Carbohydrates

Carbohydrates are one of the essential food ingredients , which we all require. Every one of
us is quite familiar with the word glucose and cane sugar . They are simple carbohydrates and
we consume carbohydrates in one form or every other meal we take. They are primary source
of energy in the body and constitute an important part of any well balanced diet.
Bread, rice, potatoes, peas etc all contains carbohydrates the sweetening agents includes
nothing but simple carbohydrates.

Biological Importance:
Carbohydrates are the most widespread organic compounds occurring in nature. they are
present in almost all the plants and comprise about 80% of dry weight. The most important
are cellulose-the chief structural material of plants, starches and sugars-sucrose and glucose.
in animals and human beings glucose is an essential constituent of blood and takes part in
various metabolic reactions carbohydrates occur in bound form in biologically important
compounds like which is a key material in biological energy storage and transport system.
ATP is called energy currency of the cell. carbohydrates are also present in nucleic acids
which control the production of enzymes and transfer genetic information. Carbohydrates are
major source of energy . Our body can mobilize carbohydrates more easily than fat , even
though fats contain more energy on a gram for gram basis. Free glucose in the bloodstream is
quickly depleted but we have stored glucose in readily accessible for as polymer glycogen.
when we require energy glycogen breakdown into glucose in a quick response to need for
energy. oligosaccharide play a key role in processes that take part on the surface of cells.
polysaccharide such as cellulose are essential components of grass ,trees and other
polysaccharide are major constituents of bacterial cell walls. starch is the principal food
reserve of plant.

1. Carbohydrates provides energy and regulation of blood glucose.

2. It will prevent the degradation of skeletal muscle and other tissues such as the heart, liver, and
kidneys.
3. It prevents the breakdown of proteins for energy.
4. Carbohydrates also help with fat metabolism. If the body has enough energy for its immediate
needs, it stores extra energy as fat.
5. Carbohydrates are an important component of many industries like textile, paper, lacquers and
breweries.
6. Detoxification of physiological importance is carried out to some extent with carbohydrate
derivatives.
7. Agar is polysaccharide used in culture media, laxative and food.
8. Carbohydrates form a part of genetic material like DNA and RNA in the form of deoxyribose and
ribose sugars.
9. Hyaluronic acid found in between joints acts as synovial fluid and provides frictionless movement.
10. They help make up the body mass by being included in all the parts of the cell and tissues.
11. Adequate storage of hepatic glycogen helps in detoxifying a normal liver.
12. They form components of bio-molecules which have a key role in blood clotting, immunity,
fertilization etc.
13. Carbohydrates is basically the main fibre of the diet or provide the bulk fibre for better digestion.
14. Carbohydrates help clear gut and prevent constipation.
15. Starch is the form the food is stored in plants.

16. It provides sweetness to foods.

17. Pectin and Hemicellulose are the structural carbohydrate in plant cell walls.
18. It plays important roles in cellular recognition processes.
19. Chitin forms the cell wall of fungi and the outer schelitone of insects.
20. Murine is a structural carbohydrate in bacterial cell wall.

Originally the name carbohydrate was given to all such compounds having the general formula
Cx(H2O)y i.e. , They were considered to be hydrates of carbon. now we know that carbohydrates
are not simply hydrates of carbon but have a variety of other structural features. They are usually
defined as Poly hydroxy aldehyde and ketones or substances that hydrolyse to yield Poly hydroxy
aldehyde and ketones. Poly hydroxy aldehydes are also called aldoses and Poly hydroxy ketones are
also called ketoses. Low molecular weight carbohydrates are also called as sugar or saccharides
example glucose sucrose are simple sugars. Common sugars like glucose fructose and some less
common sugars also occurs in the combined state with various hydroxy compounds. such derivatives
are called glycosides and non-sugar component is called aglycone. When we have a sugar component
(glucose) the compound is called glucoside, if fructose is present it is a fructoside and so on.
Glycosides are widely distributed in plants and animals. Structurally, these compounds are related to
simple methyl glucosides. When the sugar moiety forms an ether linkage with the aglycone. It is
called O-glycoside and when there is a formation of a C-C bond, it is called C-glycoside.

Carbohydrates are produced in green plants as a result of photosynthesis. it involves chemical

combination or fixation of carbon dioxide and water by utilization of light energy. in this process
water is oxidised to oxygen and carbon dioxide is reduced. the overall equation for photosynthesis can
be written as.

Besides providing energy, a major metabolic product of glucose is acetyl Coenzyme A,

which acts as a starting point for synthesis of fats, fatty acids, amino acids and large number
of other important compounds required by our body.

CLASSIFICATION OF CARBOHYDRATES
Carbohydrates are classified on the basis of number of aldose and/ or ketoses units produced upon
hydrolysis.
Metabolism of Carbohydrates

Carbohydrates are organic molecules composed of carbon, hydrogen, and oxygen atoms. The family
of carbohydrates includes both simple and complex sugars. Glucose and fructose are examples of
simple sugars, and starch, glycogen, and cellulose are all examples of complex sugars. The complex
sugars are also called polysaccharides and are made of multiple monosaccharide molecules.
Polysaccharides serve as energy storage (e.g., starch and glycogen) and as structural components
(e.g., chitin in insects and cellulose in plants).

During digestion, carbohydrates are broken down into simple, soluble sugars that can be transported
across the intestinal wall into the circulatory system to be transported throughout the body.
Carbohydrate digestion begins in the mouth with the action of salivary amylase on starches and
ends with monosaccharides being absorbed across the epithelium of the small intestine. Once the
absorbed monosaccharides are transported to the tissues, the process of cellular respiration begins

Metabolic pathways
Metabolic pathways and cycles are reaction chains where chemical products become the substrate for
the next step. All substrates are chemically transformed in reactions that belong to either pathways (if
the reactions are aligned in linear fashion) or metabolic cycles (if the moieties of the reactions are
preserved). The term substrate oxidation is used for substrate degradation ultimately leading to
CO2 production.

 A metabolic pathway is a series of chemical reactions in a cell that build and breakdown
molecules for cellular processes.
 Anabolic pathways synthesize molecules and require energy.
 Catabolic pathways break down molecules and produce energy.
 Because almost all metabolic reactions take place non-spontaneously, proteins called enzymes
help facilitate those chemical reactions.

The processes of making and breaking down carbohydrate molecules illustrate two types of metabolic
pathways. A metabolic pathway is a step-by-step series of interconnected biochemical reactions that
convert a substrate molecule or molecules through a series of metabolic intermediates, eventually
yielding a final product or products. For example, one metabolic pathway for carbohydrates breaks
large molecules down into glucose. Another metabolic pathway might build glucose into large
carbohydrate molecules for storage. The first of these processes requires energy and is referred to as
anabolic. The second process produces energy and is referred to as catabolic. Consequently,
metabolism is composed of these two opposite pathways:

1. Anabolism (building molecules)

2. Catabolism (breaking down molecules)

Anabolic Pathways

Anabolic pathways require an input of energy to synthesize complex molecules from simpler ones.
One example of an anabolic pathway is the synthesis of sugar from CO2. Other examples include the
 synthesis of large proteins from amino acid building blocks and the synthesis of new DNA strands
from nucleic acid building blocks. These processes are critical to the life of the cell, take place
constantly, and demand energy provided by ATP and other high-energy molecules like NADH
(nicotinamide adenine dinucleotide) and NADPH.

Catabolic Pathways

Catabolic pathways involve the degradation of complex molecules into simpler ones, releasing the
chemical energy stored in the bonds of those molecules. Some catabolic pathways can capture that
energy to produce ATP, the molecule used to power all cellular processes. Other energy-storing
molecules, such as lipids, are also broken down through similar catabolic reactions to release energy
and make ATP.

Anabolism
Catabolism

Catabolism breaks down big complex molecules Anabolism builds molecules required for the
into smaller, easier to absorb molecules. body’s functionality.

The process of catabolism releases energy. Anabolic processes require energy.

Hormones involved in the process are

Hormones involved in the processes are
estrogen, testosterone, growth hormones
adrenaline, cytokine, glucagon, and cortisol.
and insulin.

Examples of catabolic processes are proteins Examples include the formation of

becoming amino acids, glycogen breaking down polypeptides from amino acids, glucose
into glucose and triglycerides breaking up into forming glycogen and fatty acids forming
fatty acids. triglycerides.

In catabolism, potential energy is changed into In anabolism, kinetic energy is converted

kinetic energy. into potential energy.

It is required to perform different activities in living It is required for maintenance, growth, and
entities. storage.

Requires NADPH Requires NAD+

Endergonic Exergonic

The free energy released by catabolic processes is conserved through synthesis of ATP from ADP and
phosphate or through reduction of coenzyme NADP + to NADPH.
ATP and NADPH are the major free energy sources for anabolic pathways.
Degradative metabolism is that converts diverse complex substances (carbohydrates, lipids and
proteins) to common intermediates. These intermediates are then further metabolized in a central
oxidative pathway that terminates in a few end products.
Biosynthesis carries out the opposite process, relatively few metabolites mainly pyruvate, acetyl
coenzymeA and citric acid cycle intermediates serve as starting material for a host of varied
biosynthetic products.
Metabolic pathways have following principal characteristics:
1. Metabolic pathways are irreversible.
2. If two metabolites are interconvertible, the synthetic route from first to second
must differs from second to first.
3. Every metabolic pathway has exergonic first committed step.
4. All metabolic pathways are regulated, usually at the first committed step.
5. Metabolic pathways in eukaryotes occurs in specific subcellular
compartments.

CoFactor
A cofactor is a non-protein chemical compound or metallic ion that is required for an enzyme's
activity as a catalyst (a catalyst is a substance that increases the rate of a chemical
reaction). Cofactors can be considered "helper molecules" that assist in biochemical transformations.

Cofactor, a component, other than the protein portion, of many enzymes. If the cofactor is removed
from a complete enzyme (holoenzyme), the protein component (apoenzyme) no longer has catalytic
activity. A cofactor that is firmly bound to the apoenzyme and cannot be removed without
denaturing the latter is termed a prosthetic group; most such groups contain an atom of metal such
as copper or iron. A cofactor that is bound loosely to the apoenzyme and can be readily separated
from it is called a coenzyme. Coenzymes take part in the catalysed reaction, are modified during the
reaction, and may require another enzyme-catalysed reaction for restoration to their original state.

A coenzyme is an organic non-protein compound that binds with an enzyme to catalyze a

reaction. Coenzymes are often broadly called cofactors, but they are chemically different. A
coenzyme cannot function alone, but can be reused several times when paired with an enzyme .

NAD + , or nicotinamide adenine dinucleotide, is a critical coenzyme found in every cell in your
body, and it’s involved in hundreds of metabolic processes. But NAD + levels decline with age.
NAD + has two general sets of reactions in the human body: helping turn nutrients into energy as
a key player in metabolism and working as a helper molecule for proteins that regulate other
cellular functions.
NADH is the abbreviation for the naturally occurring biological substance, nicotinamide adenine
dinucleotide hydride. The “H” stands for high-energy hydrogen and indicates that this substance is in the
most biologically active form possible. Often referred to as coenzyme 1
NADH is also an important antioxidant; in fact, scientists acknowledge that NADH is the most powerful
antioxidant to protect cells from damage by harmful substances
GLYCOLYSIS
In plants energy stored in the form of starches while glycogen is the storage polysaccharide present in
animals . Glycogen is found in animal cells in granule similar to the starch granules in plant cells .
Glycogen granules are observed in well fed liver and muscle cells.
glycogen is the branch chain polymer of Alpha D glucose and in the respect it is similar to
amylopectin fraction of starch . packed in glycogen consist of Alpha (1→6) linkages with Alpha
(1→6) linkage at branch points however glycogen is more highly blanched than amylopectin . Branch
point occurs about every 10 residue in glycogen and every 25 residue in amylopectin. in glycogen
every chain length is 13 glucose residue there are 12 layers of branching.
When we digest a meal high in carbohydrates , we have a supply of glucose that exceeds our
Immediate needs. therefore excess of glucose is stored in our body in the form of glycogen primarily
in the liver and muscles. when we need energy, various degradative enzymes breakdown glycogen to
smaller polysaccharides or free glucose units. in muscle glucose 6 phosphate obtained from glycogen
breakdown, enters the glycolytic pathway directly rather than being hydrolysed glucose first. however
liver glycogen breaks down to glucose-6-phosphate which is hydrolysed to give glucose. the release
of glucose from liver by this breakdown of glycogen replenishes the supply of glucose in the blood.
The conversion of glycogen to glucose-6 phosphate takes place in 3 steps:

1.In the first reaction each glucose residue clean from glycogen reacts with the inorganic phosphate
to give glucose 6 phosphate. this cleavage reaction is phosphorolysis and not hydrolysis. the enzyme
that catalyse this reaction is known as glycogen phosphorylase, which cleaves one glucose unit at a
time from non-reducing end of a branch glycogen to produce glucose one phosphate and reminder
of the glycogen molecule. this then enter the metabolic pathways of carbohydrate breakdown.

2.In the second reaction glucose one phosphate isomerase to give glucose-6phosphate by
the enzyme phosphoglucomutase.

Complete breakdown of glycogen also requires de- branching enzymes that degrade alpha (1to6)
Linkages and thereby expose additional 1to6 glycosidic linkages to be attacked by phosphorylase.
Glycolysis is the first stage of glucose metabolism. it is an anaerobic process and gives only small
amount of energy in the form of only 2 molecules of ATP. we will see the complete aerobic oxidation
of glucose to carbon dioxide and water via citric acid cycle yields high energy equivalent of 30-32
molecules of ATP.

After the formation of glucose-6-phosphate, glycolysis and alcoholic fermentation follow a common
pathway under anaerobic conditions until pyruvate is formed. In, glycolysis, pyruvate is converted to
lactic acid, which will eventually be exported from the muscle to liver.
When pyruvate is formed, it can have one of the several fates:
Under aerobic conditions, pyruvate loses carbon dioxide and the remaining 2 carbon Atom become
link to coenzyme A acetyl group to form acetyl-CoA which then enters the citric acid cycle. There are
two fates of pyruvate in anaerobic metabolism.
in organisms capable of alcoholic fermentation pyruvate loses carbon dioxide producing
acetaldehyde which in turn reduce to give ethanol.
in the second anaerobic metabolism lactate is produced especially in the muscles. This is anaerobic
glycolysis. the conversion of glucose to pyruvate is simply called glycolysis.
Given below is the conversion of pyruvate to dictate in the muscle:
This is the final reaction of anaerobic glycolysis pyruvate is reduced to lactic in the presence of
NADH. electric dehydrogenase is the enzyme that catalyses this reaction

the NADH produced from NAD+ by earlier oxidation of glyceraldehyde 3 phosphate is used up with
no net change in the relative amounts of NADH and NAD+ in the cell this regeneration is required
under anaerobic conditions in the cell so that NAD+ will you present for further glycolysis to take
place.

Control points in glycolysis

Pathways can be shut down if an organism has no immediate need for their products. This saves
energy for the organism.
In glycolysis, there are three control points.
1.The first is reaction of glucose to glucose-6phosphate which is catalysed by hexokinase. This
conversion is inhibited by glucose-6phosphate itself.
2.The second control point is the production of fructose-1,6-bisphosphate.This step is catalysed by
phosphofructokinase and is inhibited by ATP
3.The last point is the conversion of phosphoenol pyruvate to pyruvate, catalysed by pyruvate
kinase. This step is also inhibited by ATP
Alcoholic Fermentation
Ethanol fermentation, also called alcoholic fermentation, is a biological process which converts
sugars such as glucose, fructose, and sucrose into cellular energy, producing ethanol and carbon
dioxide as by-products.
(Because yeast performs this conversion in absence of oxygen ethanolic fermentation is classified
as anaerobic.)

Overall Pathway in Alcoholic Fermentation

In alcoholic fermentation one molecule of glucose is converted to fructose-1,6-diphosphate. which
eventually give rise to 2 molecules of pyruvate. pyruvate loses carbon dioxide producing
acetaldehyde which in turn is reduced to give ethanol.
All these reaction can be divided into three phase.
Step 1. Glucose is phosphorylated to give glucose-6-phosphate
The enzyme that catalyse this reaction is hexokinase. this reaction uses ATP and gets converted into
ADP. the term kinase is applied to the class of ATP dependent enzymes that transfer a phosphate
group from ATP to a substrate. the substrate of hexokinase is not necessarily glucose; rather it can
be any number of hexose such as glucose fructose and mannose. Mg2+ is cofactor.

(A cofactor is a non-protein chemical compound or metallic ion that is required for an enzyme's
activity as a catalyst (a catalyst is a substance that increases the rate of a chemical
reaction). Cofactors can be considered "helper molecules" that assist in biochemical
transformations).

Step 2. Glucose-6-phosphate isomerises to give fructose-6-phospahte

Phosphoglucose isomerase is the enzyme that catalyse this reaction. The C-1 aldehyde group of
glucose-6-phosphate is reduced to hydroxyl group and C-2 hydroxyl group is oxidized to give ketone
of fructose-6-phosphate.

Step3. fructose 6 phosphate is further phosphorylated producing fructose-1,6-biphosphate.

this reaction takes place in the presence of ATP. phosphofructokinase is the enzyme and Mg 2+ as a
cofactor. after fructose-1,6-biphosphate is formed from original sugar, no other pathways are
available and the molecule must undergo rest of the reactions of alcoholic fermentation this
reaction is highly exothermic and irreversible.

Step 4. Fructose-1,6-bisphospahte is split into two 3-carbon fragments.

The cleavage reaction here is reverse of an aldol condensation. The enzyme that catalyses it is called
aldolase. Fragments(Dihydroxy-acetone-phosphate, D-Glyceraldehyde-3-phospahte).

Step 5. The dihydroxyacetone phosphate is converted to glyceraldehyde-3-phosphate.

the enzyme that catalyses the reaction is triosephosphate isomerase. one molecule of
glyceraldehyde has already been produced by aldolase reaction. we now have a second molecule of
glyceraldehyde 3 phosphate produced in this reaction. the original molecule of glucose which
contains 6 carbon atoms, has now been converted to 2 molecules of glyceraldehyde- 3-phosphate
which contains 3 carbon atoms.

In the first 5 steps one molecule of glucose is split into two 3 carbon compounds that is
D-glyceraldehyde-3-phosphate. Two molecules of ATP are required for these reactions.

#Up till now glucose has been converted to two molecules of glyceraldehyde 3 phosphate. we have
not seen any oxidation reaction yet. in this phase of pathway, ATP is produced instead of being used.

Step 6. The oxidation of glyceraldehyde 3 phosphate to 1,3-biphosphoglycerate.

Here NAD+ is the coenzyme. The reaction involves the addition of a phosphate group to
glyceraldehyde-3-phosphate as well as electron-transfer reaction from glyceraldehyde-3-phosphate
as well as electron transfer reaction from glyceraldehyde-3-phosphate to NAD+ i.e; NAD+ is reduced
to NAD.

NAD+ +2H+ +2e-→ NADH + H+

The enzyme that catalyses this conversion is glyceraldehyde-3-phosphate dehydrogenase
Step 7. In this step ATP is produced by phosphorylation of ADP and 3
phosphoglycerate is formed.
The enzyme that catalyses reactions called phosphoglycerate kinase

Step 8.The phosphate group is transferred from carbon 3 to carbon 2 of the glyceric
acid backbone
The enzyme that cut ties this reaction is phosphoglyceromutase.
Step 9. The 2-phosphoglycerate loses one molecule of water producing phosphoenol
pyruvate
This reaction does not involve electron transfer , it is a dehydration reaction. enolase is the
enzyme that catalyses the reaction, requires Mg2+ as a cofactor. the water molecule that is
eliminated binds to Mg2+in the course of reaction.

Step 10. Phosphoenolpyruvate transfers it's phosphate group to ADP, producing

pyruvate
Here double bond shifts to oxygen on carbon 2 and hydrogen shifts to carbon 3.
pyruvate kinase is the enzyme that catalyse this reaction.

Thus in the second phase of alcoholic fermentation glyceraldehyde 3 phosphate is converted

to pyruvate. these reactions yields 4 molecules of ATP two for each molecules of pyruvate
produced.
The conversion of pyruvate to acetaldehyde and then to ethanol
Pyruvate is decarboxylated to produce acetaldehyde. the enzyme that catalyse this reaction is
pyruvate decarboxylase. this enzyme requires Mg2+ and co-factor thiamine pyrophosphate
(TPP). Carbon dioxide splits off leaving a two carbon fragment covalently bonded to TPP.
Thus acetaldehyde is produced. This step is irreversible.

The carbon dioxide produced is responsible for the bubbles in the beer and in sparkling
wines.
Acetaldehyde is then oxidized to produce within all and at the same time one molecule of
NADH is oxidized to NAD+, for each molecule of ethanol produced.

This step provides recycling of NAD+ and thus allows further anaerobic reactions.
The net reaction for alcoholic fermentation is

NAD+ and NADH do not appear in net equation. It is essential that recycling of NADH to
NAD+ takes place at this step so that further anaerobic oxidation takes place. Alcohol
dehydrogenase is the enzyme that catalyses the conversion of acetaldehyde to ethanol. It is
NADH-linked dehydrogenase.
Coenzyme A (CoA, CoASH, or HSCoA) is a coenzyme, well known for it's role in the
synthesis and oxidation of fatty acids, and the oxidation of pyruvate in the citric
acid cycle. All genomes sequenced to date encode enzymes that use coenzyme A as a
substrate, and around 4% of cellular enzymes use it, or a thioester form of it, as a
substrate.

CITRIC ACID CYCLE

Glycolysis release only a very small amount of chemical energy which is available in the
glucose molecule. Much more energy is released when glucose molecule is oxidised
completely to CO2 and water. This is aerobic metabolism. Organisms can obtain far more
energy from nutrients by aerobic oxidation than anaerobic oxidation. We know that
glycolysis produces only two molecules of ATP for each molecule of glucose metabolised.
However, in complete aerobic oxidation each molecule of glucose produces about 30 to 32
molecules of ATP.
Three processes play roles in aerobic metabolism: the citric acid cycle, electron transport and
oxidative phosphorylation.
Metabolism consists of catabolism which is the oxidative breakdown of nutrients and
anabolism which is reductive synthesis of biomolecules. The citric acid cycle is amphibolic
i.e., it plays a role in catabolism and anabolism. The citric acid cycle, as the hub of metabolic
pathways, serves to connect the breakdown and synthesis of proteins, carbohydrates and
lipids. Most of the metabolites of major nutrients can be fed into the citric acid cycle as
acetyl-CoA and then oxidised to produce energy.
The citric acid cycle operates inside the mitochondrion which acts as its engine room. Here,
metabolic fuels-glucose derived from carbohydrates, amino acids derived from protein and
fatty acids from lipids-are fed into the cycle and will ultimately be oxidised to carbon dioxide
and water. The energy is transferred to electron carriers and finally to the terminal electron
acceptor-oxygen.
There are two other common names for the citric acid cycle. One is Krebs cycle and other
name is tricarboxylic acid cycle(or TCA cycle) because of acids with three carboxyl groups.

OVERALL PATHWAY OF THE CITRIC ACID CYCLE

The citric acid cycle takes place in mitochondria. Most of the enzymes of the citric acid cycle
are present in the mitochondrial matrix.
Pyruvate produced by glycolysis is oxidized to one carbon dioxide molecule and one acetyl
group which becomes linked to coenzyme A to give acetyl-CoA. It is then enters the citric
acid cycle. Two more molecules of carbon dioxide produced for each molecule of acetyl-CoA
that enters the cycle and electrons are transferred in the process. electronic accepter in all
cases but one is NAD+, which is reduced to NADH. in one case electron acceptor is FAD
(flavin adenine dinucleotide ). this takes up two electrons and two hydrogen ions to produce
FADH2. The electrons are passed from NADH and FADH2 to several stages and final
acceptor is oxygen with water as the product.

There are 8 step in the citric acid cycle, each catalysed by a different enzyme. Four of the 8
steps -steps 3, 6 , 6 and 8- are oxidation steps.

The oxidizing agent is NAD+ in all except step 6, in which it is FAD. In step 5 , a molecule
of GDP (guanosine diphosphate) is phosphorylated to produce GTP.

in the first reaction of the cycle, the two carbon acetyl group condenses with four carbon
oxaloacetate to produce six carbon citrate ion. in the next few steps citrate isomerises and
then oxidative decarboxylation takes place to give Alpha-keto-glutarate, which again is
oxidatively decarboxylated to produce four-carbon compound succinate. the cycle is
completed by regeneration of oxaloacetate from succinate via the fumarate in several steps.

Conversion of pyruvate to acetyl-CoA

Pyruvate is generated from glycolysis under anaerobic conditions. It moves from the cytosol
in the mitochondrion via specific transporter. Here pyruvate is converted to carbon dioxide
and the acetyl portion of acetyl-CoA. An oxidation reaction precedes the transfer of acetyl
group to the CoA.

The whole process involves several enzymes, all of which are part of pyruvate dehydrogenase
complex. the overall reaction is

Pyruvate + CoA-SH + NAD+ → Acetyl-CoA + CO2+ H+ +NADH

There is -SH group at one end of CoA, it is frequently shown in equations as CoA-SH. The
reaction is catalysed by four cofactors, TPP, FAD, Mg2+ and lipoic acid.

Step 1. Formation of Citrate:

This is the first step of citric acid cycle. it involves reaction of acetyl-CoA and oxaloacetate to form
citrate and CoA-SH. it is a condensation reaction because a new carbon-carbon bond is formed.
reactions catalysed by enzyme citrate synthase.
Step 2. Isomerisation of Citrate to Isocitrate:
This reaction is catalysed by aconitase. the enzyme requires Fe2+ as cofactor. the enzyme is
able to select one end of the citrate molecule in preference to the other.

Step 3. Formation of 𝜶- ketoglutarate and CO2 (First Oxidation Step)

This is the oxidative decarboxylation of isocitrate to Alpha ketoglutarate and carbon dioxide.
the enzyme catalysis this reaction is isocitrate dehydrogenase. this is the first reaction in
which NADH is produced.
Step 4. Formation of Succinyl-CoA and CO2 (Second oxidation step)
This reaction occurs in several stages and is catalysed by enzyme called 𝛼-ketoglutarate
dehydrogenase complex. The co-factors used are TPP, FAD, Mg2+ and lipoic acid.

Step 5. Formation of Succinate

In this step succinyl-CoA is hydrolysed to produce succinate and CoA-SH. the reaction is
catalysed by enzyme succinyl-CoA synthetase. the accompanying reaction is phosphorylation
of GDP to GTP. the energy required for phosphorylation of GDP to GTP is provided by the
hydrolysis of succinyl-CoA to produce succinate and CoA.

Step 6. Formation of Fumarate (Oxidation Step)

Succinate is oxidized to fumarate, reaction is catalyzed by enzyme succinate dehydrogenase. in
this reaction, FAD is the Co enzyme which is reduce to FADH2.
Step 7. Formation of L-Malate
This reactions catalyzed by enzymes fumarase. water is added across the double bond of
fumarate in a hydration reaction to give malate. here the reaction is stereospecific. it gives only
L-Malate.

Step 8. Regeneration of Oxaloacetate (Final Oxidation Step)

This is the final step in which malate is oxidized to oxaloacetate. This reaction catalysed by
maleate dehydrogenase. Another molecule of NAD+ is reduced to NADH. the oxaloacetate
produced can then react with another molecule of Acetyl-CoA to start another round of cycle.

The oxidation of pyruvate by the pyruvate dehydrogenase complex and the citric acid results in
the production of 3 molecules of CO2. during this. One molecule of GDP is phosphorylated to
GTP, one molecule of FAD is reduced to FADH2 and four molecules of NAD+ are reduced to
NADH. Three molecules of life NADH come from the citric acid cycle and one comes from the
reaction of the pyruvate dehydrogenase complex.

From pyruvate dehydrogenase complex

Pyruvate + CoA-SH + NAD+ → Acetyl-CoA + CO2+ H+ +NADH
From citric acid cycle