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 Diploma in Pharmacy 2nd Year
Biochemistry & Clinical Pathology
Chapter 8 : Metabolism
Topics Page No
Metabolism (Study of cycle/pathways without
chemical structures)
Metabolism of Carbohydrates : Glycolysis, 3
TCA cycle and glycogen metabolism, regulation
of blood glucose level. Diseases related to
abnormal metabolism of Carbohydrates
Metabolism of lipids : Lipolysis, β-oxidation of 4
Fatty acid (Palmitic acid) ketogenesis and
ketolysis. Diseases related to abnormal
metabolism of lipids such as Ketoacidosis, Fatty
liver, Hypercholesterolemia
Metabolism of Amino acids (Proteins) : 6
General reactions of amino acids and its
significance– Transamination, deamination,
Urea cycle and decarboxylation. Diseases related
to abnormal metabolism of amino acids,
Disorders of ammonia metabolism,
phenylketonuria, alkaptonuria and Jaundice.
Biological oxidation : Electron transport chain
and Oxidative phosphorylation
 Biochemistry & clinical pathology
Chapter 8
Metabolism
 Different types of chemical reactions that occurs in a living organism is called metabolism.

Major types of Metabolism

 Catabolism : Break Down of molecules to obtain energy.
 Anabolism : Synthesis of any compound needed by body.

Metabolism of Carbohydrates
 Biochemical processes involved in synthesis, breakdown, and inter-conversion of
carbohydrates in living organisms, are collectively called as carbohydrate metabolism
 Glucose (a monosaccharide metabolised by nearly all organisms) is the essential molecule of
carbohydrate metabolism which participates in various metabolic pathways. Insulin is the
primary metabolic hormone synthesised in pancreas and regulates glucose level in blood.
 The metabolism of carbohydrate is simplest than other like protein , and fat , that is why it is
used as immediate source of energy.
 The unused glucose stored in the liver in the form of Glycogen.

Some Pathways of Metabolism of Carbohydrate

 Glycolysis : Break Down of glucose into ATP ( energy ) and Pyruvate and Lactate.
 Citric Acid Pathway or Cycle ( Krebs Cycle , Tricarboxylic Acid Cycle ( TCA) ) : It is the
common metabolic Pathway for Carbohydrates , Fats and protein Oxidation.
 Glycogenesis : In this pathway , The glycogen is synthesised from Glucose.
 Glycogenolysis : In this pathway glycogen is converted into Glucose.
 Gluconeogenesis : In this pathway glucose is Synthesised from a non- carbohydrate source , (
fats and proteins )

Glycolysis
 Glycolysis is an important pathway of Carbohydrate metabolism and it occurs in the Cytoplasm
of a living cell.
 Glycolysis is important to maintain ATP balance.
 The Enzymes involved in Glycolysis are present in Cytosol of cell.
Citric Acid Pathway or Cycle ( Krebs Cycle , Tricarboxylic Acid
Cycle ( TCA) ) :
 It is the common metabolic Pathway for Carbohydrates , Fats and protein .
 First time , Hans Crebs had given the reaction of Citric Acid Cycle , that is why it is called
Crebs Cycle.
 The end product of glycolysis Pyruvate , enters into mitochondria by Active Transport , and
Converted into Acetyl Co-A .
 Now Acetyl Co-A enters into Crebs Cycle , which occurs in the matrix of Mitochondria .
 When one crebs cycle completed in the presence of optimum amount of O2 12 ATPs are
generated .
 Catabolism of one glucose in aerobic Condition ( if O2 is present in mitochondria ) gives 38
ATP , including :
 8 ATPs in Glycolysis
 6 ATPs in conversion of Pyruvate to Acetyl-CoA .
 24 ATPs in Crebs cycle .
 Glycogen Metabolism
 Glycogen is a highly branched polysacharide ( polymer of carbohydrates ) , and 8-10 glucose
units present at per branch.
 Glucose is stored in animal in the form of Glycogen.
 Glycogen is mainly stored in liver (6-8 %) and in muscles cells (1-25 )
 According to NCBI about (500g) glycogen stored in muscles and (100g) in liver.
 Glycogen stored in the form of granules in cell cytosol , where most of the enzymes are found
which required for glycogen synthesis and breakdown.
 First of all liver glycogens are consumed for energy.

Glycogen Metabolism is also in two types :

 Glycogenesis
 Glycogenolysis

Glycogenesis
 The formation ( synthesis ) of glycogen from glucose is called Glycogenesis . Glycogen is a
highly branched polysacharide ( polymer of carbohydrates ) , and 8-10 glucose units present at
per branch .
 Glycogenolysis
 The process of conversion of stored Glycogen into glucose is called glycogenolysis.

Regulation of Blood glucose Level

 The normal blood Glucose level is necessary for proper functioning of body's organs.
 The blood glucose level is regulated by two hormones Insulin and Glucagon , both are secreted
by Pancreas.

Insulin
 The ß cells of pancreas secret insulin hormone.
 The secretion of insulin increases when concentration of glucose increased in blood , and its
secretion deceases when concentration of glucose decreased.
 Insulin helps the glucose for entering into cells , by this way increase the consumption of
glucose.
 A fasting glucose level of 99 mg/dl or less is normal . but less than 70 mg/dl is not normal (
may lead to hypoglycemia )
Glucagon
 The α cells of pancreas secrets glucagon.
 The secretion of glucagon increases when concentration of glucose decreases in blood , and its
secretion decline when blood glucose level increased.
 The glucose stored in liver in the form of glycogen , released in blood under influence of
glucagon.
 Glucagon also stimulates the liver and other cells to synthesis glucose from proteins and fats,
when it required.

Diseases Related to Abnormal Metabolism of Carbohydrates

1) Diabetes Mellitus
 Diabetes Mellitus is a inherited or acquired disease occurs due to defect in insulin secretion or
insulin action or both , in which blood sugar level is high for long time.
 It is also called Hyperglycemia.

Symptoms
 Presence of sugar in urine
 Increase thirst
 Increase frequency of urination
 extreme hunger
 fatigue
 blurred vision
 Headache
 frequent infection
 delay in healing of cuts and wounds
 Itchy skin

2) Galactosaemia
 It is genetic disorder in which body unable to metabolise galactose sugar , and blood galactose
level increased. The absence or non-Functionality of Galactose - 1- Phosphate
Uridylyltransferase Enzyme is responsible for this disorder.

Symptoms
 Appetite loss
 Jaundice
 Enlargement of Liver
 Liver damage
 Abdominal swelling due to fluid accumulation
3) Glycogen Storage Disease (GSD )
 The breakdown of glycogen to glucose is facilitated by some enzymes , if they gets blocked ,
the glycogen accumulate in the liver and muscles and Glycogen Storage Disease developed.

4) Glucose -6- Phosphate Dehydrogenase Deficiency Disease

 It is a genetic disorder in which the quantity of Glucose -6- Phosphate Dehydrogenase
decreased and causes breakdown of Premature RBCs . This destruction of RBCs is called
Haemolysis , and causes Haemolytic anaemia

Metabolism of Lipids
 Lipids are present in our bodies in different forms like triglycerides , phospholipids , fatty
acids,
 The breakdown of lipids to release fatty acids is called lipolysis.
 Triglycerides are (85-90% ) of total lipids , and stored in adipose tissues. They are released
from there to produce energy after long starvation.
ß -Oxidation of Fatty Acids
 ß-Oxidation of fatty acid is a process by which fatty acids are broken down to produce energy .
The beta oxidation of fatty acids occurs in three stages ;
1) Activation of fatty acids in the cytosol.
2) transport of activated fatty acids into mitochondria.
3) Beta oxidation of fatty acid in the matrix of mitochondria.

1. Activation of fatty acids

 Fatty acids are activated to acyl CoA by thiokinase or acyl CoA synthetases. The reaction occurs
in two steps and requires ATP, coenzyme A and Mg2+. Fatty acid reacts with ATP to form
acyladenylate which then combines with coenzyme A to produce acyl CoA. In the activation,
two high energy phosphates are utilized, since ATP is converted to pyrophosphate (PPi). The
enzyme inorganic pyrophosphatase hydrolyses PPi to phosphate (Pi). The immediate
elimination of PPi makes this reaction totally irreversible.
2. Transport of fatty acids into mitochondria
 The inner mitochondrial membrane is impermeable to fatty acids. A specialized carnitine
carrier system (carnitine shuttle) operates to transport activated fatty acids from cytosol to the
mitochondria.

This occurs in four steps

 Acyl group of acyl CoA is transferred to carnitine (β-hydroxy ϒ-trimethyl aminobutyrate),

catalysed by carnitine acyltransferase I (present on the outer surface of inner mitochondrial
membrane).
 The acyl-carnitine is transported across the membrane to mitochondrial matrix by a specific
carrier protein.
 Carnitine acyl transferase II (found on the inner surface of inner mitochondrial membrane)
converts acyl-carnitine to acyl CoA.
 The carnitine released returns to cytosol for reuse

3. β-Oxidation proper in the mitochondrial matrix

 Each cycle of βoxidation, liberating a two-carbon unit-acetyl CoA, occurs in a sequence of four
reactions.
1. Oxidation — Acyl CoA undergoes dehydrogenation by an FADdependent flavoenzyme, acyl
CoA dehydrogenase. A double bond is formed between α and β carbons (i.e., 2 and 3 carbons).
2. Hydration — Enoyl CoA hydratase brings about the hydration of the double bond to form β-
hydroxyacyl CoA.
3. Oxidation — β-Hydroxyacyl CoA dehydrogenase catalyses the second oxidation and generates
NADH. The product formed is β-ketoacyl CoA.
4. Cleavage — The final reaction in β-oxidation is the liberation of a 2- carbon fragment, acetyl
CoA from acyl CoA. This occurs by a thiolytic cleavage catalysed by β-ketoacyl CoA thiolase.

Ketogenesis
 The synthesis of ketone bodies occurs in the liver. The enzymes for ketone body synthesis are
located in the mitochondrial matrix. Ketone bodies are water-soluble and energy yielding.
Acetyl CoA, formed by oxidation of fatty acids, pyruvate or some amino acids, is the precursor
for ketone bodies.
 The three main types of ketone bodies produced are acetone, acetoacetate, and beta-
hydroxybutyrate. Ketone bodies can be used by the brain and other tissues as an alternative
energy source when glucose is scarce, and they are also involved in regulating blood glucose
levels and reducing inflammation.
 However, excessive production of ketone bodies can lead to a condition known as ketoacidosis,
which is a potentially lifethreatening metabolic state characterized by high levels of ketone
bodies in the blood.
Ketogenesis occurs through the following reactions
1. Two moles of acetyl CoA condense to form acetoacetyl CoA. This reaction is catalysed by
thiolase, an enzyme involved in the final step of E-oxidation. Hence, acetoacetate synthesis is
appropriately regarded as the reversal of thiolase reaction of fatty acid oxidation.
2. Acetoacetyl CoA combines with another molecule of acetyl CoA to produce β-hydroxy β-
methyl glutaryl CoA (HMG CoA). HMG CoA synthase, catalysing this reaction, regulates the
synthesis of ketone bodies. 3. HMG CoA lyase cleaves HMG CoA to produce acetoacetate and
acetyl CoA.
3. Acetoacetate can undergo spontaneous decarboxylation to form acetone.
4. Acetoacetate can be reduced by a dehydrogenase to β-hydroxybutyrate.
 Ketolysis
 Ketolysis is the metabolic process by which ketone bodies are broken down and converted into
energy in the body's cells. This process occurs primarily in the mitochondria of cells, where the
ketone bodies are broken down into acetyl-CoA, which can then enter the citric acid cycle to
produce ATP, the energy currency of cells.
 This process is important for individuals who rely on ketone bodies as their primary source of
energy, such as those on a ketogenic diet or during periods of prolonged fasting.
 The rate of Ketolysis is influenced by several factors, including the availability of ketone bodies
and the metabolic state of the cells.
 In some metabolic disorders, such as diabetes, there can be a disruption in the balance
between ketone production and utilization, leading to an accumulation of ketone bodies in the
blood and potentially causing ketoacidosis

Diseases related to abnormal metabolism of lipids

1. Ketoacidosis
 Increase in concentration of both acetoacetate and βhydroxybutyrate (strong acids) in blood would
cause acidosis. The carboxyl group has a pKa around 4. Therefore, the ketone bodies in the blood
dissociate and release H+ ions which lower the PH. Diabetic ketoacidosis is dangerous—may result in
coma, and even death, if not treated. Ketosis due to starvation is not usually accompanied by
ketoacidosis

2. Hypercholesterolemia
 Increase in plasma cholesterol (> 200 mg/dl) concentration is known as hypercholesterolemia
and is observed in many disorders
 Diabetes mellitus Due to increased cholesterol synthesis since the availability of acetyl CoA is
increased.
 Hypothyroidism (myxoedema) This is believed to be due to decrease in the HDL receptors
on hepatocytes.
 Obstructive jaundice Due to an obstruction in the excretion of cholesterol through bile.
 Nephrotic syndrome Increase in plasma globulin concentration is the characteristic feature
of nephrotic syndrome. Cholesterol elevation is due to increase in plasma lipoprotein fractions
in this disorder.

3. Fatty liver
 The normal concentration of lipid in liver is around 5%. Liver is not a storage organ for fat,
unlike adipose tissue. However, in certain conditions, lipids— especially the triacylglycerols—
accumulate excessively in liver, resulting in fatty liver. In the normal liver, Kupffer cells contain
lipids in the form of droplets. In fatty liver, droplets of triacylglycerols are found in the entire
cytoplasm of hepatic cells. This causes impairment in metabolic functions of liver. Fatty liver is
associated with fibrotic changes and cirrhosis,
Fatty liver may occur due to two main causes

 Increased synthesis of triacylglycerols

 Impairment in lipoprotein synthesis

Metabolism Of Amino Acids ( Proteins )

 Proteins are the most abundant organic compounds and constitute a major part of the body
dry weight (10-12 kg in adults).
 They perform a wide variety of static (structural) and dynamic (enzymes, hormones, clotting
factors, receptors etc.) functions. About half of the body protein (predominantly collagen) is
present in the supportive tissue (skeleton and connective) while the other half is intracellular.
 Amino acid—Proteins are nitrogen-containing macromolecules consisting of LD-amino acids
as the repeating units. Of the 20 amino acids found in proteins, half can be synthesized by the
body (non-essential) while the rest have to be provided in the diet (essential amino acids).
 The proteins on degradation (proteolysis) release individual amino acids. Amino acids are not
just the structural components of proteins. Each one of the 20 naturally occurring amino acids
undergoes its own metabolism and performs specific functions.
 Some of the amino acids also serve as precursors for the synthesis of many biologically
important compounds (e.g., melanin, serotonin, creatine etc.).
 Certain amino acids may directly act as neurotransmitters (e.g., glycine aspartate, glutamate).

General Reactions of Amino Acids and Its Significance

General of Amino Acids are following :
1) Transamination : It means movement of an amino (--NH2 ) group from an amino acid to a
Keto Acid . A group of enzymes called Transaminase catalyses this reaction . Keto acid is a
compound that contains a Carboxyl Group ( --COOH ) and a Ketone Group ( >C=O ).
2) Deamination : Loss of amino group from amino acid is Deamination . It is of two types,
Oxidative, Non-Oxidative.
3) Decarboxilation : Removal of CO2 From a compound is called Decarboxilation.
 Transamination
 The transfer of an amino ( NH2) group from an amino acid to a keto acid is known as
transamination. This process involves the interconversion of a pair of amino acids and a pair of
keto acids, catalysed by a group of enzymes called transaminases /aminotransferases.

Silent features of Transamination

 All transaminases require pyridoxal phosphate (PLP), a coenzyme derived from vitamin B6. It
is reversible and no free NH3 liberated, only the transfer of amino group occurs.
Transamination diverts the excess amino acids towards energy generation.
 Specific transaminases exist for each pair of amino and keto acids. However, only two—
namely, aspartate transaminase and alanine transaminase—make a significant contribution for
transamination.
 Transamination is very important for the redistribution of amino groups and production of
non-essential amino acids, as per the requirement of the cell. It involves both catabolism and
anabolism of amino acids.
 The amino acids undergo transamination to finally concentrate nitrogen in glutamate.
Glutamate is the only amino acid that undergoes oxidative deamination to a significant extent
to liberate free NH3 for urea synthesis.
 All amino acids except lysine, threonine, proline and hydroxyproline participate in
transamination. Serum transaminases are important for diagnostic and prognostic purposes.

Mechanism of transamination
 It occurs in two stages
 Transfer of the amino group to the coenzyme pyridoxal phosphate (bound to the
coenzyme) to form pyridoxamine phosphate.
 The amino group of pyridoxamine phosphate is then transferred to a keto acid to
produce a new amino acid and the enzyme with PLP is regenerated.
 Deamination
 The removal of amino group from the amino acids as NH3 is deamination. Deamination results
in the liberation of ammonia for urea synthesis (transamination involves only the shuffling of
amino groups). It may be either oxidative or non-oxidative.

1. Oxidative deamination : Oxidative deamination is the liberation of free ammonia from the amino
group of amino acids coupled with oxidation. This takes place mostly in liver and kidney. The purpose
of oxidative deamination is to provide NH3 for urea synthesis and D-keto acids for a variety of
reactions, including energy generation

 Oxidation of glutamate-by-glutamate dehydrogenase : In the transamination process glutamate is

form. Now, Glutamate rapidly undergoes oxidative deamination, catalysed by glutamate dehydrogenase
(GDH) to liberate ammonia. This enzyme is unique in that it can utilize either NAD+ or NADP+ as a
coenzyme. Conversion of glutamate to α-ketoglutarate occurs through the formation of an
intermediate, α-iminoglutarate.
 Oxidative deamination by amino acid oxidases : L-Amino acid oxidase and D-amino acid oxidase
are flavoproteins, possessing FMN and FAD, respectively. They act on the corresponding amino acids (L
or D) to produce D-keto acids and NH3. In this reaction, oxygen is reduced to H2O2, which is later
decomposed by catalase.

2. Non-oxidative deamination : Some of the amino acids can be deaminated to liberate NH3
without undergoing oxidation.

a. Amino acid dehydrases : Serine, threonine and homoserine are the hydroxy amino acids.
They undergo non-oxidative deamination catalysed by PLP-dependent dehydrases
(dehydratases).
b. Amino acid desulfhydrases : The sulphur amino acids, namely cysteine and homocysteine,
undergo deamination coupled with desulfhydration to give keto acids.
c. Deamination of histidine : The enzyme histidase acts on histidine to liberate NH3 by a non-
oxidative deamination process
 Urea cycle
 Ammonia is constantly being liberated in the metabolism of amino acids (mostly) and other
nitrogenous compounds. At the physiological pH, ammonia exists as ammonium (NH4+) ion.
Ammonium ions are very important to maintain acid-base balance of the body.
 The production of NH3 occurs from the amino acids (transamination and deamination),
biogenic amines, amino group of purines and pyrimidines and by the action of intestinal
bacteria (urease) on urea.

Introduction about urea cycle

 Urea is the end product of protein metabolism (amino acid metabolism). The nitrogen of
amino acids, converted to ammonia, is toxic to the body. It is converted to urea and detoxified.
As such, urea accounts for 80-90% of the nitrogen containing substances excreted in urine.
 Urea is synthesized in liver and transported to kidneys for excretion in urine. Urea cycle is the
first metabolic cycle that was elucidated by Hans Krebs and Kurt Henseleit (1932), hence it is
known as Krebs-Henseleit cycle. The individual reactions, however, were described in more
detail later on by Ratner and Cohen. Urea has two amino (NH2) groups, one derived from NH3
and the other from aspartate. Carbon atom is supplied by CO2. Urea synthesis is a five-step
cyclic process, with five distinct enzymes. The first two enzymes are present in mitochondria
while the rest are localized in cytosol
 Decarboxylation
 Decarboxylation of an amino acid is a chemical reaction in which the carboxyl group (-COOH)
of an amino acid is removed, leading to the formation of an amine group (-NH2) and the
release of carbon dioxide (CO2). This reaction is catalysed by enzymes called decarboxylases.
 In general, the decarboxylation of amino acids or their derivatives results in the formation of
amines.
 The decarboxylation of amino acids plays an important role in many biological processes. For
example, the amino acid histidine is decarboxylated to form histamine, which is involved in
regulating many physiological processes, including digestion and immune response. Similarly,
the amino acid glutamic acid is decarboxylated to form the neurotransmitter gamma-
aminobutyric acid (GABA), which is involved in the regulation of neuronal activity.

Significance of amino acid metabolism

 It is necessary for proper functioning of body .
 It is very important pharmacologically also .
 Some are important precursor of steroid hormones .
 Some function as Coenzyme .
 Some are act as neurotransmitter ( GABA)
Diseases Related to Abnormal Metabolism of Amino Acids
1) Albinism : It is a Group of inherited disorder it develops when tyrosine metabolism does not occur
properly , and then synthesis of melanin becomes very low or no melanin production . Melanin is a
pigment , so the affected person has little or no colour in hair and skin . this condition increases the
risk of skin cancer

2) Tyrosinemia : It is a inherited disorder in which metabolism of t yrosin badly affected and

accumulate in body tissues , and because of its toxic effect many of problems created .

Symptoms :

 Bloody stool
 Diarrhoea
 Fatigue
 Vomiting.
 Painful wounds on skin,
 Red eyes
 convulsion
 intelactual
 Intellectual disability.

Disorder of Ammonia Metabolism

1) Phenylketonuria (PKU) : It is a Congenital disorder characterized by inability to convert
phenylalanine to tyrosine , so phenylalanine accumulated in the body and appears in urine

Symptoms :

 Delay in growth and development

 Mental problem ( retardation )
 Psychiatric disorder
 Weakness of bone
 Rashes on skin
 Fair skin and blue eyes
 Hyperactivity and behavioral problems.

2) Alkaptonuria ( black Urine Disease ) It is a rare and genetic disorder , in which Homogentisic
Acid accumulated in the tissues due to low production of HOMo- Gentisic - Dioxygenase enzyme .

Symptoms : Dark or black coloured urine upon air exposure.

3) Jaundice

 Amino acid metabolism disorders can lead to jaundice if there is a disruption in the pathway
that converts bilirubin into a form that can be eliminated from the body. For example, a
genetic disorder called CriglerNajjar syndrome can lead to jaundice because it impairs the
ability of the liver to convert bilirubin into a form that can be excreted in the bile. This can
cause bilirubin to build up in the blood, leading to jaundice.
 Similarly, other genetic disorders such as Gilbert's syndrome can cause jaundice due to
problems with bilirubin metabolism. In Gilbert's syndrome, there is a deficiency of an enzyme
called UDPglucuronosyltransferase, which is responsible for conjugating bilirubin so that it can
be eliminated from the body. Without this enzyme, bilirubin builds up in the blood and can
lead to jaundice

Biological Oxidation
 Oxidation is a reaction with oxygen directly or indirectly / removal of hydrogen or electron .
 This reaction carried out by enzymes .
 The electron released by this reaction accepted by Electron acceptors ( NAD , FAD ) , and then
formation of ATP occurs , this process takes place in living tissues , and necessary for survival
so it is called Biological Oxidation .

Electron Transport Chain and Oxidative Phosphorylation

 Electron Transport Chain is a series of protein complex and other molecules that accept and
transfers electron from NADH and FADH2 to Oxygen , when they combine with oxygen the
synthesis of ATP occurs.
 In formation of ATP Phosphorus is used and Oxidation - Reduction reaction involved that is
why it called Oxidative phosphorylation.
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