemistry
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Glycolysis
➢ Derived from the Greek words:
Glykys - “sweet” or “sugar”
Lysis - “splitting”
➢ Also referred to as Embden-Meyerhof-Parnas pathway
➢ During glycolysis, a molecule of glucose is degraded in a series of
enzyme-catalyzed reactions to yield two molecules of the three-carbon
compound pyruvate.
➢ During the sequential reactions of glycolysis, some of the free energy
released from glucose is conserved in the form of ATP and NADH.
➢ All the reaction steps take place in the cytoplasm (The enzymes of this
pathway are present in the cytosomal fraction of the cell).
➢ Glycolysis can occur with or without oxygen.
Importance of the glycolytic pathway:

✓ It is the only pathway that is taking place in all the cells of the body.
✓ 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
nonessential amino acids as well as glycerol part of fat.
The glycolytic pathway

The breakdown of the six-carbon glucose into two molecules of the three-
carbon pyruvate occurs in 10 steps and can be divided into two stages:
1. Stage 1 (Preparatory phase)
✓ reactions 1-5
✓ the glucose is phosphorylated, converted to fructose which is
again phosphorylated and cleaved into two molecules of
glyceraldehyde-3-phosphate
✓ In this phase there is an investment of two molecules of ATP.
2. Stage 2 (Energy yielding phase)

✓ the payoff phase
✓ reactions 6-10
✓ the two molecules of glyceraldehyde-3-phosphate are converted
to pyruvate with concomitant generation of four ATP molecules
and two molecules of NADH. Thus, there is a net gain of two ATP
molecules per molecule of Glucose in glycolysis.
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The glycolytic pathway

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Step 1. Phosphorylation of Glucose

Glucose is phosphorylated to glucose-6-phosphate.
The enzyme hexokinase or glucokinase splits ATP into ADP and the Pi
is added on to the glucose producing glucose 6-phosphate, the reaction
is irreversible. Hexokinase is found in muscle and glucokinase in liver.
These enzymes have a requirement for Mg2+ ions, an essential
cofactor. G-6-P inhibits these enzymes (product feedback inhibition),
whereas Pi activates.
Source: Lehninger Principles of Biochemistry
Step 2. Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate

The enzyme phosphohexose isomerase (phosphoglucose isomerase)
catalyzes the reversible isomerization of glucose 6-phosphate, an
aldose, to fructose 6- phosphate, a ketose
Step 3. Phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate

The enzyme phosphofructokinase-1 (PFK-1) uses another ATP
molecule to transfer a phosphate group to fructose -6-phosphate to
form fructose-1, 6-bisphosphate.
The PFK-1 reaction is essentially irreversible under cellular conditions.
PFK-1 is the key regulatory enzyme for glycolysis; it regulates the flux
into pathway and is the first committed step for glycolysis.
Phosphofructokinase-1 is subject to complex allosteric regulation; its
activity is increased whenever the cell’s ATP supply is depleted or when
the ATP breakdown products, ADP and AMP (particularly the latter),
accumulate. The enzyme is inhibited whenever the cell has ample ATP
and is well supplied by other fuels such as fatty acids.
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Step 4. Cleavage of fructose 1,6-bisphosphate

The enzyme fructose 1,6-bisphosphate aldolase, often called simply
aldolase, catalyzes a reversible aldol condensation. Fructose-1, 6-
bisphosphate is cleaved into two 3 carbon atoms; one glyceraldehyde-
3-phosphate (an aldose) and another molecule of dihydroxyacetone
phosphate (a ketose).
Step 5. Interconversion of the Triose Phosphates

Only one of the two triose phosphates formed by aldolase,
glyceraldehyde 3-phosphate, can be directly degraded in the
subsequent steps of glycolysis. The other product, dihydroxyacetone
phosphate, is rapidly and reversibly converted to glyceraldehyde 3-
phosphate by the fifth enzyme of the glycolytic sequence, triose
phosphate isomerase
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Step 6. Oxidation of Glyceraldehyde 3-Phosphate to 1,3-Bisphosphoglycerate

Glyceraldehyde-3-phosphate undergoes two changes simultaneously
under the influence of the enzyme glyceraldehyde-3-phosphate
dehydrogenase. As its name implies, this enzyme takes hydrogen away
from its substrate; it also adds another phosphate group to form 1,3-
bisphosphoglycerate. The hydrogen is removed by the coenzyme
NAD+. For this process to continue, there must be a constant renewal
of the supply of NAD+; this can be achieved in a number of ways
depending upon the presence or absence of oxygen.
Step 7. Phosphoryl Transfer from 1,3-Bisphosphoglycerate to ADP

The enzyme phosphoglycerate kinase transfers the high-energy
phosphoryl group from the carboxyl group of 1,3-bisphosphoglycerate
to ADP, forming ATP and 3-phosphoglycerate
Step 8. Conversion of 3-Phosphoglycerate to 2-Phosphoglycerate

The enzyme phosphoglycerate mutase catalyzes a reversible shift of
the phosphoryl group between C-2 and C-3 of glycerate; Mg2+ is
essential for this reaction
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Step 9. Dehydration of 2-Phosphoglycerate to Phosphoenolpyruvate

The enzyme enolase removes a molecule of water from 2-
phosphoglycerate to form phosphoenolpyruvate (PEP).
Step 10. Transfer of the Phosphoryl Group from Phosphoenolpyruvate to ADP

The last step in glycolysis is the transfer of the phosphoryl group from
phosphoenolpyruvate to ADP, catalyzed by pyruvate kinase, which
requires K+ and either Mg2+ or Mn2+
The entry of other sugars in glycolytic pathway

Although glucose is important, it is not the only sugar that is encountered in
animals and plants. As examples, many plants both produce and use large
quantities of fructose. The suckling mammal obtains half of its sugar from the
galactose of milk lactose.
It is not difficult to see how fructose enters glycolysis, it merely needs to be

phosphorylated to fructose-6-phosphate to enter the pathway. Many plants and
animals have a specific enzyme, fructokinase, which catalyses the reaction:
fructose + ATP fructose-6-phosphate + ADP
In animals, hexokinase can phosphorylate fructose as well as glucose.
The entry of galactose is not as simple as that of fructose: it must be converted

into glucose-6-phosphate before it can be used. During many sugar
transformations the sugar molecule must be attached to a nucleotide. The
nucleotide used in these transformations is uridine diphosphate (UDP).
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Galactose is phosphorylated by galactokinase to give galactose 1-phosphate.
Galactose 1-phosphate uridylyl transferase catalyzes the transfer of

a uridyl group from UDP-glucose to galactose 1-phosphate to form UDP-
galactose and glucose 1-phosphate. The UDP-galactose is converted back to
UDP-glucose by UDP-galactose 4-epimerase. Thus, overall, UDP-glucose is
not consumed in the reaction pathway.
Finally, the glucose 1-phosphate is converted to glucose 6-phosphate by

phosphoglucomutase. The glucose 6-phosphate then enters glycolysis.
Conversion of galactose to glucose 1-phosphate

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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.
Metabolism of other important Sugars

The fate of Pyruvate

In terms of energy, the result of glycolysis is the production of two ATPs and
two NADHs per molecule of glucose.
Pyruvate (other product of glycolysis) is still an energy rich molecule, which
can produce a substantial amount of ATP.
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Under aerobic conditions, the pyruvate formed in the final step of glycolysis
is oxidized to acetate (acetyl CoA), which enters the citric acid cycle and is
oxidized to CO2 and H2O. The NADH formed by dehydrogenation of
glyceraldehyde 3-phosphate is ultimately re-oxidized to NAD+ by passage of
its electrons to O2 in mitochondrial respiration.
Under anaerobic (no oxygen) or hypoxic (low-oxygen) conditions (e.g. in very
active skeletal muscle, in submerged plant tissues, in solid tumors, or in lactic
acid bacteria) NADH generated by glycolysis cannot be re-oxidized by O2.
Failure to regenerate NAD+ would leave the cell with no electron acceptor for
the oxidation of glyceraldehyde 3-phosphate, and the energy-yielding
reactions of glycolysis would stop. NAD+ must therefore be regenerated in
some other mechanism.
The fates of pyruvate

Mechanism 1: Lactic acid fermentation

NADH can be used to reduce pyruvate to lactate to regenerate NAD +.
The reduction of pyruvate in this pathway is catalyzed by lactate
dehydrogenase, which forms the L isomer of lactate at pH 7:
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In glycolysis, dehydrogenation of the two molecules of glyceraldehyde 3-

phosphate derived from each molecule of glucose converts two
molecules of NAD+ to two of NADH. Because the reduction of two
molecules of pyruvate to two of lactate regenerates two molecules of
NAD+, there is no net change in NAD+ or NADH:
Lactic acid fermentation is the basis for lactic acid build up during
vigorous anaerobic muscle contraction: burning sensation felt in active
muscles that tells you to stop overworking the body. Lactate formed in
muscle is eventually carried to the liver by the blood, where it is
converted back into glucose. The cycle of reactions that includes glucose
conversion to lactate in muscle and lactate conversion to glucose in liver
is called the Cori cycle.
Mechanism 2: Ethanol fermentation

Yeast and other microorganisms ferment glucose to ethanol and CO2.
The pyruvate is first decarboxylated in an irreversible reaction
catalyzed by pyruvate decarboxylase yielding acetaldehyde + CO2.
Pyruvate decarboxylase requires Mg2+ and has a tightly bound
coenzyme, thiamine pyrophosphate.
Acetaldehyde is then reduced to ethanol through the action of alcohol
dehydrogenase, with the reducing power furnished by NADH derived
from the dehydrogenation of glyceraldehyde 3-phosphate.
The end products of the combined reactions are CO2 and ethanol
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Recycling of NADH during anaerobic glycolysis

Source: chemistry.gravitywaves.com
Gluconeogenesis
Gluconeogenesis is the synthesis of new glucose molecules from pyruvate,
lactate, glycerol, or the amino acids alanine or glutamine. The vast majority
of gluconeogenesis takes place in the liver and, to a smaller extent, in the
cortex of kidneys. The process occurs during periods of low glucose, that is,
under conditions of fasting, starvation, and low carbohydrate diets.
Brain and red blood cells rely exclusively on glucose as their energy source;
therefore, it is essential that the body maintain a minimum blood glucose
concentration. When the blood glucose concentration falls below that certain
point, new glucose is synthesized by the liver to raise the blood concentration
to normal.
The reaction sequence in gluconeogenesis is largely the reverse of

glycolysis. However, three irreversible enzymes must be bypassed in
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gluconeogenesis vs. glycolysis: Hexokinase (or glucokinase),

Phosphofructokinase, and Pyruvate kinase.
Gluconeogenesis and Glycolysis

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The bypass reactions of gluconeogenesis:
1. Synthesis of PEP
Pyruvate is converted to PEP without using the pyruvate kinase reaction.
The pyruvate is first converted to oxaloacetate, which is in turn converted
to PEP. In the first reaction of this process pyruvate carboxylase adds
carbon dioxide to pyruvate with the expenditure of one ATP equivalent
of energy. Biotin, a carboxyl-group transfer cofactor in animals, is
required by this enzyme.
OAA is then decarboxylated and phosphorylated by PEP carboxykinase

in a reaction driven by the hydrolysis of guanosine triphosphate (GTP)
2. Conversion of fructose-1,6-bisphosphate to fructose-6-phosphate

The irreversible PFK-1-catalyzed reaction in glycolysis is bypassed by
fructose-1,6-bisphosphatase
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3. Formation of glucose from glucose-6-phosphate

Glucose-6-phosphatase (found only in liver and kidney) catalyzes the
irreversible hydrolysis of glucose-6-phosphate to form glucose and Pi.
Glucose is subsequently released into the blood.
Glucose-6-phosphatase
6 CH OPO 2− CH2OH
2 3
5 O O
H H H H
H H2O H
4
OH H 1
OH H + Pi
OH OH OH OH
3 2
H OH H OH
glucose-6-phosphate glucose
Gluconeogenesis substrates:
1. Lactate
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2. Glycerol – product of fat metabolism in adipose tissue
3. Alanine
Pentose Phosphate Pathway

Also known as:
✓ Pentose shunt
✓ Hexose monophosphate shunt
✓ Phosphogluconate pathway
The Pentose Phosphate Pathway is an alternate pathway for glucose oxidation

which is used to provide reducing equivalents in support of biosynthesis. Thus
although it involves the catabolism of glucose, it is generally going to be active
only when anabolism is taking place. All reactions are cytosolic and begins with
the glycolytic intermediate glucose 6-P.
Pentose pathway is often referred to as a shunt because it reconnects with

glycolysis as two of the end products of the pentose pathway are
glyceraldehyde 3-P and fructose 6-P; two intermediates further down in the
glycolytic pathway.
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Glycolysis and Pentose Phosphate Pathway

Main functions of PPP:

1. Generation of NADPH (serves as electron donor)
Pathways requiring NADPH:
✓ Synthesis:
• Fatty acid biosynthesis
• Cholesterol biosynthesis
• Neurotransmitter biosynthesis
• Nucleotide biosynthesis
✓ Detoxification
• Reduction of oxidized glutathione
• Cytochrome P450 monooxygenases
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2. Production of ribose residues for nucleotide and nucleic acid synthesis
Two phases of PPP:

1. Oxidative
2. Nonoxidative (the sugar interconversions portion)
General scheme of the pentose phosphate pathway.

Source: https://vdocuments.mx
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Irreversible oxidative reactions
The oxidative portion of pentose phosphate pathway consists of reactions that

lead to formation of ribulose 5-P, CO2, and 2 molecules of NADPH for each
molecule of G-6-P oxidized.
This portion of the pathway is particularly important in the liver and lactating
mammary glands, which are active in the biosynthesis of fatty acids, in adrenal
cortex, which is active in the NADPH-dependent synthesis of steroids, and in
RBCs, which require NADPH to keep glutathione reduced
The first reaction of the pentose

phosphate pathway is the oxidation of
glucose 6-phosphate by glucose 6-
phosphate dehydrogenase (G6PD) to
form 6-phosphoglucono-δ-lactone, an
intramolecular ester. NADP+ is the
electron acceptor, and the overall
equilibrium lies far in the direction of
NADPH formation.
The lactone is hydrolyzed to the free

acid 6-phosphogluconate by a specific
lactonase, then 6-phosphogluconate
undergoes oxidation and
decarboxylation by 6-
phosphogluconate dehydrogenase to
form the ketopentose ribulose 5-
phosphate; the reaction generates a
second molecule of NADPH.
Phosphopentose isomerase converts
ribulose 5-phosphate to its aldose
isomer, ribose 5-phosphate.
The pentose phosphate pathway is

regulated primarily at the G6PD
reaction. NADPH is a potent
competitive inhibitor of the enzyme,
and, under metabolic conditions, the
ratio of NADPH/NADP+ is sufficiently
high to substantially inhibit enzyme
activity.
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Reversible nonoxidative reactions
The nonoxidative reactions of pentose phosphate pathway occurs in all cell

types synthesizing nucleotides & nucleic acids.
These reactions catalyze the interconversion of 3-, 4-, 5-, 6-, & 7-carbon sugars.
These reversible reactions permit ribulose-5-P (produced by oxidative portion
of pathway) to be converted either to ribose 5-P (needed for nucleotide
synthesis) or to intermediates of glycolysis, F-6-P and glyceraldehyde 3-P.
E.g., many cells that carry out reductive biosynthetic reactions have a greater
need for NADPH than for ribose-5-P. In this case, transketolase (which
transfers 2-C units) and transaldolase (which transfers 3-C units) convert
ribulose 5-P produced as an end-product of the oxidative reactions to
glyceraldehyde 3-P and F-6-P, which are intermediates of glycolysis.
In contrast, under conditions in which the demand for ribose for incorporation
into nucleotides and nucleic acids is greater than the need for NADPH, the
nonoxidative reactions can provide the biosynthesis of ribose-5-P from
glyceraldehyde 3-P & F-6-P in the absence of the oxidative steps
Nonoxidative reactions of the pentose phosphate pathway.

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Role of NADPH in regulating the partitioning of glucose 6-phosphate between

glycolysis and the pentose phosphate pathway.
When NADPH is forming faster than it is being used for biosynthesis and
glutathione reduction, concentration of NADPH rises and inhibits the first
enzyme in the pentose phosphate pathway. As a result, more glucose 6-
phosphate is available for glycolysis.
Glycogen Metabolism
In the liver and muscles, most of the glucose is changed into glycogen by
the process of glycogenesis (anabolism). Glycogen is stored in the liver and
muscles until needed at some later time when glucose levels are low.
If blood glucose levels are low, then epinephrine and glucagon hormones
are secreted to stimulate the conversion of glycogen to glucose. This process
is called glycogenolysis (catabolism).
Glycogenesis
Glycogenesis is the formation of glycogen from glucose. Glycogen is
synthesized depending on the demand for glucose and ATP (energy). If
both are present in relatively high amounts, then the excess of insulin
promotes the glucose conversion into glycogen for storage in liver and
muscle cells.
In the synthesis of glycogen, one ATP is required per glucose incorporated
into the polymeric branched structure of glycogen. Glucose-6-phosphate is
synthesized directly from glucose or as the end product of
gluconeogenesis.
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Steps of glycogenesis
Glycogenolysis
In glycogenolysis, glycogen stored in the liver and muscles, is converted
first to glucose-1- phosphate and then into glucose-6-phosphate. Two
hormones which control glycogenolysis are a peptide, glucagon from the
pancreas and epinephrine from the adrenal glands.
Glucagon is released from the pancreas in response to low blood glucose

and epinephrine is released in response to a threat or stress. Both
hormones act upon enzymes to stimulate glycogen phosphorylase to begin
glycogenolysis and inhibit glycogen synthetase (to stop glycogenesis).
Glycogen is a highly branched polymeric structure containing glucose as

the basic monomer. First individual glucose molecules are hydrolyzed from
the chain, followed by the addition of a phosphate group at C-1. In the next
step the phosphate is moved to the C-6 position to give glucose 6-
phosphate, a cross road compound.
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Glucose-6-phosphate is the first step of the glycolysis pathway if glycogen

is the carbohydrate source and further energy is needed. If energy is not
immediately needed, the glucose-6-phosphate is converted to glucose for
distribution in the blood to various cells such as brain cells.
Steps of glycogenolysis
Cellular Respiration
➢ Recall that glycolysis produces two 3-carbon molecules of pyruvate
from one 6-carbon molecule of glucose; in the process a net of 2
molecules of ATP and 2 molecules of NADH are produced (ATP and
NADH are both high-energy molecules).
➢ When oxygen is present, it is possible to get much more energy out of

glucose by completely oxidizing glucose all the way to carbon dioxide.
➢ The aerobic phase of catabolism, where cells consume O2 and produce

CO2, is called “cellular respiration”.
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There are three stages of cellular respiration:
Stage I - Acetyl-CoA production

Organic fuel molecules
(glucose, fatty acids, and
some amino acids) are
oxidized to yield two-carbon
fragments in the form of the
acetyl group of acetyl-
coenzyme A (acetyl-CoA).
Stage II - Acetyl-CoA oxidation

The acetyl groups are fed into
the citric acid cycle, which
enzymatically oxidizes them to
CO2; the energy released is
conserved in the reduced
electron carriers NADH and
FADH2
Stage III – Electron transfer and

oxidative phosphorylation
The reduced coenzymes
(NADH and FADH2) are
themselves oxidized, giving
up protons (H+) and electrons.
The electrons are transferred
to O2, the final electron
acceptor, via a chain of
electron-carrying molecules
known as the respiratory
chain.
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Conversion of Pyruvate to Acetyl COA

The transition reaction connects glycolysis to the citric acid cycle.
The three-carbon pyruvate molecule generated during glycolysis moves

from the cytoplasm into the mitochondrial matrix, where it is converted by
the enzyme pyruvate dehydrogenase into a two-carbon acetyl coenzyme A
(acetyl CoA) molecule. This reaction is an oxidative decarboxylation
reaction. It converts the three-carbon pyruvate into a two-carbon acetyl CoA
molecule, releasing carbon dioxide and transferring two electrons that
combine with NAD+ to form NADH.
The combined dehydrogenation and decarboxylation of pyruvate to the

acetyl group of acetyl-CoA requires the sequential action of three different
enzymes and five different coenzymes or prosthetic groups.
Pyruvate Dehydrogenase (PDH) complex is composed of multiple copies

of three enzymes:
1. pyruvate dehydrogenase (E1)
2. dihydrolipoyl transacetylase (E2)
3. dihydrolipoyl dehydrogenase (E3)
PDH requires 5 coenzymes or prosthetic group:

• TPP (thiamine pyrophosphate)
• Lipoic Acid (lipoate)
• Coenzyme A
• FAD
• NAD
TCA Cycle
Citric Acid Cycle/Krebs Cycle/Tricarboxylic Acid Cycle
The Tricarboxylic acid cycle is in many ways the central pathway of

metabolism, both catabolically and anabolically: it is involved in the
breakdown and synthesis of a variety of compounds.
The TCA is a cyclic pathway: each product is a substrate for the subsequent
reaction; after a complete turn through the cycle the initial substrate is
regenerated as a product.
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Step 1
The citric acid cycle begins when Coenzyme A transfers its 2-carbon acetyl
group to the 4-carbon compound oxaloacetate to form the 6-carbon
molecule citrate. CoA is subsequently released and can combine with
another pyruvate molecule to begin the cycle again
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Step 2
The citrate is rearranged to form an isomeric form, isocitrate. A hydroxyl
group and a hydrogen molecule are removed from the citrate structure in
the form of water.
The two carbons form a double bond until the water molecule is added
back. Only now, the hydroxyl group and hydrogen molecule are reversed
with respect to the original structure of the citrate molecule. Thus, isocitrate
is formed.
Step 3
The isocitrate molecule is oxidized by a NAD molecule. The NAD molecule
is reduced by the hydrogen atom and the hydroxyl group. The NAD binds
with a hydrogen atom and carries off the other hydrogen atom leaving a
carbonyl group. This structure is very unstable, so a molecule of CO 2 is
released creating alpha-ketoglutarate.
Step 4
In this step, coenzyme A, returns to oxidize the alpha-ketoglutarate
molecule. A molecule of NAD is reduced again to form NADH and leaves
with another hydrogen. This instability causes a carbonyl group to be
released as carbon dioxide and a thioester bond is formed in its place
between the former alpha-ketoglutarate and coenzyme A to create a
molecule of succinyl-coenzyme A complex.
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Step 5
CoA is removed from succinyl-CoA to produce succinate. The energy
released is used to make guanosine triphosphate (GTP) from guanosine
diphosphate (GDP) and Pi by substrate-level phosphorylation. GTP can
then be used to make ATP.
Step 6
Succinate is oxidized by a molecule of FAD (Flavin adenine dinucleotide).
The FAD removes two hydrogen atoms from the succinate and forces a
double bond to form between the two carbon atoms, thus creating fumarate.
Step 7
An enzyme adds water to the fumarate molecule to form malate. The
malate is created by adding one hydrogen atom to a carbon atom and then
adding a hydroxyl group to a carbon next to a terminal carbonyl group.
Step 8
The malate molecule is oxidized by a NAD molecule. The carbon that
carried the hydroxyl group is now converted into a carbonyl group. The end
product is oxaloacetate which can then combine with acetyl-coenzyme A
and begin the Krebs cycle all over again.
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Oxaloacetate is then ready to combine with the next acetyl CoA to start the
Krebs cycle again. For each turn of the cycle, three NADH, one ATP
(through GTP), and one FADH2 are created. Each carbon of pyruvate is
converted into CO2, which is released as a by-product of oxidative (aerobic)
respiration.
The NADH + H+ and FADH2 carry protons and electrons to the electron
transport chain to generate additional ATP by oxidative phosphorylation
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Source: https://www.slideserve.com
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Impermeable to ions and

most other compounds
Electron Transport Chain (ETC)

Electrons carried by reduced coenzymes (NADH or FADH2) are passed
sequentially through a chain of proteins and coenzymes (so called electron
transport chain) to O2.
The electron transport chain is a series of protein complexes and electron

carrier molecules within the inner membrane of mitochondria that generate ATP
for energy.
Electrons are passed along the chain from protein complex to protein complex
until they are donated to oxygen. During the passage of electrons, protons are
pumped out of the mitochondrial matrix across the inner membrane and into
the intermembrane space.
The accumulation of protons in the intermembrane space creates an

electrochemical gradient that causes protons to flow down the gradient and
back into the matrix through ATP synthase. This movement of protons provides
the energy to produce ATP.
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Protein Complexes in the Electron Transport Chain

There are four protein complexes that are part of the electron transport chain
that functions to pass electrons down the chain. These complexes are
embedded within the inner mitochondrial membrane.
Complex I or NADH dehydrogenase or NADH:ubiquinone oxidoreductase

NADH transfers two electrons to Complex I resulting in four H+ ions being
pumped across the inner membrane. NADH is oxidized to NAD+, which is
recycled back into the Krebs cycle. Electrons are transferred from Complex
I to a carrier molecule ubiquinone (Q), which is reduced to ubiquinol (QH2).
Ubiquinol carries the electrons to Complex III.
Complex II or Succinate dehydrogenase

FADH2 transfers electrons to Complex II and the electrons are passed
along to ubiquinone (Q). Q is reduced to ubiquinol (QH2), which carries the
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electrons to Complex III. No H+ ions are transported to the intermembrane

space in this process.
Complex III or cytochrome bc1 complex or ubiquinone:cytochrome c oxidore-

ductase
The passage of electrons to Complex III drives the transport of four more
H+ ions across the inner membrane. QH2 is oxidized and electrons are
passed to another electron carrier protein cytochrome C.
Complex IV - also called cytochrome oxidase

Cytochrome C passes electrons to the final protein complex in the chain,
Complex IV. Two H+ ions are pumped across the inner membrane. The
electrons are then passed from Complex IV to an oxygen (O 2) molecule,
causing the molecule to split. The resulting oxygen atoms quickly grab H+
ions to form two molecules of water.
ATP Synthase
ATP synthase moves H+ ions that were pumped out of the matrix by the
electron transport chain back into the matrix. The energy from the influx
of protons into the matrix is used to generate ATP by the phosphorylation
(addition of a phosphate) of ADP. The movement of ions across the
selectively permeable mitochondrial membrane and down their
electrochemical gradient is called chemiosmosis. (The production of ATP
using the process of chemiosmosis in mitochondria is called oxidative
phosphorylation).
If the membrane were open to diffusion by the hydrogen ions, the ions
would tend to diffuse back across into the matrix, driven by their
electrochemical gradient. Recall that many ions cannot diffuse through the
nonpolar regions of phospholipid membranes without the aid of ion
channels. Similarly, hydrogen ions in the matrix space can only pass
through the inner mitochondrial membrane through an integral membrane
protein called ATP synthase. This complex protein acts as a tiny
generator, turned by the force of the hydrogen ions diffusing through it,
down their electrochemical gradient. The turning of parts of this molecular
machine facilitates the addition of a phosphate to ADP, forming ATP,
using the potential energy of the hydrogen ion gradient.
NADH generates more ATP than FADH2. For every NADH molecule that
is oxidized, 10 H+ ions are pumped into the intermembrane space. This
yields about 2.5 ATP molecules. Because FADH2 enters the chain at a
later stage (Complex II), only six H+ ions are transferred to the
intermembrane space. This accounts for about 1.5 ATP molecules. A total
of 30-32 ATP molecules are generated in electron transport and oxidative
phosphorylation.
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Shuttle Systems Indirectly Convey Cytosolic NADH into Mitochondria for

Oxidation
The NADH dehydrogenase of the inner mitochondrial membrane of animal
cells can accept electrons only from NADH in the matrix. Given that the inner
membrane is not permeable to NADH, how can the NADH generated by
glycolysis in the cytosol be reoxidized to NAD by O2 via the respiratory
chain?
Special shuttle systems carry reducing equivalents from cytosolic NADH

into mitochondria by an indirect route. The most active NADH shuttle,
which functions in liver, kidney, and heart mitochondria, is the malate-
aspartate shuttle.
Malate-Aspartate Shuttle
The reducing equivalents of cytosolic NADH are first transferred to

cytosolic oxaloacetate to yield malate, catalyzed by cytosolic malate
dehydrogenase. The malate thus formed passes through the inner
membrane via the malate-ketoglutarate transporter. Within the matrix the
reducing equivalents are passed to NAD by the action of matrix malate
dehydrogenase, forming NADH; this NADH can pass electrons directly
to the respiratory chain. About 2.5 molecules of ATP are generated as
this pair of electrons passes to O2.
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Cytosolic oxaloacetate must be regenerated by transamination reactions

and the activity of membrane transporters to start another cycle of the
shuttle.
Skeletal muscle and brain use a different NADH shuttle, the glycerol 3-
phosphate shuttle. It differs from the malate-aspartate shuttle in that it
delivers the reducing equivalents from NADH to ubiquinone and thus into
Complex III, not Complex I, providing only enough energy to synthesize
1.5 ATP molecules per pair of electrons.
The mitochondria of plants have an externally oriented NADH

dehydrogenase that can transfer electrons directly from cytosolic NADH
into the respiratory chain at the level of ubiquinone. Because this
pathway bypasses the NADH dehydrogenase of Complex I and the
associated proton movement, the yield of ATP from cytosolic NADH is
less than that from NADH generated in the matrix.
Glycerol-3-phosphate shuttle
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ATP yield from complete oxidation of glucose
Photosynthesis
Source: https://es.vecteezy.com
Photosynthesis is the process in which the energy of light is used to bring about
synthesis of complex organic molecules, such as sugars, from carbon dioxide
and water. It is of enormous importance as it is the principal means by which
energy can enter biological systems.
Photosynthesis occurs in a wide range of organisms, from green plants and

algae to some bacteria. The processes themselves are always associated with
biological membranes in which pigments are embedded. In eukaryotes such as
green plants and green algae, these membranes are found in organelles called
chloroplasts, but in prokaryotes such as photosynthetic bacteria and blue-green
algae, the pigments are present in specialized membranes within the
cytoplasm.
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Actual site for photosynthesis in plants
Importance of Photosynthesis
✓ Photosynthesis is essential for the existence of all life on earth. It serves a
crucial role in the food chain – the plants create their food using this
process, thereby, forming the primary producers.
✓ Photosynthesis is also responsible for the production of oxygen – which is
needed by most organisms for their survival.
The process of photosynthesis can be divided into two parts.

1. Light reactions
✓ convert solar energy to chemical energy
✓ produce ATP and NADPH
✓ the light reactions are brought about by the flow the flow of
electrons through two photosystems: photosystem I (PS I) and
photosystem II (PS II). This electron flow generates a proton
gradient across the membranes and this in turn drives the
synthesis of ATP by an enzyme called ATP synthase
2. Dark reactions or light-independent reactions

✓ ATP and NADPH generated by the light reactions are used to
reduce carbon dioxide to produce sugars
✓ these reactions are brought about by the operation of a metabolic
pathway usually known as the Calvin cycle.
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Two sets and reactions during photosynthesis.

Source: https://www.slideserve.com
Photosynthetic pigments
Pigment molecules are responsible for absorbing light and form part of
discrete structural components of the membranes called photosystems,
which are embedded in the thylakoid membranes.
1. Chlorophylls
The chlorophylls are the main light-absorbing pigments. They
contain a magnesium atom chelated by the pyrrole nitrogen atoms
of a porphyrin ring.
Chlorophyll b differs from chlorophyll a only in having a -CHO group

in ring II where chlorophyll a has a CH3.
Because of the long series of conjugated double bonds (i.e.
alternating double and single bonds) in the rings, chlorophylls absorb
visible light very strongly indeed, Chlorophyll a and chlorophyll b
absorb blue and red light particularly strongly. However, their
absorption spectra do differ slightly from one another so that they
complement each other and allow a wider range of wavelengths of
light to be absorbed
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Absorption spectra of photosynthetic pigments.
Chlorophyll looks green because it absorbs red and blue light, making these
colours unavailable to be seen by our eyes. It is the green light which is NOT
absorbed that finally reaches our eyes, making chlorophyll appear green.
However, it is the energy from the red and blue light that are absorbed that is,
thereby, able to be used to do photosynthesis. The green light we can see is
not/ cannot be absorbed by the plant, and thus cannot be used to do
photosynthesis.
Interaction of light with chloroplasts
2. Carotenoids
The carotenoids are polyisoprenoid compounds with long series of
conjugated double bonds
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There are two classes of carotenoids:

a. carotenes such as β-carotene which contain no oxygen
b. xanthophylls such as lutein which contain oxygen
Carotenoids absorb light of wavelengths between about 400 and

500 nm and therefore have absorption spectra which are
complementary to the chlorophylls and help to trap more light. The
carotenoids are also important antioxidants, preventing photo-
oxidation of chlorophyll at high light intensity.
Pigment Molecules used in photosynthesis

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Factors Affecting Photosynthesis
✓ Light Intensity: Increased light intensity results in a higher rate of

photosynthesis. On the other hand, low light intensity results in a lower
rate of photosynthesis.
✓ The concentration of CO2: Higher concentration of carbon dioxide helps

in increasing the rate of photosynthesis. Usually, carbon dioxide in the
range of 300 – 400 PPM is adequate for photosynthesis.
✓ Temperature: For efficient execution of photosynthesis, it is important to

have a temperature range between 25° to 35° C.
✓ Water: As water is an important factor in photosynthesis, its deficiency

can lead to problems in the intake of carbon dioxide. The scarcity of
water leads to the refusal of stomatal opening to retain the amount of
water they have stored inside.
✓ Pollution: Industrial pollutants and other particulates may settle on the

leaf surface. This can block the pores of stomata which makes it difficult
to take in carbon dioxide.
APPLICATION
1. How are photosynthesis and cellular respiration connected in a

biochemical pathway?
2. Explain the production of 30-32 ATP in the complete oxidation of 1

molecule of glucose.
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Lesson 2
Lesson Title: Lipid Metabolism
Introduction
Welcome to lesson 2 of module 3. This lesson covers lipid metabolism
specifically lipolysis, β-oxidation, ketogenesis, and lipogenesis.
Learning Outcomes
✓ Elucidate the digestion, absorption, and transport of lipids
✓ Describe the anabolic and catabolic pathways of lipids
ACTIVITY
Examine the given foods. Which among them had the highest fat content?
banana bacon grilled chicken breast
ANALYSIS
Which among the foods given in the activity will give your body the highest
energy per unit weight? Expound your answer.
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ABSTRACTION
Digestion, Mobilization, and Transport

Cells can obtain fatty acid fuels from three sources:
• fats consumed in the diet
• fats stored in cells as lipid droplets
• fats synthesized in one organ for export to another
Some species use all three sources under various circumstances, others use
one or two.
Example: Vertebrates obtain fats in the diet, mobilize fats stored in
specialized tissue (adipose tissue, consisting of cells called
adipocytes), and, in the liver, convert excess dietary
carbohydrates to fats for export to other tissues.
Triacylglycerols provide more than half the energy requirements of some

organs, particularly the liver, heart, and resting skeletal muscle.
Stored triacylglycerols are virtually the sole source of energy in hibernating

animals and migrating birds.
Protists obtain fats by consuming organisms lower in the food chain, and
some also store fats as cytosolic lipid droplets.
Vascular plants mobilize fats stored in seeds during germination, but do not
otherwise depend on fats for energy.
Processing of dietary lipids in vertebrates

Lipolysis
To obtain energy from fat, triglycerides must first be broken down by
hydrolysis into their two principal components, fatty acids and glycerol.
This process, called lipolysis, takes place in the cytoplasm. The
resulting fatty acids are oxidized by β-oxidation into acetyl CoA, which
is used by the Krebs cycle. The glycerol that is released from
triglycerides after lipolysis directly enters the glycolysis pathway as
DHAP.
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Entry of glycerol into the glycolytic pathway.

(source: Lehninger Principles of Biochemistry)
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Dietary fat must first be emulsified to increase its surface area for contact
with the water-soluble lipases. This occurs largely in the duodenum after
mixing with the bile acids, a family of cholesterol derived detergents.
Triacylglycerols can then be hydrolyzed by pancreatic lipase to free fatty
acids and 2-monoacylglycerol:
The fatty acids and monoacylglycerol are absorbed by the intestinal cells,
converted to fatty acyl CoA and reassembled into triacylglycerols. The
triacylglycerols then assemble with phospholipids and lipoproteins to form
chylomicrons for transport through the lymph and blood to the tissues.
When the chylomicrons reach tissue cells the triacylglycerols are again
hydrolysed by lipoprotein lipase to fatty acids which can be taken up by
the peripheral tissue cells. In adipose cells the fatty acids are then
converted into fatty acyl CoA's and combined into triacylglycerols for
storage. Alternatively, the fatty acids can be broken down for energy using
the various oxidative pathway in specific tissues.
Processing of dietary lipids in vertebrates

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Mobilization of triacylglycerols stored in adipose tissue
When low levels of glucose in the blood trigger the release of glucagon, (1) the
hormone binds its receptor in the adipocyte membrane and thus (2) stimulates
adenylyl cyclase, via a G protein, to produce cAMP. This activates PKA, which
phosphorylates (3) the hormone-sensitive lipase (HSL) and (4) perilipin
molecules on the surface of the lipid droplet. Phosphorylation of perilipin causes
(5) dissociation of the protein CGI from perilipin. CGI then associates with the
enzyme adipose triacylglycerol lipase (ATGL), activating it. Active ATGL (6)
converts triacylglycerols to diacylglycerols. The phosphorylated perilipin
associates with phosphorylated HSL, allowing access to the surface of the lipid
droplet, where (7) it converts diacylglycerols to monoacylglycerols. A third
lipase, monoacylglycerol lipase (MGL) (8), hydrolyzes monoacylglycerols. (9)
Fatty acids leave the adipocyte, bind serum albumin in the blood, and are
carried in the blood; they are released from the albumin and (10) enter a
myocyte via a specific fatty acid transporter. (11) In the myocyte, fatty acids are
oxidized to CO2, and the energy of oxidation is conserved in ATP, which fuels
muscle contraction and other energy-requiring metabolism in the myocyte.
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Triacylglycerol Breakdown in Plants

In some plants, large quantities of triacylglycerol are stored in seeds or
fruits. In dry seeds prior to germination there is little enzymic activity,
however during imbibition (the uptake of water by seeds during
germination) there is an increase in the activity of several enzymes
including triacylglycerol lipases. Their substrates are contained in oil
droplets (oleosomes) within the seed and the enzymes act at the surface
of the droplet, probably with the help of binding proteins to facilitate the
attachment process. The lipases catalyse the release of fatty acids
esterified to the 1 and 3 position of triacylglycerols, to yield a
monoacylglycerol. Complete release of all three fatty acids is achieved by
the migration of fatty acid from the 2 position to the 1 position and its
subsequent release. The fatty acids released are taken up by the
glyoxysomes where they are oxidized. The mechanism of transfer of fatty
acids between the sites of storage and of oxidation is poorly understood,
but in some seeds, this may be achieved by direct contact between the oil
droplet and the glyoxysome.
The sequential breakdown of triacylglycerols

(source: Chesworth et al., 1998)
Stages of fatty acid oxidation:

Stage 1: A long-chain fatty acid is oxidized to yield acetyl residues in the form
of acetyl-CoA. This process is called β oxidation.
Stage 2: The acetyl groups are oxidized to CO2 via the citric acid cycle.
Stage 3: Electrons derived from the oxidations of stages 1 and 2 pass to O 2

via the mitochondrial respiratory chain, providing the energy for ATP
synthesis by oxidative phosphorylation.
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Stages of fatty acid oxidation

β Oxidation
Most tissues can oxidize fatty acids by β-oxidation. Again, there are two main
sites of β-oxidation in the cell. In animals, the mitochondrial matrix is the site
where fatty acids are completely oxidized to acetyl-CoA. Peroxisomal
oxidation of fatty acids is important in the liver and kidney. β-oxidation in
peroxisomes appears to be a mechanism for shortening the chain length of
fatty acids, which may then be further oxidized in the mitochondrial matrix.
In plant leaf tissue, peroxisomes rather than mitochondria are the main sites
of fatty acid oxidation, whereas in seeds, glyoxysomes are the major sites of
fatty acid oxidation where the product of oxidation, acetyl-CoA, can feed
directly into the glyoxylate pathway.
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In the cytoplasm, fatty acids must be activated to their CoA ester derivatives
before they can be further metabolized. The reaction requires ATP and is
catalysed by enzymes called acyl-CoA synthetases (pls see figure below).
There are a number of different enzymes all of which catalyse the same
general reaction but which have differing chain-length specificities. The two
most important in fatty acid oxidation are:
• medium-chain acyl-CoA synthetase (C4-C12)

• long-chain acyl-CoA synthetase (C10+)
Long-chain acyl-CoA synthetase is a membrane-bound enzyme found on the

peroxisome membrane, the endoplasmic reticulum and the outer
mitochondrial membrane. This distribution allows fatty acids to be
metabolized by various pathways at different subcellular sites, i.e. chain
shortening in peroxisomes, phospholipid and triacylglycerol synthesis on the
endoplasmic reticulum, and β-oxidation in the mitochondria.
The inner mitochondrial membrane presents a barrier to fatty acyl-CoAs

destined for oxidation. Neither CoA itself nor esters of CoA can pass through
this membrane. This is largely due to the large size and charged nature of
the CoA molecule. To facilitate the movement of fatty acids across the inner
mitochondrial membrane to the mitochondrial matrix, acyl-CoAs are
converted to acyl-carnitines by the enzyme carnitine acyltransferase I
located in the mitochondrial intermembrane space. Fatty acyl-carnitine
derivatives cross the inner membrane in exchange for carnitine by means of
a transport protein, acyl-carnitine translocase. In the mitochondrial matrix
fatty acyl-carnitines are converted back to CoA derivatives by carnitine
acyltransferase II. Once in the mitochondrial matrix as CoA esters, fatty acids
can undergo β-oxidation.
Activation and transport of fatty acids to the mitchondrial matrix.
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THE REACTIONS OF β-OXIDATION

The β-oxidation pathway uses a sequence of four reactions to remove a
two-carbon unit from the carboxyl end of the fatty acid. This reaction
sequence is repeated until the fatty acid is completely oxidized.
1. Fatty acyl-CoA is acted upon by an enzyme acyl-CoA dehydrogenase

which is FAD dependent. Fatty acyl-CoA undergoes dehydrogenation
and forms a trans-double bond at the α and β carbons to form trans-Δ2-
enoyl-CoA. Acyl-CoA dehydrogenase are present as three isoenzymes
each specific for a particular carbon chain length (short, intermediate
and long). The electrons which were removed from the fatty acyl-CoA
chain are transferred to FAD which gets reduced to FADH2. This FADH2
immediately via the Electron Transport System gets converted to ATP
molecules.
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2. Enoyl-CoA hydratase catalyses this reaction where water is added.

Hydration occurs at the double bond resulting in the formation of β-
hydroxyacyl-CoA.
3. β-hydroxyacyl-CoA undergoes dehydrogenation to form β-ketoacyl-

CoA in the presence of β-hydroxyacyl-CoA dehydrogenase. The
electrons available as a result of dehydrogenation are accepted by
NAD+ to form NADH + H+ which immediately exchanges these electrons
with oxygen in the Electron Transport System to form ATP molecules.
4. The fourth reaction is called as thiolysis as acyl-CoA acetyltransferase

(also known as thiolase) in the presence of CoA-SH causes the
cleavage of β-ketoacyl-CoA to form acetyl CoA and the thioester of the
original fatty acid with two carbons less. This cleavage occurs as the β
carbon ketone group is a good target for nucleophilic attack by the thiol
(-SH) group of the coenzyme A.
The new fatty acyl-CoA (n-2 carbons) formed again participates in the β-
oxidation cycle to form a new fatty acyl-CoA with two carbons less (n-4
carbons) and a new molecule of acetyl CoA. This process continues till the
entire fatty acid is converted into acetyl CoA molecules.
Acetyl CoA formed from the above steps now enters the Kreb’s cycle to get
oxidized to CO2 and H2O.
Example: Beta-oxidation of saturated 16 C Fatty Acid (palmitate)
✓ 7 rounds of Beta oxidation (bottom numbers)

✓ 8 acetyl CoA formed (top numbers)
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(a) In each pass through the four-step beta oxidation sequence, one acetyl
residue (shaded in pink) is removed in the form of acetyl-CoA from the
carboxyl end of the fatty acyl chain (palmitate) which enters as
palmitoyl-CoA
(b) Six more passes through the pathway yield seven more molecules of
acetylCoA, the seventh arising from the last two carbon atoms of the
16-carbon chain. Eight molecules of acetyl-CoA are formed in all.
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Yield of ATP during Oxidation of One Molecule of palmitate to CO2 and H2O:
*One molecule of ATP is hydrolysed to AMP and PPi during the activation of
a fatty acid prior to transport into the mitochondrial matrix. This is equivalent
to the utilization of 2 ATP.
.
Ketone Bodies
In humans and most other mammals, acetyl-CoA formed in the liver during
oxidation of fatty acids can either enter the citric acid cycle or undergo
conversion to the “ketone bodies,” acetone, acetoacetate, and D-β-
hydroxybutyrate, for export to other tissues.
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Note that only two of the ketone bodies are in fact ketones, and that acetone
is an "unintentional" breakdown product resulting from the instability of
acetoacetate at body temperature. Acetone is not available as fuel to any
significant extent, and is thus a waste product. It is excreted in urine or
exhaled.
Overproduction of the ketone bodies acetoacetate and β-hydroxybutyrate

can lead to ketoacidosis, a serious complication in which excess ketone
bodies (acids) lower the blood’s pH. Slightly raised levels of ketone bodies
is referred to as ketosis.
Ketogenesis (Ketone Synthesis)

✓ If excessive acetyl CoA is created from the oxidation of fatty acids and
the Krebs cycle is overloaded and cannot handle it, the acetyl CoA is
diverted to create ketone bodies.
✓ These ketone bodies can serve as a fuel source if glucose levels are too
low in the body.
✓ Ketones serve as fuel in times of prolonged starvation or when patients
suffer from uncontrolled diabetes and cannot utilize most of the
circulating glucose.
Formation of ketone bodies from acetyl-CoA

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Ketone Body Oxidation

When glucose is limited, ketone bodies can be oxidized to produce acetyl
CoA to be used in the Krebs cycle to generate energy.
D-β-Hydroxybutyrate, synthesized in the liver, passes into the blood and thus
to other tissues, where it is converted in three steps to acetyl-CoA. It is first
oxidized to acetoacetate, which is activated with coenzyme A donated from
succinyl-CoA, then split by thiolase. The acetyl-CoA thus formed is used for
energy production.
Fatty Acid Synthesis Occurs in the Cytosol of Many Organisms but in the
Chloroplasts of Plants
In most higher eukaryotes, the fatty acid synthase complex is found
exclusively in the cytosol. This location segregates synthetic processes
from degradative reactions, many of which take place in the mitochondrial
matrix. There is a corresponding segregation of the electron-carrying
cofactors used in anabolism (generally a reductive process) and those
used in catabolism (generally oxidative).
Usually, NADPH is the electron carrier for anabolic reactions, and NAD
serves in catabolic reactions. In hepatocytes, the [NADPH]/[NADP] ratio
is very high (about 75) in the cytosol, furnishing a strongly reducing
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environment for the reductive synthesis of fatty acids and other

biomolecules. The cytosolic [NADH]/[NAD] ratio is much smaller (only
about 8 x 10-4), so the NAD-dependent oxidative catabolism of glucose
can take place in the same compartment, and at the same time, as fatty
acid synthesis. The [NADH]/[NAD] ratio in the mitochondrion is much
higher than in the cytosol, because of the flow of electrons to NAD from
the oxidation of fatty acids, amino acids, pyruvate, and acetylCoA. This
high mitochondrial [NADH]/[NAD] ratio favors the reduction of oxygen via
the respiratory chain.
In the photosynthetic cells of plants, fatty acid synthesis occurs not in the
cytosol but in the chloroplast stroma. This makes sense, given that
NADPH is produced in chloroplasts by the light reactions of
photosynthesis.
Take note that the resulting high [NADPH]/[NADP] ratio provides the
reducing environment that favors reductive anabolic processes such as
fatty acid synthesis.
Subcellular localization of lipid metabolism

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Lipogenesis
✓ When glucose levels are plentiful, the excess acetyl CoA generated by
glycolysis can be converted into fatty acids, triglycerides, cholesterol,
steroids, and bile salts.
✓ This process, called lipogenesis, creates lipids (fat) from the acetyl CoA and
takes place in the cytoplasm of adipocytes (fat cells) and hepatocytes (liver
cells).
✓ When you eat more glucose or carbohydrates than your body needs, your
system uses acetyl CoA to turn the excess into fat.
Triacylglycerol Synthesis
Glycerol-3-phosphate or DHAP reacts sequentially with three molecules of
acyl-CoA (fatty acid esters of CoASH). In the synthesis of triacylglycerols,
phosphatidic acid is formed by two sequential acylations of glycerol-3-
phosphate or by a pathway involving the direct acylation of DHAP. In the
latter pathway, acyldihydroxyacetone phosphate is later reduced to form
lysophosphatidic acid. Depending on the pathway used, lysophosphatidic
acid synthesis utilizes either an NADH or an NADPH cofactor. Phosphatidic
acid is produced when lysophosphatidic acid reacts with a second acyl-CoA.
Once formed, phosphatidic acid is converted to diacylglycerol by
phosphatidic acid phosphatase. A third acylation reaction forms
triacylglycerol.
Glyceroneogenesis in Adipocytes
Glyceroneogenesis is a major source of glycerol-3-phosphate required for
Triacylglycerol synthesis. Substrates for this pathway include lactate,
pyruvate, and glucogenic amino acids such as alanine. Pyruvate is
converted to OAA within the mitochondrion by PC (pyruvate carboxylase).
After OAA is reduced by NADH, the product, malate, is transported out of
the mitochondrion where the reaction is reversed to form OAA. OAA is then
phosphorylated and decarboxylated by phosphoenolpyruvate carboxykinase
(PEPCK-C) in a GTP-requiring reaction to form phosphoenolpyruvate (PEP).
PEP is then converted via gluconeogenesis to DHAP. DHAP is reduced by
glycerol-3-phosphate dehydrogenase to glycerol-3-phosphate, which is then
utilized in TG synthesis.
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Triacylglycerol Synthesis
(Source: McKee and McKee, Biochemistry)
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Glyceroneogenesis in Adipocytes
(Source: McKee and McKee, Biochemistry)
APPLICATION
How many ATPs are produced in the oxidation of stearic acid?
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Lesson 3
Lesson Title: Nitrogen Metabolism
Introduction
All the living organisms are basically composed of carbon, hydrogen,
oxygen, nitrogen, and many other forms of chemical elements. Nitrogen
is next to carbon in importance in living organisms as it is an important
constituent of amino acids, proteins, enzymes, vitamins, alkaloids, and
some growth hormones.
This lesson covers the nitrogen fixation, breakdown of protein,

degradation, and synthesis of amino acids.
Learning Outcomes
✓ Describe the reactions involved in the degradation of amino
acids
✓ Describe the fate of ammonia in the urea cycle
✓ Designate the fate of carbon skeleton
ACTIVITY
Examine the figure below and answer the questions in the analysis.
182 | P a g e
ANALYSIS
Select the correct answer.
1. Nitrogen fixation is the conversion of

a. N2 to N
b. N2 to NH3
c. N2 to NO3–
d. N2 to urea
2. Important enzymes involved in nitrogen fixation are

a. Nitrogenase and hydrogenase
b. Nitrogenase and hexokinase
c. Nitrogenase and peptidase
d. Nitrogenase and hydrolyase
3. Conversion of nitrates to nitrogen is called

a. Ammonification
b. Nitrification
c. Nitrogen fixation
d. Denitrification
4. Conversion of ammonia to nitrite and then to nitrates is called

a. Ammonification
b. Denitrification
c. Assimilation
d. Nitrification
5. Nitrogen content of biosphere remains constant because of:

a. Nitrogen cycle
b. Nitrogen fixation
c. Industrial pollution
d. Absorption of nitrogen
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ABSTRACTION
Nitrogen Cycle
Nitrogen is primarily present in the atmosphere freely as dinitrogen or nitrogen
gas. Air has 78% N2 but most of the living organisms cannot utilize this
atmospheric Nitrogen as molecular nitrogen or diatomic nitrogen (N2) is highly
stable as it is triple bonded (N≡N). Because of this stability, molecular nitrogen
as such is not very reactive in the atmosphere under normal conditions.
Nitrogen cycle converts this nitrogen into a usable form. Lightning fixes
nitrogen to NH3, and nitrogen fixing bacteria like Rhizobium (which live in roots
of leguminous plants like pea, rajma, beans, pulses etc.) also convert N2 into
NH3. Most plants absorb nitrates from soil and reduce it to NH3 in the cells for
further metabolic reactions. Dead organisms and their excreta like urea are
decomposed by bacteria into NH3 and by a different set of bacteria into
nitrates. These are left in the soil for use by plants. In this way Nitrogen cycle
is self-regulated but human activities have caused steady loss of soil nitrogen.
184 | P a g e
Nitrogen Fixation
The conversion of molecular nitrogen into compounds of nitrogen especially
ammonia is called nitrogen fixation. Nitrogen fixation is a reductive process
i.e., nitrogen fixation will stop if there is no reducing condition or if oxygen is
present.
two different methods of nitrogen fixation:

1. Abiological nitrogen fixation
✓ the nitrogen is reduced to ammonia without involving any living
cell
✓ two types:
a. industrial
For example, in the Haber’s process, synthetic ammonia is
produced by passing a mixture of nitrogen and hydrogen
through a bed of catalyst (iron oxides) at a very high
temperature and pressure.
b. natural
Nitrogen can be fixed especially during electrical discharges
in the atmosphere. It may occur during lightning storms
when nitrogen in the atmosphere can combine with oxygen
to form oxides of nitrogen
These oxides of nitrogen may be hydrated and trickle down

to earth as combined nitrite and nitrate.
2. Biological nitrogen fixation

Chemically, this process is same as abiological. Biological
nitrogen fixation is reduction of molecular nitrogen to ammonia by
a living cell in the presence of enzymes called nitrogenases.
Nitrogen fixation is a distinctive property possessed by a select

group of organisms, because of the presence of the enzyme
nitrogenase in them. The process of nitrogen fixation is primarily
confined to microbial cells like bacteria and cyanobacteria. These
microorganisms may be independent and free living.
185 | P a g e
Examples of organisms which fix nitrogen
Some microbes may become associated with other organisms and fix
nitrogen. The host organism may be a lower plant or higher plant. The
host organism and the nitrogen fixing microbes establish a special
relationship called symbiosis and this results in symbiotic nitrogen
fixation.
Some symbiotic nitrogen fixing organisms
Mechanism of Biological Fixation of Nitrogen

Biological nitrogen fixation can be represented by the following equation, in
which two moles of ammonia are produced from one mole of nitrogen gas,
at the expense of 16 moles of ATP and a supply of electrons and protons
(hydrogen ions):
N2 + 8H+ + 8e- + 16 ATP 2NH3 + H2 + 16 ADP + 16Pi
Enough cell energy in the form of ATP is required for stepwise reduction of
nitrogen to ammonia since molecular nitrogen is a very stable molecule. The
reaction is performed exclusively by prokaryotes (the bacteria and related
186 | P a g e
organisms), using an enzyme complex termed nitrogenase. This enzyme

consists of two proteins - an iron protein and a molybdenum-iron protein.
The reactions occur while N2 is bound to the nitrogenase enzyme complex.

The Fe protein is first reduced by electrons donated by ferredoxin. Then the
reduced Fe protein binds ATP and reduces the molybdenum-iron protein,
which donates electrons to N2, producing HN=NH. In two further cycles of
this process (each requiring electrons donated by ferredoxin) HN=NH is
reduced to H2N-NH2, and this in turn is reduced to 2NH3.
Depending on the type of microorganism, the reduced ferredoxin which

supplies electrons for this process is generated by photosynthesis, respiration,
or fermentation.
In legumes, nitrogen fixation occurs in specialized bodies called root nodules.

The nodules develop due to interaction between the bacteria Rhizobium and
the legume roots.
187 | P a g e
The biochemical steps for nitrogen fixation are same. However, legume
nodules possess special protein called LEGHEMOGLOBIN. The synthesis of
leghemoglobin is the result of symbiosis because neither bacterium alone nor
legume plant alone possess the protein. In addition to leghemoglobin, a group
of proteins called nodulins are also synthesized which help in establishing
symbiosis and maintaining nodule functioning.
Leghemoglobin is considered to lower down the partial pressure of oxygen and

helps in nitrogen fixation. However, this function is specific for legumes only
because free living microbes do not possess nitrogen fixing leghemoglobin.
Moreover, it has also not been found in cyanobacterial symbiosis with other
plants, which fix N2 under aerobic condition.
Nitrate and Ammonia Assimilation by Plants

As mentioned, nitrogen fixation is confined to selected microbes and plants. But
all plants require nitrogen because it has a role to play in the general
metabolism. Therefore, plants which do not fix nitrogen, use other combined
nitrogen sources such as nitrate and ammonia for carrying on metabolic activity.
Nitrate is absorbed by most plants and reduced to ammonia with the help of
two different enzymes. The first step conversion of nitrate to nitrite is catalyzed
by an enzyme called nitrate reductase. This enzyme has several other
important constituents including FAD, cytochrome, NADPH or NADH and
molybdenum.
NO3- + NADH + H+ Nitrate reductase

NO2- + NAD+ + H2O
The overall process of nitrate reduction take place in the cytosol and is an
energy dependent reaction. The enzyme nitrate reductase has been studied in
many plants and it is observed that the enzyme is continuously synthesized and
degraded. The enzyme nitrate reductase is inducible. This means that increase
in nitrate concentration in the cytosol induces more of nitrate reductase to be
synthesized. However, when excess NH4+ is produced then it has a negative
effect on the synthesis of nitrate reductase. In plants, it has also been observed
that light also increases nitrate reductase when nitrate is available.
188 | P a g e
In the second step the nitrite so formed is further reduced to ammonia and this
is catalyzed by the enzyme nitrite reductase. Nitrite present in the cytosol is
transported into chloroplast or plastids where it is reduced to ammonia.
The enzyme nitrite reductase can accept electrons from sources such as
NADH, NADPH or FADH2. Besides, reduced ferredoxin has also been shown
to provide electrons to nitrite reductase for reducing nitrite to ammonia.
Ammonia formed must be utilized quickly by plants because accumulation of
ammonia has a toxic effect. Some plants including algae leach out excess
ammonia which can further be oxidized to nitrite and nitrate by microorganisms
in the soil or water.
Breakdown of Proteins
The first stage in the breakdown of proteins is their hydrolysis to amino acids.
There is a range of proteolytic enzymes which catalyse these reactions.
Some of these are relatively non-specific in their action, but others will only
hydrolyse bonds next to specific, individual amino acids within the protein
sequence. Hydrolysis of proteins takes place in the digestive system and in
cells during protein turnover (balance between protein breakdown and build-
up).
Breakdown of Amino Acids

The amino acids released during digestion are absorbed in the small
intestine and pass to the liver. The liver is the main site of amino acid
metabolism, but kidney also has some capacity to degrade amino acids.
Some amino acids may be metabolized in the liver or may pass to other
tissues where they are used to synthesize new proteins. Any excess amino
acids are oxidized.
Even though each of the 20 amino acids found in proteins is oxidized by a

separate pathway, the first step in their oxidation is usually the removal of
the amino group. This may occur by either transamination or oxidative
deamination.
Transamination
✓ A reversible reaction which is catalysed by transaminases, also known
as aminotransferases, with pyridoxal phosphate (PLP) as cofactor
✓ The alpha amino group of amino acid is transferred to alpha keto acid,
resulting in the formation of a new amino acid and a new keto acid
✓ At the end of the reaction, the donor amino acid becomes new keto acid
and the recipient keto acid becomes new amino acid
189 | P a g e
✓ Although a number of keto acids can take part in this reaction, a-

ketoglutarate is often used and glutamate is produced. This reaction is
catalysed by glutamate transaminase.
✓ Transamination reactions are completely reversible and can also be
used to synthesize amino acids.
✓ Many amino acids can take part in this reaction, their amino groups are
incorporated into glutamate. Thus, the nitrogen from the amino acids is
concentrated as the amino group of glutamate.
Deamination
Because transamination replaces one amino acid by another, it does not
actually result in net amino acid breakdown. Thus, the process of oxidative
deamination is essential for release of ammonia from amino acids. In this
way the alpha amino group of the amino acid is converted directly to
ammonia. The oxidative deamination of glutamate, catalysed by glutamate
dehydrogenase, occurs very commonly and is accompanied by reduction
of NAD+ to NADH and is irreversible. This results in the production of
ammonia (NH3) which is then converted rapidly into urea.
Oxidative deamination of glutamate catalysed by glutamate dehydrogenase
190 | P a g e
Oxidation of Carbon Skeleton of Amino Acids

The α-keto acids produced from the removal of the amino groups from
amino acids by transamination or deamination can be further oxidized by a
series of pathways, resulting in the production of metabolites which include
pyruvate, acetyl-CoA and the TCA-cycle intermediates. They may be
completely oxidized to carbon dioxide, or in some cases, converted into
sugars such as glucose by gluconeogenesis.
In plants, the keto acids formed by breakdown of amino acids are used
mainly in synthesis of new amino acids and are only degraded by oxidation
to a very minor degree.
Summary of amino acid catabolism

Pyruvate or intermediates of the TCA cycle containing three or more carbon

atoms can be converted into oxaloacetate and then into glucose. Hence amino
acids which are broken down into these compounds are called glucogenic
amino acids. On the other hand, compounds such as acetate (in the form of
acetyl CoA or acetoacetyl CoA, which itself breaks down to acetyl CoA) do not
result in oxaloacetate production and therefore cannot be used as precursors
191 | P a g e
of glucose. Instead, the acetyl CoA is converted into a range of other

compounds, including the ketone bodies. Amino acids forming acetyl CoA are
therefore called the ketogenic amino acids. A third group of amino acids break
down into several fragments, some of which are ketogenic and some
glucogenic. These acids must be considered to be both glucogenic and
ketogenic.
The Fate of Ammonia

Ammonia, the product of oxidative deamination reactions, is toxic in even
small amounts and must be removed from the body. Where water is plentiful,
ammonia may be excreted directly, as in protozoa, nematodes and aquatic
amphibians. In most terrestrial animals, ammonia is converted into urea by
the urea cycle, before being excreted. In birds and many terrestrial reptiles,
nitrogen is excreted predominantly as uric acid which is synthesized from
ammonia via the pathway which forms purines
Urea Cycle
The urea cycle begins inside liver mitochondria, but three of the subsequent
steps take place in the cytosol. Urea has two amino groups, one comes
directly from ammonia and the other from the amino group of aspartate.
The ammonia first reacts with carbon dioxide (as HCO3-) and two molecules
of ATP to produce a compound called carbamoyl phosphate. Only one of
the ATPs is needed to provide the phosphate group, the other provides the
energy to drive the reaction. The next four intermediate compounds are all
amino acids but only one of them, arginine, is ever found as part of proteins.
Carbamoyl phosphate reacts with the amino acid, ornithine, to produce

another amino acid, citrulline. This then reacts with a molecule of aspartate
to produce argininosuccinate. This step requires a large amount of energy
so that ATP has to be broken down to AMP and pyrophosphate (PP), using
the equivalent of two high-energy phosphate bonds. The amino group of
the aspartate is derived from glutamate by transamination.
The arginine produced at this stage is then hydrolysed, so that the end of
the molecule is liberated as urea. This regenerates ornithine, with which the
192 | P a g e
cycle began. Hence, both of the amino groups of the urea are ultimately
derived from the amino group of glutamate.
The excretion of ammonia by way of urea requires a considerable amount

of energy. Each turn of the cycle requires the expenditure of four high-
energy phosphate bonds (two ATPs change to ADP and one changes to
AMP). Urea is excreted through the kidneys in the urine. This means that
there is also a high requirement for water in the process.
The urea cycle

193 | P a g e
Biosynthesis of Amino Acids
All amino acids are derived from intermediates in glycolysis, the citric acid
cycle, or the pentose phosphate pathway. Nitrogen enters these pathways
by way of glutamate and glutamine. Some pathways are simple, others are
not. Ten of the amino acids are just one or several steps removed from the
common metabolite from which they are derived. The biosynthetic
pathways for others, such as the aromatic amino acids, are more complex.
Organisms vary greatly in their ability to synthesize the 20 common amino

acids. Whereas most bacteria and plants can synthesize all 20, mammals
can synthesize only about half of them - generally those with simple
pathways. These are the nonessential amino acids, not needed in the diet.
Overview of amino acid

biosynthesis. The carbon skeleton
precursors derive from three
sources:
194 | P a g e
APPLICATION
Answer the following questions:
1. How many molecules of ATP are hydrolysed to form two molecules of

ammonia? ___________________
2. Where do the nitrogen atoms of urea derived from in the urea cycle?
__________________________________
3. What are the products of urea cycle?
___________________________________
4. What is transamination?
___________________________________
5. Differentiate ketogenic from glucogenic amino acids
___________________________________
195 | P a g e
Lesson 4
Lesson Title: Integration of Metabolism
Introduction
Hundreds of reactions simultaneously take place in a living cell. These
reactions are in a well-organized and integrated manner. In the previous
lessons, the different metabolic pathways were discussed. This lesson
covers the interconnectedness of the different pathways.
Learning Outcome
✓ Integrate the overall design of metabolism
ACTIVITY
Examine the figure below and answer the questions in the analysis.
196 | P a g e
ANALYSIS
Answer the following questions:

1. What are the major metabolic pathways?
2. True or False. Studies of metabolism usually separate the reactions into

pathways such as the Krebs' Cycle or the Pentose Phosphate Pathway.
This is really an artificial designation, since these pathways are all
intimately linked and overlap.
3. Which pathway is central to virtually all living organisms?

A) glycolysis.
B) citric acid cycle
C) electron transport chain
D) photosynthesis
E) fatty acid metabolism
4. Which of the following compounds does not overlap between several

pathways?
A) Acetyl-CoA
B) Oxaloacetate
C) Cholesterol
D) Pyruvate
ABSTRACTION
Metabolism is a continuous process, with thousands of reactions,

simultaneously occurring in the living cell. The organisms possess variable
energy demands; hence the supply (input) is also equally variable. The
consumed metabolic fuel may be burnt (oxidized to CO2 and H2O) or stored to
meet the energy requirements as per the body needs. ATP serves as the
energy currency of the cell in this process.
197 | P a g e
Central Themes of Metabolism

Acetyl CoA is a common intermediate for carbohydrate, lipid, and
amino acid metabolism
Oxidation of dietary fuels produce energy captured as ATP and
NADH/FADH2
ATP is the universal energy carrier
NADH and FADH2 transfer electrons to oxygen; the process being
coupled to ATP synthesis
NADPH is the redox agent for reductive biosynthesis
Biomolecules are degraded to smaller constituents; small precursors
are used to build biomolecules
Biosynthetic and degradative pathways are compartmentalized to
make both favorable and energy efficient
Metabolic pathways do not operate separately but usually interact with each
other in coordination manner that serves the total needs of an organism’s
function. Integration of metabolism refers to the coordination between
metabolic pathways inside the body. The anabolic and catabolic pathways are
integrated through a group of common intermediates called common junction
points.
Krebs cycle plays a central role in the integration of metabolism. It contributes

to fatty acid synthesis, amino acid synthesis and gluconeogenesis. Krebs cycle
is the final step in the aerobic catabolism of all nutrient fuels (carbohydrase,
proteins, lipids to CO2 and H2O; thus, the final common metabolic pathway for
the oxidation of all foodstuffs. Also, Krebs cycle is the major source of reducing
198 | P a g e
equivalents, NADH and FADH2, used by the cell to generate ATP via oxidative
phosphorylation.
The main key junction points between metabolic pathways are the
intermediates of:
1. Glucose-6-phospahte – a central in carbohydrate metabolism
2. Pyruvate - acts as important intermediate of carbohydrates and amino
acid metabolism. It links gluconeogenesis and amino acids metabolism
to glycolysis
3. Acetyl CoA - the key and common metabolite, produced from different
fuel sources (carbohydrates, lipids, amino acids). It is central in energy
metabolism
199 | P a g e
Overview of integration of metabolic pathways of energy metabolism

(Source: biology discussion.com)
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.
200 | P a g e
APPLICATION
Explain the metabolic changes that occur during starvation. What appears to
be the principal purpose for the preferential degradation of muscle tissue during
starvation?
Module Summary
Nice one! You have positively completed Module 3. This module has helped
you in expanding your understanding about metabolic processes that
carbohydrates, lipids, and amino acids undergo and the overall design of
metabolism.
Key points covered in the module include:

Metabolism is the set of chemical reactions that occur in a cell, which
enable it to keep living, growing and dividing.
Metabolic processes are classified as catabolism and anabolism
Catabolism refers to the degradation of molecules to provide energy.
Anabolism are reactions using energy to synthesize new molecules for
growth.
The first step of carbohydrate catabolism is glycolysis, which produces
pyruvate, NADH, and ATP.
Under anaerobic conditions, the pyruvate can be converted into lactate
to keep glycolysis working. Under aerobic conditions, pyruvate enters
the Krebs cycle, also called the citric acid cycle or tricarboxylic acid
cycle.
Gluconeogenesis is the synthesis of new glucose molecules from
pyruvate, lactate, glycerol, or the amino acids alanine or glutamine.
Fatty acid oxidation is the process of breaking down fatty acid into acetyl-
CoA units
Amino acids when consumed in excess than required for protein
synthesis are degraded and utilized to meet the fuel demands of the
body
All amino acids are derived from intermediates in glycolysis, the citric
acid cycle, or the pentose phosphate pathway
Acetyl CoA is the key and common metabolite produced from different
fuel sources
The energy trapped in the form of NADH and FADH2 are oxidized in the
ETC and undergo oxidative phosphorylation to generate ATP
201 | P a g e

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NAT SCI 9 BIOCHEMISTRY

Lesson Title: Carbohydrate Metabolism

2) Pre-stretch the balloons. Do this by stretching them and blowing them up

Bottle #1: ½ cup or 118 mL of warm water (40⁰C)

Bottle #2: ½ cup or 118 mL of warm water (40⁰C)

Bottle #3: ½ cup or 118 mL of warm water (40⁰C)

Bottle #:4 ½ cup or 118 mL of warm water (40⁰C)

6) Do the same for the replicates.

7) Fill out the data table.

Guide Questions for Discussion:

1) What process was observed in this lab?

3) What is the overall equation of cellular respiration?

Metabolism is the total of all chemical reactions in an organism. The reaction

In anabolic reactions, simple organic molecules such as pyruvic acid, acetyl

In catabolic reactions, complex substances are broken down to simpler

I of catabolism is the breakdown of food molecules by hydrolysis

3. Stage III - oxidation of the reducing equivalents by oxygen with the

Digestion, absorption, and transport of Carbohydrates

respectively. The major products of the complete hydrolysis of disaccharides

The monosaccharides (glucose, galactose, and fructose) are taken up

Transport of glucose across the intestinal epithelium

Major Pathways in Carbohydrate Metabolism

Importance of the glycolytic pathway:

The glycolytic pathway

2. Stage 2 (Energy yielding phase)

The glycolytic pathway

Step 1. Phosphorylation of Glucose

Source: Lehninger Principles of Biochemistry

Step 2. Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate

Source: Lehninger Principles of Biochemistry

Step 3. Phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate

Source: Lehninger Principles of Biochemistry

Step 4. Cleavage of fructose 1,6-bisphosphate

Source: Lehninger Principles of Biochemistry

Step 5. Interconversion of the Triose Phosphates

Source: Lehninger Principles of Biochemistry

Step 6. Oxidation of Glyceraldehyde 3-Phosphate to 1,3-Bisphosphoglycerate

Source: Lehninger Principles of Biochemistry

Step 7. Phosphoryl Transfer from 1,3-Bisphosphoglycerate to ADP

Source: Lehninger Principles of Biochemistry

Step 8. Conversion of 3-Phosphoglycerate to 2-Phosphoglycerate

Source: Lehninger Principles of Biochemistry

Step 9. Dehydration of 2-Phosphoglycerate to Phosphoenolpyruvate

Source: Lehninger Principles of Biochemistry

Step 10. Transfer of the Phosphoryl Group from Phosphoenolpyruvate to ADP

Source: Lehninger Principles of Biochemistry

The entry of other sugars in glycolytic pathway

It is not difficult to see how fructose enters glycolysis, it merely needs to be

fructose + ATP fructose-6-phosphate + ADP

In animals, hexokinase can phosphorylate fructose as well as glucose.

The entry of galactose is not as simple as that of fructose: it must be converted

Galactose is phosphorylated by galactokinase to give galactose 1-phosphate.

Galactose 1-phosphate uridylyl transferase catalyzes the transfer of

Finally, the glucose 1-phosphate is converted to glucose 6-phosphate by

Conversion of galactose to glucose 1-phosphate

D-Mannose, released in the digestion of various polysaccharides and

Mannose 6-phosphate is isomerized by phosphomannose isomerase to yield