Biochemistry Part01 R01

BIOCHEMISTRY
A COMPLETE COURSE
PART I
DR. C. SHANMUGAPRIYA
PROFESSOR OF BIOCHEMISTRY
Dr. C. Shanmugapriya,
MBBS., M.D., (Biochemistry)
saipriyain@gmail.com
She is working as the Head, Research & Development (May 2018
till date) of KRIYA Medical Technologies Pvt. Ltd. She has been instru-
mental in products validation and in new products development includ-
ing a unique molecular transport media for SARS CoV2, molecular diag-
nostic kit for Omicron detection and for simultaneous detection of SARS
CoV2, Influenza A & B, RSV and is currently undergoing training in novel
Molecular Biology Techniques at KRIYA.
She has done her MBBS at Madurai Medical College and MD Bio-
chemistry at Madras Medical College. She was working as the Professor
and Head of the Department of Biochemustry, Government Thoothukudi
Medical College and Hospital, Thoothukudi. She has been teaching Bio-
chemistry and Physiology for medical and dental post-graduation aspir-
ants since 2006 and she has coached thousands of students for the past
16 years, who continue to come up with flying colours. She is known for
her conceptual teaching. She is the co-author of the book “A guide for
preparation of ALL INDIA NEET EXAM” Volume I 2016-2017.
She was representing India in the International Federation of Clini-
cal Chemistry – Task Force for Young Scientists for three consecutive
terms 2012 to 2015, 2015 to 2018, and 2019 to 2021. She has received
the “Young scientist award” twice, in the year 2010 for the work on Type
2 diabetes and in the year 2008 for the work on coronary atheroscle-
rosis. Her work on Type 2 diabetes was also chosen as the best scien-
tific paper in the National Diabetology Conference in the year 2010 at
Chennai and she received the Best oral paper presentation for her
work on “TCF7L2 Variant rs7903146 affects the Risk of Type 2 Diabetes
by Modulating Incretin secretion / action” at Asia Pacific Federation of
Clinical Biochemists 2013 held at Bali.
She is an educator in Prepladder, an online teaching platform for
medical postgraduation aspirants since March 2023 and she will be
teaching Biochemistry and Applied Biochemistry in the platform.
She is an educator in Unacademy in the NEET PG category where she
teaches Biochemistry and Physiology to NEET PG aspirants.
https://unacademy.com/@CShanmugapriya
She has a YouTube channel “Biochemistry Dr.C. Shanmugapriya” in which
she uploads conceptual videos on Biochemistry and Clinical Biochem-
istry.
https://www.youtube.com/channel/UCTge8U1HBZf7jUaBATr1iCw
For more details and more testimonials, check her page:
https://www.facebook.com/biochemneetpgdiscussionforum/
@dr.c.shanmugapriya Dr. C. Shanmugapriya BIOCHEMISTRY
Contents
Introduction 05 Non protein part of an enzyme 91
BIOCHEMISTRY I A COMPLETE COURSE I Part 01
Carbohydrate Enzyme Kinetics and Enzyme

Chemistry Part 1 13 Inhibition 92
Algorithm for reducing sugars in Electron Transport Chain 100
urine 19 Introduction 101
Carbohydrate Chemistry Part Shuttles 101
2 23
Mitchelle’s Chemiosmotic theory
Mucopolysaccharides 24 103
Isomerism 27 Inhibitors 106
Carbohydrate metabolism 34 Heme Synthesis and
Introduction 35 Porphyrias 110
Glycolysis 39 Introduction 111
Fates of Pyruvate 51 Heme synthesis 112
Pyruvate Dehydrogenase 53 Heme synthesis disorders 115
Tca cycle 57 Lipid Chemistry 118
Glycogen metabolism 61 Classification 119
Glycogenolysis 63 Glycolipds 126
Glycogen Storage Disorders 69 Classification of lipids based on
Gluconeogenesis 75 solubility 131
Enzymes 86 Fatty acids 132
Introduction 87 Lipid metabolism 134
Clasification of Enzymes 87 Fatty acid synthesis 135
4 >>>
Introduction
Pathways and Suborganelles
Pathways Suborganelles
Glycolysis >
PDH >
TCA Cycle >
Glycogen metabolism >
Gluconeogenesis >
HMP SHUNT >
Fatty Acid Synthesis >
Fatty Acid Oxidation >
Ketone body synthesis >
Cholesterol synthesis >
Bile acid Synthesis >
Urea Cycle >
Heme synthesis >
6 >>>
Pathways Suborganelles
Glycolysis > Cytoplasm
PDH > Mitochondria
TCA Cycle > Mitochondria
Glycogen metabolism > Cytoplasm
Gluconeogenesis > Mitochondria,

Cytoplasm and ER
HMP SHUNT > Cytoplasm
Fatty Acid Synthesis > Cytoplasm
Fatty Acid Oxidation > Mitochondria and

Peroxisome
Cholesterol synthesis > Cytoplasm and ER
Ketone body synthesis > Mitochondria
Bile acid Synthesis > ER
Urea Cycle > Mitochondria and

Cytoplasm
Heme synthesis > Mitochondria and

Cytoplasm
Other points to remember: Suborganelles
• Ether lipid synthesis

• The only oxidation which is anaerobic is
glycolysis and hence glycolysis is the only • Bile acid conjugation
pathway that happens in cytoplasm • Rough endoplasmic reticulum is related to
Fatty acid oxidation happens in mitochon- protein synthesis and smooth endoplasmic
dria but very long chain fatty acids get reticulum (SER) is related to steroid synthe-
oxidised in peroxisomes sis. Hence cholesterol synthesis and bile
acid synthesis happen in SER
• Functions of peroxisomes:
• Very long chain fatty acid oxidation
• The two pathways which happen in both
cytoplasm and mitochondria are Urea cycle
• Branched chain fatty acid oxidation
and heme synthesis
7
Marker enzymes
S.No Organelle Marker enzyme
1 Nucleus >
2 Endoplasmic reticulum >

3 Golgi complex >
4 Mitochondria >
Outer
membrane
>
Inner
Membrane
5 Lysosome >
6 Cytoplasm >
7 Peroxisome >
8 >>>
S.No Organelle Marker enzyme
1 Nucleus > DNA polymerase/

RNA Polymerase
2 Endoplasmic reticulum > Glucose 6

Phosphatase
3 Golgi complex > Galactosyl

transferase
4 Mitochondria Outer > Monoamine Oxidase

membrane
Inner > ATP synthase

Membrane
5 Lysosome > Cathepsin
6 Cytoplasm > Lactate

Dehydrogenase
7 Peroxisome > Catalase
Other points to remember: Marker enzyme
• Replication and Transcription occur in • Marker enzykme of Outer Membrane

Nucleus. Hence DNA and RNA polymer- of Mitochondria is Monoamine Oxidase
ases are the marker enzymes for nucleus • Electron transport chain is present
• Glucose 6 phosphatase is an enzyme along the inner side of inner mitochod-
of gluconeogenesis and is present in En- nrial membrane, hence for the inner
doplasmic reticulum, Hence the marker membrane of mitochondria, the marker
enzyme for ER or microsome is Glucose enzyme should be one of the complexes
6 phosphatase of ETC – Complex II or Succinate dehy-
• Golgi complex is related to glycopro- drogenase and Complex V or ATP syn-
tein synthesis. The enzyme involved thase
in Glycoprotein synthesis is galactosyl • Hydrogen peroxide is generated Per-

transferase. Hence the marker enzyme oxisome is detoxified by catalase. Hence
for Golgi Complex is Galactosyl Trans- catalase is the marker enzyme for perox-
ferase isome
9
Where are proteins synthesised?

• Proteins are synthesised by free ribo- es can be attached only by Endoplasmic
somes. Ribosomes read the mRNA from 5’ reticulum and by Golgi complex, Rough En-
end to 3’ end. They read every codon one doplasmic reticulum is formed. (the target-
by one. Depending upon the codon that ing sequence required for lysosomal protein
is present, they recruit a complementary is mannose 6 phosphate and for membrane
anticodon containing tRNA. tRNA carries an protein is a lipid side chain. If a protein
aminoacid. This is how nucleotide sequenc- carries neither a mannose 6 phosphate
es of mRNA get translated to aminoacid residue nor a lipid side chain, it gets target-
sequences of polypeptide chain. This way ed as a secretory protein) In this case, free
aminoacids get synthesised from the ami- ribosomes initiate the synthesis of proteins
noterminal end to the carboxy terminal end and then, the amino terminal end of these 3
and the protein synthesis gets over if the types of proteins have a signal recognition
protein that is getting synthesised is a nu- peptide, which guides the ribosomes to get
clear protein or a cytoplasmic protein or a attached to Endoplasmic reticulum to form
mitochondrial protein. Rough Endoplasmic reticulum.
• If the protein that is getting synthesised • If a modification happens within rough
is cytoplasmic or membrane or secretory endoplasmic reticulum, it is called as a
protein, because we want specific targeting cotranslational modification. If a modification
sequences to target them to the respective happens in golgi complex or thereafter, it is
organelles, and as these targeting sequenc- a post translational modification.
10 >>>
I cell disease
• It is a lysosomal storage disorder caused • Lysosomes are empty bags and hence is
by the defect of N-acetylglucosamine phos- characterized by inclusions, hepatospleno-
photransferase megaly and coarse facial features
• It is a protein targeting defect • Serum lysosomal enzyme activity is high
• Lysosomal enzymes are not targeted to • Tissue lysosomal enzyme activity is high
lysosomes but are secreted as secretory
proteins
Hyperoxaluria
It is characterised by recurrent renal stones. aminotransferase being placed in mitochon-
It is of two types: dria instead of being placed in peroxisomes.
>> Primary hyperoxaluria >> Type II – It is caused by a defect of Gly-
>> Secondary Hyperoxaluria oxolate reductase
Primary Hyperoxaluria Secondary Hyperoxaluria
>> It is caused by a defect of glycine catab- >> Intake of oxalate rich food – chocolate,
olism. tea, beetroot, green leafy vegetables
>> Type I – It is a protein targeting defect >> Enteric hyperoxaluria – 5 to 10% of
caused by the enzyme Glyoxalate alanine oxalate in the diet is absorbed – insoluble
calcium oxalate is excreted in feces
• Primary hyperoxaluria
• Glycine metabolism
PEROXISOME
OXALATE
GLYCINE GLYOXALATE
GLYOXALATE GLYCHOLATE
ALANINE AMINO
GLYOXALATE
TRANSFERASE
REDUCTASE
PYRUVATE
ALANINE
11
MCQS
1. Which of the following pathways only
suspects a LSD and asks the IEM lab to
perform few lysosomal enzyme activities
takes place in a cell’s cytoplasm? including hexosaminidase A. All lysosomal
enzyme activities were high. What is the
A. Glycolysis probable diagnosis?
B. Beta oxidation
C. TCA A. Tay sach’s disease
D. Urea cycle B. Mucopolysaccharidosis I
2. The marker enzyme of microsomes is

C. Mucopolysaccharidosis II
D. I cell disease
A. Galactosyl transferase 6. A 39 year old man came to the emer-

B. Cathepsin gency department because of severe back
C. Lactate dehydrogenase pain. An ultrasound was taken and renal
D. Glucose 6 phosphatase stones were found. Following lithotripsy a
3. Cytoplasmic proteins are synthe-

sample of the stone was sent for biochemi-
cal analysis. Results revealed presence of ox-
sized in? alate and glyoxalate. He was diagnosed with
primary hyperoxaluria Type I. It is caused by?
A. Ribosomes
B. Nucleus A. Protein folding defect
C. Smooth Endoplasmic reticulum B. Silent mutation
D. Rough endoplasmic reticulum C. Protein targeting defectd.
4.Secretory proteins are synthesized in?

D. Acceptable mutation
A. Ribosomes
B. Smooth Endoplasmic reticulum
Answers: 1.A; 2.D; 3.A; 4.D; 5.D; 6.C.
C. Rough endoplasmic reticulum

D. First in ribosomes and then in endoplas-
mic reticulum
5. A child presents with coarse features,

HSM and mental retardation. The clinician
12 >>>
Carbohydrate
Chemistry
PART I
Carbohydrates
types – Glycoproteins, Proteoglycans

• Carbohydrates are polyhydroxy aldoses
and Glycolipids
or ketoses having the general molecular
formula CnH2nOn • Simple carbohydrates:
• Carbohydrates are classified as: >>Depending on the number of carbohy-
drate units present, simple carbohydrates
• Simple carbohydrates – They have only car-
are classified as
bohydrate units in them
• Monosaccharides – one sugar unit
• Complex carbohydrates – They have in
addition to carbohydrate residues either • Oligosaccharides – two to 10 sugar units
lipids or proteins attached. According- • Polysaccharide – 11 or more number of
ly complex carbohydrates are of three sugar units
Comparison of glucose & fructose
Monosaccharide Monosaccharide
Aldose Ketose
Hexose Hexose
GLUCOSE FRUCTOSE
14 >>>
Classification of carbohydrates
MONOSACCHARIDES
SIMPLE OLIGOSACCHARIDES
POLYSACCHARIDES
CARBOHYDRATES
GLYCOPROTEIN
COMPLEX PROTEOGLYCAN
GLYCOLIPID
Monosaccharides
• Pentoses: They have 5 carbon atoms
• Monosaccharides are classified in
two ways: • Hexoses: They have 6 carbon atoms and
so on
• Based on the number of carbon atoms,
they are classified as • Based on the functional group that is
present, they are classified as
• Trioses: They have 3 carbon atoms (mis-
nomer is maltotriose which has 3 glu- • Aldoses: Have an aldehyde group
cose residues with 18 carbon atoms) • Ketoses: Have a ketone group – ketoses
• Tetroses: They have 4 carbon atoms are named with a “ul”
Classification of monosaccharides
TRIOSES Exception:
NUMBER OF Maltotriose –
TETROSES (glu)3
CARBON ATOMS
PENTOSES
MONOSACCHARIDES
HEXOSES
FUNCTIONAL
ALDOSES
GROUP
KETOSES Ketose – “ul”
15
Disaccharides
• Disaccharides have two sugar units. • The individual sugars and linkages pres-
ent in all disaccharides are as follows
S.No Disaccharide Sugar Linkage
1 Maltose Glu + Glu α (1,4)
2 Isomaltose Glu + Glu α (1,6)
3 Trehalose Glu + Glu α (1,1)
4 Sucrose Glu + Fru α (1,2)

5 Lactose Gal + Glu β (1,4)
Facts to be known about linkages

• Sucrose has α (1,2) and hence it is non
• All carbohydrates have α (1,4) along strai-
reducing
ght chains and α (1,6) along branch points
• Lactose has β (1,4) linkage – clue –
• Exceptions:
lactase is the only human digestive
• Isomaltose has α (1,6)
enzyme which can act on β linkage
• Trehalose has α (1,1)
Exception: Sucrose with α (1,2) linkage
Sucrose is
non reducing α (1,2)
because the
linkage present
in sucrose is
α (1,2)
GLUCOSE FRUCTOSE
16 >>>
Lactose intolerance
• Caused by lactase deficiency in the • Undigested lactose is utilised by intes-
intestinal villi tinal microorganisms to form Hydrogen
• Undigested lactose can not be absorbed and methane
• Lactose stays in the intestinal lumen, • Hydrogen and methane are the reason
attracting water. for bloating, flatulence and frothy stools
• This causes osmotic diarrhoea • Undigested lactose are used by some
microorganisms to form acids – the ex-
planation for acidic pH of stools
Investigations for lactose intolerance

>> Hydrogen or methane breath test • If hydrogen in the breath is increased
• After overnight fast, baseline breath by more than 20ppm than the previ-
hydrogen is measured. 20 to 25 g of ously measured value during the test
lactose is provided. period, it is positive
• Hydrogen in the breath is measured at >> Stool acidity test

15, 30, 60 min and 2 hours • For neonates
Reducing and non reducing sugar

Carbohydrates based on the presence or absence of
reduction property are classified as:
1. Reducing sugars: Any carbohy- 2. Non reducing sugars: Sucrose,

drate with a free carbonyl group is reducing trehalose, oligosaccharides and polysac-
eg., all monosaccharides and most of the charides
disaccharides, except sucrose and trehalose
REDUCING Monosaccharides
SUGARS Most of the
disaccharides
CARBOHYDRATES
NON Sucrose
REDUCING Trehalose
SUGARS Oligosaccharides
Polysaccharides
17
Benedict’s test
• Used to differentiate reducing and non
reducing sugars.
• To 5 mL of benedict’s solution, we add
8 drops of the solution to be tested and is
heated.
• If there is a reducing substance, there is
a color change from blue to green or yel-
low or orange or red, depending upon the
concentration of the reducing substance
in the given solution. Hence the test is
called as a semiquantitative test.
Components of benedict’s solution
>> Copper sulphate: pro- >> Sodium Carbonate: >> Sodium Citrate: stabilis.
vides the blue color. provides the alkaline medium es the solution.
Barfoed’s test
provides an acidic medium, which is
• Barfoed’ reagent composition
unfavorable
• Copper acetate – provides the blue
• Hence, Benedict’s test is answered
color
by monosaccharides and reducing
• Glacial acetic acid– provides an acid-
disaccharides whereas Barfoed’s test
ic medium
is answered by only strongly reducing
• Difference between Benedict’s and Bar-
monosaccharides.
foed’s test
• Hence, Barfoed’s test is used to
• Benedict’s provides an alkaline medi-
differentiate monosaccharides from
um, which is favorable for reduction
reducing disaccharides
property whereas Barfoed’s reagent
Benedict’s test positive in urine -

interpretation
disaccharide – the only disaccharide that
• Presence of reducing sugars in urine eg,
can be present in urine is lactose as it is the
monosaccharides like glucose or fructose or
only one that can be synthesised in body
galactose or pentose or lactose a reducing
18 >>>
• Phsyiologically excreted substances

• Presence of non carbohydrate reducing
when found in excess – eg., uric acid
substances like
or homogentisic acid in alkaptonuria
• Antioxidants like Vitamin A,C,E and
• Drugs which get metabolised by glucu-
Glutathione
ronidation eg., Aspirin
Benedict’s test
Positive Negative
Monosaccharides Oligosaccharides
Sucrose,Trehalose
Most Disaccharides Polysaccharides
Antioxidants – Vitamin A, C, E & GSH
Uric Acid
Homogentisic acid
Drugs metabolised by
glucuronidation
Algorithm for reducing

sugars in urine
When Benedict’s test is positive
Step 1: Take history to rule out drug intake lactation from other causes, Barfoed’s test is
and to rule out clinical features of Alkap- performed. If Barfoed’s test is negative it is a
tonuria disaccharide – lactose, if its positive, it is one
of the monosaccharides
Step 2: Rule out a physiological cause
– Pregnancy and lactation, in which case Step 4: To identify the specific monosac-
lactose is found in urine. Lactose is the only charides, the following tests can be per-
reducing disaccharide that can be found in formed
urine • Glucose – Glucose Oxidase and Peroxi-
Step 3: Find out if its glucose or fructose or dase method
galactose or pentose, all of these are mon- • Fructose – Seliwanoff test or Foulger’s test
osaccharides. Hence, to differentiate the • Galactose – Mucic Acid test
physiological conditions like pregnancy or • Pentose – Bial’s test
19
BENEDICT’S EXCLUDE
TEST antioxidant
intake, drugs,
Alkaptnuria
Physiological HISTORY
Pathological
causes causes
Lactose
Pregnancy &
Lactation Glucose Fructose Galactose Pentose
Diabetes
Essential
Mellitus Classical
fructosuria Essential
Renal Glycosuria Galactosemia
Fructose Pentosuria
Alimentary Galactosemia
Intolerance
glycosuria
BENEDICT’S
TEST
- BARFOED’S +
Physiological TEST
causes
Pathological
causes
Lactose
Pregnancy &
Lactation Glucose Fructose Galactose Pentose
GOD/POD Seliwanoff Mucic Acid

Bial’s test
TEST Foulger test
Polysaccharides
• Structural homopolysaccharides e.g.,
• Polysaccharides have 11 or more num-
cellulose, inulin, chitin. Cellulose is a
ber of sugar units. They are of two types:
homopolysaccharide made up of glu-
>> Homopolysaccharides: The individual
cose and is linked by Beta (1,4) linkage.
units are the same. They are classified into
It is a component of plant cell wall. Inulin
two types
is a homopolysaccharide made up of
20
fructose. Chitin is a homopolysaccharide >> Heteropolysaccharides: The individ-

made up of NAG ual units are different eg., Mucopolysaccha-
• Storage homopolysaccharides – rides of Glycosaminoglycans
Branched eg., Starch and glycogen
Starch
Starch has two structures within itself • Amylopectin – Branched with multiple
glucose residues connected by alpha
• Amylose – unbranched with multiple
(1,4) linkages along straight chains and
glucose residues connected by alpha
alpha (1,6) linkages at branch points. They
(1,4) linkages. They are not that stable and
are stable and hence the melting temper-
hence the melting temperature is low. At
ature is high. At room temperature it is yet
room temperature, it is already melted and
to melt and hence accounts for the solid
so accounts for the liquid part of starch
part of starch
Glycogen
• It is stored in Liver and skeletal muscle • At branch points there are α (1,6) linkages
• Glycogen is the most branched structure • The entire structure is arranged in 12
ever known concentric layers
• Centre of glycogen has a protein called as • Branch points are found in the inner layer
glycogenin and unbranched in the outer layers
• Every straight chain has 11 to 13 glucose
residues linked with α (1,4) linkages
MCQS
1. Which of the following 2. All the following are 3. The linkage present in
is a ketose? true about glucose except? lactose is:
A. Arabinose A. It is a monosaccharide A. α(1,4)

B. Erythrose B. It is an aldose B. β(1,4)
C. Ribose C. It is a hexose C. α(1,2)
D. Sedoheptulose D. It is non reducing D. β (1,2)
21
4.The linkage present in isomaltose is: 8. All the following are component of
Benedict’s solution except?
A. α (1,4)
B. β(1,4) A. Copper sulphate
C. α (1,6) B. Sodium Carbonate
D. β (1,6) C. Sodium Citrate
5. A 21 year old man presents with diar-

D. Acetic acid
rhea, bloating, flatulence and frothy stools 9. All the following answer Benedict’s
every time he consumes milk, ghee and test positively except?
butter. Mention the enzyme that is deficient
in this person. A. Ribose
B. Vitamin C
A. Aldolase B C. Homogentisic acid
B. Galactose 1 phosphate uridyl trans- D. Trehalose
10. All of the following are branched

ferase
C. Lactase
D. Fructokinase except
6. A 21 year old man presents with diar- A. Amylose

rhea, bloating, flatulence and frothy stools B. Amylopectin
every time he consumes milk, ghee and but- C. Glycogen
ter. He is suspected to suffer from lactose D. Starch
6- A: VITAMIN B12 MALABSORPTION; B: FOLATE DEFICIENCY; PHENYL KETONURIA

11. A neonate presented with hypogly-
intolerance. What is the investigation you will
ask for?
cemia. The pediatrician performed a bed
A. Schilling’s test side urine test to narrow down his provi-
B. FIGLU test sional diagnosis. Urine Benedict’s test was
Answers: 1.D; 2.D; 3.B; 4.C; 5.C; 6.D. 7.D; 8.D; 9.D; 10.A;11.C.
C. Guthrie’s test positive. He sent the urine sample to a
D. Hydrogen breath test clinical laboratory for further investigations.
7. Which of the following is reducing?

The lab provided the following results:
I. Benedict’s test : Positive
II. Barfoed’s test : Positive
A. Sucrose III. GOD POD test : Negative
B. Starch IV. Seliwanoff test : Positive
C. Glycogen The substance present in the person’s
D. Lactose urine is
A. Glucose
B. Homogentisic acid
C. Fructose
D. Galactose
22 >>>
Carbohydrate
Chemistry
PART II
MUCOPOLYSACCHARIDES
& ISOMERISM
Mucopolysaccharides
MPS/GAG
ride made up of repetitive units of galac-
• Mucopolysaccharides or Glcosaminogly-
tose and aminosugar
can is a heteropolysaccharide
• Iduronic acid is present in Heparin, Hepa-
• It is made up of long, straight unbranched ran sulphate and Dermatan Sulphate
chains with Uronic acid and Aminosugar.

• Galactosamine is present in Chondroitin
Some of these residues are additionally sul-
sulphate and Dermatan Sulphate
phated and/or acetylated
• Hyaluronic acid is neither sulphated or
• The thumb rule is that all mucopolysac- acetylated
charides are made up of glucuronic acid
and glucosamine
• The mucopolysaccharide which follows
all thumb rules is Hyaluronic acid
• Exceptions:
• The mucopolysaccharide which does
• Without uronic acid is Keratan Sulphate.
not follow both the exceptions is Der-
Keratan sulphate is a mucopolysaccha-
matan Sulphate
Tips to identify the structure
If it is below the plane of the ring, it is idu-

• Identify acid part and aminosugar part
ronic acid
• If no acid, it is keratan sulphate
• In the aminosugar part, check the OH
• In the acid part, check the location of the group in the fourth carbon atom. If OH is
carboxyl group (6th carbon atom). If it is above the plane, it is galactosamine. If it is
above the plane of the ring, it is uronic acid. below, it is glucosamine.
24 >>>
Identify the structure
HYALURONIC
6 6 ACID
5 5
4 4 1
1
2 2
3 3
GLUCURONIC ACID GLUCOSAMINE
CHONDROITIN
SULPHATE
GLUCURONIC ACID GALACTOSAMINE
DERMATAN
SULPHATE
IDURONIC ACID GALACTOSAMINE
25
KERATAN
SULPHATE
NO URONIC ACID
Location of MPS
S.No Mucopolysaccharide Location
1 >
2 >
3 >
4 >
5 >
6 >
7 >
26 >>>
S.No Mucopolysaccharide Location
1 Hyaluronic acid > Synovium & Vitreous
2 Keratan Sulphate I > Cornea
Keratan Sulphate II > Loose connective

3 tissues
4 Heparin > Mast cells
5 Heparan sulphate > Aorta, GBM
6 Chondroitin sulphate > Cartilages and

Bones
7 Dermatan sulphate > Universal
Isomerism
• Isomerism is a property by which two or form. If an isomer turns it to the left side it is
molecules have the same molecular formula levorotatory or l or – form
but they have different structural formula >> Functional Isomerism: Here the two
or they differ in spatial orientation of their molecules have the same molecular formula
groups but different functional groups. Eg., aldoses
• It is of three types and ketoses are functional isomers- glucose
and fructose
• Optical isomerism
• Functional isomerism >> Stereoisomerism: The two molecules
• Structural Isomerism have the same molecular and structural
formula but they differ in spatial prienta-
>> Optical Isomerism: It is a property by
tion of their groups. It is dependent on the
which two or more molecules have the same
presence of an asymmetric carbon atom.
molecular formula but they differ in the way
The number of stereoisomers in a molecule
they turn the plane polarised light. If an iso-
containing n number of carbon atoms is 2n.
mer turns the plane polarised light towards
The number of stereoisomers possible for
the right side, it is dextrorotatory or d or +
glucose is 32 and for fructose is 16.
27
Stereoisomerism
Stereoisomerism is of four types
> Anomerism
> Epimerism
> Diastereoisomerism
> Enantiomerism
• Anomerism: It is a property by which two or more mol-

ecules have the same molecular formula and the same
structural formula, but they differ in spatial orientation in the
anomeric carbon atoms or in the functional carbon atom. In
the functional carbon atom, if the OH group lies below the

functional carbon atom it is called as alpha anomer, if the
OH group lies above the functional carbon atom, it is called
as beta anomer
• Epimerism: It is a property by which two or more mole-

cules have the same molecular formula and the same struc-
tural formula, but they differ in spatial orientation in one of
the asymmetric carbon atoms other than the penultimate
carbon atom.
• Example glucose and mannose are C2 epimers, glucose
and galactose are C4 epimers. Ribose and arabinose are C2
epimers, Ribose and Xylose are C3 epimers
• Diastereoisomerism: It is a property by which two or
more molecules have the same molecular formula and the
same structural formula, but they differ in spatial orientation
in one or more of the asymmetric carbon atoms other than
the penultimate carbon atom.
28 >>>
• Example galactose and mannose are fer in spatial orientation in all the asymmetric
diastereoisomers, Xylose and arabinose are carbon atoms. They are named on the basis
diastereoisomers of the orientation in the penultimate carbon
• Enantiomerism: Enantio is mirror. Enan- atom. In the penultimate carbon atom, if OH
tiomerism is a property by which the two is on the right time, it is called as D form, if
molecules have the same molecular formula its on the left time, it is called L form. The
and the same structural formula and they dif- other name for enantiomerism is racemism
Epimers of glucose
GLUCOSE GALACTOSE
Epimers of ribose
RIBOSE ARABINOSE XYLOSE
29
Closed ring form of glucose

GLUCOSE
ISOMERISM
OPTICAL FUNCTIONAL
STEREOISOMERISM
ISOMERISM ISOMERISM
Aldoses and Glucose and

d and l form
ketoses Fructose
ISOMERISM
OPTICAL FUNCTIONAL
STEREOISOMERISM
ISOMERISM ISOMERISM
ENANTIOMERISM DIASTEREOMERISM EPIMERISM ANOMERISM
1. Galactose & 1. Glucose &

Mannose Mannose
D&L Beta & Alpha
2. Arabinoase & 2. Glucose &
Xylose Galactose
30 >>>
MCQS
1. All the following are components of 5. The mucopolysaccharide present in
Mucopolysaccahrides except? glomerular basement membrane is?
A. Uronic acid A. Hyaluronic acid

B. Aminosugar B. Heparan sulphate
C. Sulphate C. Dermatan sulphate
D. NANA D. Keratan sulphate
6. A 18 month old female child presented

2. The mucopolysaccharide with galac- with flat nasal bridge, big lips, macroglossia,
tose is? gingival hypertrophy, abdominal distension,
short stature, delayed milestones. On exami-
A. Hyaluronic acid nation, hepatosplenomegaly and an inguinal
B. Heparin hernia was observed. Chest X ray shows car-
C. Heparan sulphate diomegaly. Urinary MPS was elevated. WBC
D. Keratan sulphate L-iduronidase activity was low. Which of the
3. The mucopolysaccharide with glucu-

following would accumulate?
ronic acid is? A. Heparan sulphate

B. Keratan sulphate
A. Hyaluronic acid C. Hyaluronic acid
B. Heparin D. Chondroitin sulphate
C. Dermatan sulphate
D. Keratan sulphate
4. The mucopolysaccharide with glu-

cosamine is?
A. Hyaluronic acid
B. Chondroitin sulphate
C. Dermatan sulphate
D. Keratan sulphate
31
S.No PROPERTY HURLER’S SYNDROME HUNTER’S SYNDROME
3
S.No PROPERTY HURLER’S SYNDROME HUNTER’S SYNDROME
MPS type I II
2 Enzyme L- IDURONIDASE L-IDURONATE

defect SULPHATASE
Severity Severe Mild
4 Clinical Coarse facial features, No Inguinal hernia and

features Gingival hypertrophy, no corneal clouding
depressed nasal
bridge, short stature,
HSM, Inguinal hernia,
Cardiomegaly, Intellectual
disability, Retinal
degeneration, corneal
5 Inheritance Autosomal recessive X linked recessive
32 >>>
KEY
>> Hurler syndrome or MPS I caused by the • Corneal clouding
deficiency of the enzyme L-iduronidase >>Hunter syndrome or MPS II caused by
>>Heparan sulphate & Dermatan Sulphate the deficiency of the enzyme L-iduronate
sulphatase
>>Accumulation in the ECM of
>>Heparan sulphate & Dermatan Sulphate
• Connective tissues – coarse facial fea-
tures, short stature >>Milder
• Organs – HSM, inguinal hernia, cardio- • No Corneal clouding
megaly • Deafness
• Neurons – Intellectual disability, retinal
>>X linked recessive
degeneration
7. Glucose and fructose are examples 10. The number of stereoisomers present
of? in a carbohydrate with n number of asymmet-
ric carbon atoms is :
A. Optical isomerism
B. Functional isomerism A. 2*n
C. Steroisomerism B. 2n
D. Epimerism C. n2
D. n3
8. Ribose and arabinose are examples 11. The various types of stereoisomerism
of? include all except,
A. Optical isomerism A. Epimerism

B. Functional isomerism B. Anomerism
C. Stereoisomerism C. Enantiomerism
Answers: 7.B; 8.D; 9.A; 4.A&D; 10.B; 11.D
D. Epimerism D. Optical isomerism
9. The most common form of glucose is:

A. β D glucopyranose
B. α D glucopyranose
C. α D glucofuranose
D. β D glucofuranose
33
Carbohydrate
Metabolism
Introduction to metabolism
• There are four steps in which substrate
• Metabolism is a process by which we as- level phosphorylation occurs
similate the food we intake. It includes
• Catabolism >Phosphoglycerate kinase
• Anabolism >Pyruvate kinase
>Succinyl thiokinase
• Catabolism is a process by which we
break down complex biomolecules into sim- >Creatine Kinase
pler substances, by breaking covalent link- • Oxidative Phosphorylation
ages. Breakage of covalent linkages causes
• The energy that is liberated is trapped as
energy to be liberated. Depending upon
NADH/ FADH2, then NADH and FADH2
how we trap the energy that is liberated,
catabolism is classifies into two types are oxidized, the liberated electrons are
transported through Electron Transport
• Substrate level phosphorylation
Chain, which in a complex way liberates
• Oxidative phosphorylation energy. This energy is used for the phos-
• Substrate level phosphorylation: phorylation of ADP to form ATP
• At the substrate level itself the energy • Examples include all dehydrogenase
that is liberated is trapped as ATP in Sub- steps which are coupled through ETC to
strate level phosphorylation generate ATP
Low Energy
• Glucose enters into glycolysis. Every Every conversion of pyruvate to acetyl
glucose is split into 2 pyruvate and this CoA provides 1 NADH, equivalent to 2.5
provides 7 ATPs ATPs
• Further fate of pyruvate is dependent on • Every Acetyl CoA enters into TCA Cycle,
whether it is aerobic or anaerobic and is converted to 2 Carbon dioxide.
• In anaerobic conditions, Lactate dehy- Every TCA cycle provides 10 ATPs
drogenase converts pyruvate to lactate. • Totally, Glucose on complete oxidation
This provides 2 ATPs provides 32 ATPs
• In aerobic conditions, Pyruvate dehydro-
genase converts pyruvate to acetyl CoA.
35
GLUCOSE
In the
Glycolysis 2 ATPS absence of
Oxygen
32 ATPS
2 PYRUVATE 2 LACTATE
2 ATPS
7 ATPS
2 ACETYL COA
In the
presence of 2 X 2.5 ATPS
Oxygen
TCA
CYCLE
High Energy
• Glucose enters into glyc- (In high energy status, liver

olysis, forms pyruvate and and muscle store glucose
pyruvate is converted to as glycogen)
acetyl CoA • Some glucose 6 phos-
• In the presence of high phate enter into HMP
energy, acetyl CoA does shunt. HMP shunt acts as
not enter into TCA cycle, a source of NADPH and
instead, it is used as a Ribose 5 Phosphate
building block of fatty acid • NADPH is used for reduc-
and cholesterol tive biosynthesis of lipid
• Fattyacids are stored as and for regenerating GSH,
triacyl glycerol and choles- which is an important
terol is stored as cholester- antioxidant mechanism of
ol ester. RBCs
• Hence, Carbohydrate • Ribose 5 phosphate is
rich diet is lipogenic used for nucleotide syn-
• In high energy status, as thesis
high energy inhibits glyco- • G6PD deficiency caus-
lysis, Glucose 6 phosphate es hemolytic anemia
becomes glucose 1 phos- following exposure to
phate, which then enters oxidative stress
into glycogen synthesis.
36 >>>
Difference between NADH & NADPH
S.No PROPERTY NADH NADPH
S.No PROPERTY NADH NADPH
1 Uses Enters ETC and Can not enter into ETC

acts as a source of 1. Acts as a coenzyme for
ATP reductive biosynthesis of lipids
2. Regeneration of GSH in RBCs
3. Ribonucleotide reductase
4. Cytochrome P450 Enzymes
5 Sources Glycolysis - 1. HMP shunt

D3PDH, TCA Cycle 2. Cytoplasmic Isocitrate
dehydrogenases, Dehydrogenase
Fatty acid oxidation 3. Malic enzyme
37
Integration of metabolism:
High Energy
NADPH
GLUCOSE
HMP SHUNT GLYCOGEN
R5P USED FOR 2 PYRUVATE

NUCLEOTIDE
SYNTHESIS
2 ACETYL COA
FATTYACID CHOLESTEROL
CARBOHYDRATE
DIET IS
LIPOGENIC!!!
CHOLESTEROL
TRIACYLGLYCEROL ESTER
Significance of glycolysis -
Preferred Fuel
fibres, renal medulla use only glucose as
• Glycolysis is the only pathway that can
their fuel
synthesis ATP anaerobically
• Aerobic cells including red muscle fi-
• Choice of fuel depends upon whether a
bres and cardiac muscle fibres use fatty
cell is aerobic or anaerobic
acid as their preferred fuel.
• Anaerobic cells including RBCs, retinal
cells and corneal cells, white muscle
• Exception is neurons, which use glucose
as their fuel, though it is aerobic.
38 >>>
Glycolysis
Definition of glycolysis
Glycolysis is a process by which a six carbon compound

glucose is split to two products. The products differ de-
pening upon whether the process occurs aerobically or
anaerobically.
• Products of aerobic glycolysis : Two molecules of Pyru-

vate and 7 ATP
• Products of anaerobic glycolysis : Two molecules of
Lactate and 2 ATP
Significance of glycolysis
1
Of the three fuels namely glu- Glycolysis is the only metabolic pathway in human metab-
cose, fatty acids and aminoacids,
olism that can produce ATP anaerobically. Hence, Glucose
glucose can be utilised both aer-
obically and anaerobically. Fatty is the only fuel that can be used anaerobically1. This is why
acids and aminoacids are utilised glycolysis is indispensible in the following cells
only aerobically.
39
2
Sodium potassium ATPase
• RBC: Mature RBCs lack mitochondria. Hence, they are
pumps out 3 sodium ions to ECF,
where the sodium concentration
dependent on a pathway to generate ATP in the ab-
is high and it takes in potassium sence of oxygen. The ATP thus produced is used for
into the ICF, where the potassium the functioning of Sodium Potassium ATPase2
concentration is high. As both the • White muscle fibres: White muscle fibres lack my-
ionic movements happen against
oglobin, the storage form of oxygen. Hence they are
concentration gradient, the pump
needs ATP. In short, the pump dependent on an anaerobic pathway to generate ATP.
maintains low sodium concentra- • Renal medulla: To maintain the medullary hyperosmo-
tion within the cell, to avoid os- lality, renal medulla receives a sparse blood supply and
motic movement of water into the hence they have relatively less access to oxygen and
cell. Hence the pump maintains
hence utilise glucose anaerobically
membrane integrity of the cell.
• Neurons: Though neurons are aerobic cells, as free
fatty acids are conjugated with albumin, they are una-
ble to cross the blood brain barrier and hence neurons
have evolved to utilise glucose preferably. The differ-
ence between neurons utilising glucose and RBC, White

muscle fibres utilising glucose is that neurons utilise
glucose aerobically and RBCs and white muscle fibres
utilise glucose anaerobically.
Steps of glycolysis
Kindly utilise the illustration of steps of glycolysis provided in

3
Glucose gets into a cell through
GLUT transporters. These trans-
the page 58, to understand the following content.
porters are of facilitated diffusion
type, which means, they can
move glucose from a region of
higher concentration to a region
Aerobic Glycolysis
of lower concentration. Initially
when the glucose concentration
is high outside the cell, GLUT
transporters move glucose into Investment Phase
the cell. In due course, if you don’t
convert glucose into any other STEP 1: As soon as glucose gets into any cell, irrespective
form, the same transporter would
of the final fate of glucose, the first enzyme to act on it is
efflux glucose back to ECF. Hence,
hexokinase or gluokinase invests hexokinase or glucokinase. Hexokinase or glucokinase con-
one ATP and convert glucose into verts 1 molecule of glucose to glucose 6 phosphate, using
glucose 6 phosphate as soon as one ATP. The purpose of this step is to trap glucose inside
glucose enters into a cell, as GLUT the cell3. This is one of the three irreversible steps of glyco-
transporters can never transport
lysis. This is one of the two steps in glycolysis, where 1 ATP
Glucose 6 phosphate .
40 >>>
KEY: HK/GK: Hexokinase/ Glucokinase, PHI: Phosphohexose isomerise,

PGK1: Phosphofructokinase1, G3PDH: Glyceraldehyde 3 phosphate
dehydrogenase, PGK: Phosphoglycerate Kinase, PGM: Phosphoglycer-
ate Mutase, PK: Pyruvate Kinase, LDH: Lactate Dehydrogenase
41
Km or Michaelis constant of an enzyme
• Km or michaelis constant is defined as the substrate

concentration at half maximal velocity.
For an understand- • Suppose km of an enzyme is 10mM, it means when
ing of differences the substrate concentration is 10mM, the enzyme can
between hexoki- achieve half maximal velocity. If km of another en-
nase and glucoki- zyme is 5mM, it means even when the substrate con-
nase, we need to centration is as low as 5mM, the enzyme can achieve

have an under- the same half maximal velocity
• If Km is low, it means the enzyme needs a lesser
standing of km of
substrate concentration to reach half maximal ve-
an enzyme.
locity. This means the affinity of the enzyme for the
substrate is high. So, Km and affinity are inversely
proportional.
Differences between hexokinase and

glucokinase:
• Hexokinase is an enzyme present in all cells whereas

glucokinase is present in liver and pancreatic beta cells.
• As liver is the first organ to meet dietary glucose , the
enzyme glucokinase present in liver exhibits lower affin-
ity or higher km (20mM). Had liver been provided with
an enzyme with higher affinity or lower km, liver would
have utilised every molecule of dietary glucose and the
periphery would have suffered from hypoglycaemia.
• In pancreatic beta cells, as soon as glucose4 enters into
the cell, it enters into glycolysis and citric acid cycle. ATP
is generated. ATP closes ATP sensitive potassium chan-
nels. This causes depolarisation and action potential.
Action potential opens voltage gated calcium channels.
4
Glucose in the diet gets absorbed
into portal vein and reaches liver first This calcium entry causes insulin release.
42 >>>
STEPS INVOLVED IN INSULIN RELEASE
KEY: K : ATP sensitive potassium channels, Ca: voltage gat-

ed calcium channels
• This is primarily why glucokinase in pancreatic β cells
exhibits lower affinity and higher km for its substrate glu-
cose. If only glucokinase in pancreatic β cells is provided
5
Induction of an enzyme with higher affinity, the sequence of events which lead to
is regulation of enzyme at insulin release would have happened even at low glu-
transcription level. If a reg-
ulator increases the rate of cose and insulin would have aggravated hypoglycaemia
transcription of the gene of • Hexokinase is inhibited by its product glucose 6 phos-
an enzyme, increasing the phate, whereas glucokinase is not inhibited by glucose 6
number of enzymes pro- phosphate
duced, thereby collectively
• Glucokinase is inducible5 by insulin but hexokinase is not
increasing an enzyme activi-
ty, it is called as induction induced by insulin.
Differences between hexokinase and

glucokinase
43
S.No PROPERTY HEXOKINASE GLUCOKINASE
1 Location All cells Liver, Pancreatic β cells
2 Km and Higher affinity and low Lower affinity and high

affinity km Km = 0.2mM Km Km = 10mM
3 Inhibition by Inhibited by glucose 6 Not inhibited by glucose

the product phosphate 6 phosphate
glucose 6
phosphate
4 Induction by Not induced by insulin Induced by insulin

Insulin
STEP 2: Glucose 6 phosphate is converted to fructose 6

phosphate by phosphohexose isomerase. It is a reversible
step.
STEP 3: Fructose 6 phosphate is converted to fructose 1,6

bisphosphate by phosphofructokinase I. This step utilises
one ATP. Hence this is a step where ATP is invested in glyc-
olysis. It is an irreversible step of glycolysis. This step is
considered as the rate limiting step of glycolysis.
With this, investment phase of glycolysis is over. In this

phase, for every molecule of glucose, 2 ATPs are utilised.
Harvesting phase
STEP 4: Fructose 1,6 bisphosphate is cleaved by Aldolase

A to form two trioses -Glyceraldehyde 3 phosphate and
Dihydroxyacetone phosphate.
STEP 5: Glyceraldehyde 3 phosphate (G3P) and Dihy-
droxyacetone Phosphate (DHAP) are trioses and they are
isomers and they are interconvertible by Phosphotriose
isomerise.
STEP 6: G3P is converted to 1,3 bisphosphoglycerate by
Glyceraldehyde 3 phosphate dehydrogenase (G3PDH). As
44 >>>
G3P is converted to subsequent products, the equilibrium

between G3P and DHAP in step 5, gets shifted towards
forming G3P. Hence every DHAP formed in step 4 is di-
verted towards forming G3P. Hence, two molecules of G3P
participate in this chemical reaction, forming two mole-
cules of 1,3 bisphosphoglycerate.
In this step, G3PDH, as the name suggests removes hydro-
gen from the substrate - Glyceraldehyde 3 phosphate and
the hydrogen is provided to NAD to form NADH. Hence,
from two molecules of G3P, 2 NADH are formed. When
electrons from these 2 NADH enter into electron transport
chain, 5 ATPs6 are formed. Hence, this is a step in which
oxidative phosphorylation occurs in glycolysis.
6
1 NADH provides 2.5 ATPs STEP 7: Two molecules of 1,3 Bisphosphoglycerate are
through oxidative phsophorylation converted to two molecules of 3 phosphoglycerate by
phosphoglycerate kinase. 1,3 bisphosphoglycerate has a
high energy linkage and when the linkage is broken, the
energy liberated is high enough to be coupled to ATP
synthesis. This step is thus coupled to the generation of
2 molecules of ATP. Hence, this is a substrate level
phosphorylation step.
>> This step is bypassed in RBCs through Rapaport
Leubering shunt, which will be discussed in subsequent
sections.
STEP 8: Two molecules of 3-Phosphoglycerate are con-
verted to two molecules of 2-phosphoglycerate by a
positional isomerism step. Mutase is an enzyme which
catalyses positional isomerism steps. Hence, the enzyme
involved in this step is phosphoglycerate mutase
STEP 9: Two molecules of 2-phosphoglycerate lose a
molecule of water each, in the presence of enolase to form
two molecules of phosphoenol pyruvate
STEP 10: Two molecules of Phosphoenol pyruvate are
converted to two molecules of pyruvate by pyruvate
kinase. This is an irreversible step. Phosphoenol pyru-
vate has a high energy linkage. Hence, cleavage of the link-
age is coupled to ATP production. This step is an example
of substrate level phosphorylation and it generates two
molecules of ATP.
Thus, as a result of harvesting phase, 9 ATP are generated..
The net ATP produced in aerobic glycolysis is 9-2 =7 ATPs
45
Anaerobic glycolysis
The one step of aerobic in aerobic glycolysis. Hence is regenerated from NADH
glycolysis which is affected in anaerobic glycolysis, we by Lactate dehydrogenase.
in anaerobic conditions is get only 2 ATPs (7 in aerobic Lactate dehydrogenase
Glyceraldehyde 3 phos- glycolysis -5) converts 2 molecules of
phate dehydrogenase Furthermore, for want of Pyruvate to 2 molecules of
step. In the absence of oxygen as NADH is not lactate and for every con-
oxygen, electrons from the oxidised back to NAD, NAD version 2 NADH get regener-
two molecules of NADH gen- deficiency is expected in ated as NAD, allowing glyco-
erated in this step will not anaerobic glycolysis. With- lysis to continue even in the
be able to get into electron out NAD, glyceraldehyde 3 absence of oxygen.
transport chain. Hence, in phosphate dehydrogenase Hence, products of anaer-

anaerobic glycolysis, we becomes inactive and glyc- obic glycolysis are 2 mole-
would be missing the 5 mol- olysis would stop. To avoid cules of lactate and 2 ATP.
ecules of ATP, which we get this, in anaerobic cells, NAD
46 >>>
Rapaport Leubering cycle

It is otherwise called as 2,3 Bisphosphoglycerate shunt.
Significance:
This shunt is operative in RBCs and it acts as a source of 2,3
Bisphosphoglycerate (2,3 BPG). 2,3 BPG reduces the affinity
of Hemoglobin for oxygen and helps in unloading oxygen
to tissues. Hence, 2,3 BPG must be generated in RBCs in
hypoxia and anemia.
Pathway:
GLYCOLYSIS RAPAPORT LEUBERING SHUNT
Glyceraldehyde 3 phosphate dehydrogenase acting on 2

molecules of glyceraldehyde 3 phosphate (G3P). 1,3 BPG
is then acted upon by phosphoglycerate kinase to form 3
phosphoglycerate (3-PG). It is a substrate level phosphoryl-
ation step. Hence in this step, 2 ATPs get generated
2,3 BPG shunt:
In conditions of hypoxia and anemia, there is a deviation of
glycolysis in RBCs to generate 2,3BPG. To convert 1,3 BPG
to 2,3 BPG, bisphosphoglycerate mutase acts. 2,3BPG then
reduces the affinity of hemoglobin and helps in unloading
oxygen to tissues. 2,3 BPG is then acted upon by Bispho-
glycerate phosphatase and is converted to 3 phosphoglyc-
erate.
47
Hence, this pathway begins with an intermediate of glyco-

lysis (1,3 BPG), generates 2,3 BPG for optimal functioning
of RBCs and gets back to the next intermediate of glycoly-
sis – 3 phosphoglycerate. Therefore it is called as a shunt
– Rapaport Leubering shunt or 2,3 BPG shunt.
Advantage:
This shunt acts as a source of 2,3 BPG
Disadvantage:
As RBCs do not have mitochondria, glucose is utilised
through anaerobic glycolysis. Hence from a molecule of
glucose, 2 ATPs get generated normally. When RBCs go
through Rapaport Leubering Shunt, because the substrate
level phosphorylation step catalysed by phosphoglycerate
kinase step is by passed, nil ATP production is observed in
RBCs.
Regulation of glycolysis
Glycolysis is regulated at the three irre- Hexokinase or Glucokinase :
versible steps of glycolysis: Hexokinase is inhibited by the product glu-
1. Hexokinase or Glucokinase step cose 6 phosphate. Hence,
2. Phosphofructokinase I step
3. Pyruvate Kinase step
Summary
• Metabolism has two components – catabolism and
anabolism
• Catabolism is of two types – Substrate Level phospho-
rylation and oxidative phosphorylation
• Steps catalysed by Phosphoglycerate kinase, pyruvate
kinase enzymes of glycolysis, succinyl thiokinase en-
zyme of citric acid cycle and creatine kinase of muscle
are the four examples of substrate level phosphorylation
• All other catabolic steps are of oxidative phosphoryla-
tion type.
• Gluconeogenesis is an anabolic pathway
• Citric acid cycle is an amphibolic pathway
48 >>>
Hemolytic Anemia - Metabolic Causes
>> G6PD deficiency – Hemolytic anemia >> Glycolytic enzyme defect – Hemolytic
following exposure to oxidative stress anemia + Exercise intolerance
• PFK1 Defect – Low 2,3 BPG
• Pyruvate Kinase Defect – High 2,3 BPG
MCQS
1. Primaquine can precipitate hemolyt- 3. Iodoacetate inhibits which of the follow-
ic anemia in individuals with an enzyme ing enzymes?
deficiency. The enzyme is related to which
pathway? A. Phosphoglycerate kinase
B. Pyruvate kinase
A. Gluconeogenesis C. Enolase
B. Glycolysis D. Glyceraldehyde 3 phosphate dehydro-
C. HMP shunt genase
4. True about Rapaport Leubering shunt is?

D. Glycogen metabolism
2. True about Glucokinase is?

A. 2 ATP produced
A. Present in kidney B. Uses phosphoglycerate kinase enzyme
B. Low Km C. Acts as a source of NADPH
C. Induced by Insulin D. Stimulated in anemia
D. Inhibited by Glucose 6 phosphate
49
Integrated case based

5. 12. A neonate presented with hemo-
lytic anemia. Peripheral smear revealed
that it’s a case of non spherocytic hemo-
lytic anemia. Pyruvate kinase activity was
remarkably low (0.675 U/g Hb). Which of
the following explains hemolytic anemia in
this condition?
A. Low 2,3 BPG production

B. High 2,3 BPG production
C. High ATP production

D. Failure of Sodium Potassium ATPase pump
Image based MCQS

6. Name a chemical which inhibits the
enzyme which is shown as red square in
the given picture.
A. Iodoacetate
B. Arsenate
C. Arsenite
D. Fluride
Answers: 1.C; 2.C; 3.D; 4.D; 5.D; 6.D
50 >>>
Fates of pyruvate
Introduction
• Pyruvate is a central metabolite and it can • An understanding of all the fates of pyru-
have various fates, the choice of the fate vate is easier if we understand what hap-
depends upon the energy status of the cell. pens in starvation
Changes in starvation
• Starvation causes deficiency of all 3 fuels – • Liver Glycogenolysis begins 2 to 2 and a
glucose, fatty acid and aminoacid. half hours after the previous meal and main-
• Glucose deficiency is more worrisome, as tain plasma glucose till 12 to 18 hours of
principal cells of our body namely neurons starvation
and RBCs are dependent on glucose • Gluconeogenesis starts 6 hours after the
• Liver Glycogenolysis and Gluconeogenesis previous meal and needs a substrate and
maintain plasma glucose energy, both of which are supplied by periph-
eral lipolysis
Starvation and peripheral lipolysis

• Low glucose and Low insulin in hypogly- • Both glycerol and free fatty acid through
cemia stimulate hormone sensitive lipase in circulation reach the liver
adipose tissue • Glycerol is used as a substrate for gluco-
• Hormone sensitive lipase cleaves adipose neogenesis
tissue triacyl glycerol to form glycerol and • Fatty acids are oxidized to provide the
free fatty acid energy for gluconeogenesis
Fatty acid oxidation defects cause

hypoglycemia
51
Fates of pyruvete
Fates of pyruvate depend upon the energy status of the cell.
Low energy status: High Energy Status:

When the energy status of the cell is low, When the energy status of the cell is high,
the fate of pyruvate is dependent on wheth- the fate of pyruvate is dependent on wheth-
er the cell is aerobic or anaerobic. When er the person is well fed or if the person is
the cell is anaerobic, for anaerobic glycol- in starvation. If the person is in starvation
ysis to continue, Pyruvate is converted to and if the cells are in high energy status,
Lactate by Lactate Dehydrogenase. When it means the cell is striving hard for gluco-
the cell is aerobic, pyruvate is converted neogenesis. Hence pyruvate is converted
to acetyl CoA by Pyruvate dehydrogenase to Oxaloacetate by Pyruvate carboxylase.

complex, so that acetyl CoA can enter into When the person is well fed and if the ener-
citric acid cycle to provide more energy gy status is high, pyruvate should be used
for anabolism. Hence alanine transaminase
or serum Glutamate pyruvate transaminase
converts pyruvate to alanine.
LOW ENERGY HIGH ENERGY
AEROBIC ANAEROBIC FED STATE STARVATION

Carboxylase
ALT/SGPT
LDH
PDH
Py
ACETYL CoA LACTATE ALANINE OXALOACETATE
52 >>>
Pyruvate
dehydrogenase
Facts about PDH complex
product. Hence Acetyl CoA, NADH & CO2
• The complex is present in mitochondria
are products
• It catalyses an irreversible step
• Pyruvate, NAD & CoA are substrates
• It catalyses an Oxidative decarboxylation
reaction
• It has three subunits – PDH, Dihydrolipoyl
Transacetylase, Dihydrolipoyl dehydroge-
• When it oxidises, NAD is converted to nase
NADH, when it decarboxylates, Carbon di-
oxide is release and Acetyl CoA is the major
• It uses five Coenzymes – TPP, Lipoamide,
CoA, FAD, NAD
• It is inhibited by Arsenite
Regulation of PDH complex
ALLOSTERIC REGULATION COVALENT MODIFICATION INDUCTION OR REPRES-

• Inhibited by acetyl CoA, • Stimulated by Dephos- SION
NADH and ATP phorylation • Induced by Insulin in
adipose tissue
Glucose sparing effect

>> The body tries to act like a mother with >> In our body, the adamant child role is
two children. One child is adamant and the played by neurons and RBCs. They are
child wants only chips as his or her snack. adamant that they can use only glucose as
The other child is cool and is happy taking their fuel. The cool child role is played by
an apple or chips for his or her snack. In cardiac muscle fibres and red muscle fibres.
this case, the mother spares the chips for They are cool because they can use either
the adamant child to avoid temper tantrum glucose or free fatty acid as their fuel. So,
53
the body spares glucose from being used >> Citrate and ATP inhibit Phosphofructoki-
by cells which can utilize fatty acids. nase I – the rate limiting enzyme of Glycol-
>> Glucose sparing effect is inhibition of ysis
glucose oxidation by fatty acid oxidation >> Acetyl CoA, NADH ad ATP stimulate
products. It is shown by three regulation Pyruvate Carboxylase – the rate limiting
>> Acetyl CoA, NADH and ATP being prod- enzyme of gluconeogenesie
ucts of fatty acid oxidation, inhibit glucose
oxidation by inhibiting PDH complex
MOTHER
ADAMANT COOL
CHIPS APPLE CHIPS

• Inhibition of glucose
oxidation by fatty
acid oxidation
• Acetyl CoA, NADH
and ATP inhibt PDH MOTHER
complex
• Citrate and ATP
inhibt PFK1 NEURONS & RBCs CARDIAC MUSCLE FIBRES
• Acetyl CoA,
NADH and ATP
stimulate Pyruvate GLUCOSE FATTY ACID GLUCOSE
carboxylase
Pasteur effect
with aerobic and anaerobic terms, it is Pas-
• Pasteur effect is a modification of glucose
teur effect
sparing effect. Glucose can be oxidised an-
aerobically but fatty acids can be oxidised • Pasteur effect is inhibition of anaer-
aerobically obic oxidation by aerobic oxidation
• Hence instead of phrasing it as inhibition products
of glucose oxidation by fatty acid oxidation • Citrate and ATP inhibit Phosphofructoki-
(glucose sparing effect), if it is rephrased nase I – the rate limiting enzyme of Glycol-
ysis
54 >>>
BODY
CARDIAC MUSCLE
NEURONS & RBCs
FIBRES
GLUCOSE FATTY ACID GLUCOSE
INHIBITION OF ANAEROBIC OXIDATION BY AEROBIC OXIDATION
Citrate and ATP inhibit PFK1
Warburg effect
• Neoplastic cells’ metabolic machinery is reprogrammed
to utilise aerobic glycolysis
• Quicker ATP production

• Intermediates of glycolysis form building blocks essential
for growth and multiplication of neoplastic cells
• NADPH generated protects neoplastic cells from antican-

cer drugs
Ketogenic Diet
• High fatty diet with low carbs

• Low carbohydrates causes low glucose and hence neo-
plastic cells do not get a good access to glucose
• Also fatty acids on oxidation form citrate and ATP, which

inhibit PFK1. Hence neoplastic cells can not thrive
55
MCQS
1. All the following are fates of pyruvate 2. All the following are coenzymes of
except? pyruvate Dehydrogenase complex except?
A. Acetyl CoA A. Thiamine

B. Lactate B. Riboflavin
C. Alanine C. Niacin
D. Valine D. Biotin
Image based MCQS

3. Identify E1 and E2 Pyruvate Dehydrogenase

Complex Enzymes
A. Pyruvate decarboxylase, Dehydroge-
nase
B. Pyruvate decarboxylase, Transacetylase
C. Transacetylase, Dehydrogenase
D. Dehydrogenase, decarboxylase
4. A 34 year old male is diagnosed with

papillary carcinoma of thyroid. He is treated
with thyroidectomy. His wife learns online
that ketogenic diet can avoid neoplasia.
Which of the following regulatory effects
best explain the antineoplastic effect of
ketogenic diet?
A. Acetyl CoA stimulates Pyruvate carbox-

Answers: 1.D; 2.D; 3.B; 4.C.
ylaseb.
B. Malonyl CoA inhibits Carnitine Acyl
Transferase Ic.
C. Citrate inhibits PFK1d.
D. Acyl CoA stimulates Carnitine Acyl Trans-
ferase I
56 >>>
TCA cycle
Few facts about TCA cycle
• It is a mitochondrial process • Four oxidative phosphorylation steps –
• It is strictly aerobic Isocitrate dehydrogenase, Alpha Ketogluta-
• As NAD & FAD can not be regenerated rate dehydrogenase, Succinate Dehydroge-
without Oxygen nase, Malate dehydrogenase steps
• Acetyl CoA and oxaloacetate enter TCA cy- • One substrate level phosphorylation step
cle and Oxaloacetate is regenerated, closing – Succinyl thiokinase step
the cycle. The two carbon atoms of acetyl • Two Oxidative decarboxylation steps -
CoA come out as 2 Carbon dioxide. Hence ICDH, AKGDH
TCA cycle is considered as a complete oxida- • One FAD linked dehydrogenase – Succi-
tion process, where all fuels get oxidised. nate Dehydrogenase
• Every TCA cycle provides 10 ATPs
Steps of citric acid cycle
57
Oxidative decarboxylation steps

• Pyruvate dehydrogenase • Alpha Ketoglutarate dehy- • Branched Chain Ketoacid
• Isocitrate dehydrogenase drogenase Dehydrogenase of Branched
• 6 Phosphogluconate de- chain amino acid metabolism
hydrogenase of HMP shunt
FAD linked dehydrogenase

• Succinate Dehydrogenase • Mitochondrial Glycerol 3
• Acyl CoA dehydrogenase phosphate dehydrogenase
Regulation of TCA cycle

• All steps are stimulated by low energy • The direction depends upon the energy
status indicators – NAD, FAD, ADP status of the cell
• All dehyrogenases are stimulated by calci- • When the energy status of the cell is low,
um the pathway occurs in clockwise direction,
• The chief rate limiting enzyme is Isocitrate generating 10 ATPs per cycle. When the
dehydrogenase energy status of the cell is high, all enzymes
of the pathway is inhibited. The intermedi-
• TCA Cycle is an amphibolic pathway. This ates accumulate because of anaplerotic
cycle can occur in catabolic direction or in
reactions and the intermediates are used
an anabolic direction
for anabolism
Anaplerotic reactions
These are filling up reactions, which fill up • Fumarate is not only formed from succi-
the intermediates. nate, it is also formed from phe and Tyr
Examples: • The most important anaplerotic reaction is
• Alphaketoglutarate is not only formed catalysed by malic enzyme which converts
from isocitrate, but also from glu, gln, His, malate to pyruvate. Because from Pyru-
pro and Arg vate, we can form both acetyl CoA and
• Succinyl CoA is not only formed from Oxaloacetate
alpha ketoglutarate, but also from Val, Ile
and Met, odd chain fatty acids
58 >>>
Significance of malic enzyme

>> Best anaplerotic reaction >> One of the 3 sources of
of TCA Cycle NADPH
Steps of citric acid cycle
Anaplerotic reactions
Pyr
Malic
enzyme
Phe
Tyr
Val Glu
Ile Gln
Met His
Pro
Arg
59
MCQS
1. All the following are examples of 5. All the following are examples of oxida-
oxidative phosphorylation step enzymes tive phosphorylation step enzymes except
except
A. Isocitrate dehydrogenase
A. Isocitrate dehydrogenase B. Alpha ketoglutarate dehydrogenase
B. Alpha ketoglutarate dehydrogenase C. Succinyl thiokinase
C. Succinyl thiokinase D. Succinate dehydrogenase
6. The rate limiting enzyme of citric acid

D. Succinate dehydrogenase
2. The rate limiting enzyme of citric acid

cycle is
cycle is A. Isocitrate dehydrogenase

B. Alpha ketoglutarate dehydrogenase
A. Isocitrate dehydrogenase C. Succinyl thiokinase
B. Alpha ketoglutarate dehydrogenase D. Succinate dehydrogenase
7. All the following are stimulators of Isoc-

C. Succinyl thiokinase
D. Succinate dehydrogenase
3. All the following are stimulators of Isoc-

itrate dehydrogenase except
itrate dehydrogenase except A. NAD

B. FAD
A. NAD C. ATP
B. FAD D. Calcium
8. Oxidative decarboxylation reactions are

C. ATP
D. Calcium
4. Oxidative decarboxylation reactions are

Answers: 1.C; 2.A; 3.C; 4.D; 5.C; 6.A; 7.C; 8.D.
catalyzed by all, EXCEPT:
catalyzed by all, EXCEPT: A. PDH

B. Isocitrate dehydrogenase
A. PDH C. α Ketoglutarate dehydrogenase
B. Isocitrate dehydrogenase D. Malate dehydrogenase
C. α Ketoglutarate dehydrogenase
D. Malate dehydrogenase
60 >>>
Glycogen metabolism
Facts to remember
• Glycogen is stored in liver and muscle • Total Glycogen mass is higher in muscle
• Glycogen content or concentration (g/kg as the muscle mass is more.
of tissue) is highest in liver • Glycogen metabolism includes Glycogen
synthesis and Glycogenolysis
Steps involved in glycogen synthesis
• Glucose is converted to • UDP glucose is attached to

glucose 6 phosphate by glycogenin forming glycogen
hexokinase in muscle and primer by glycogen synthase
glucokinase in liver. This step • This chain grows until 11
uses one high energy phos- to 13 glucose residues get
phate attached in a straight chain
• Glucose 6 phosphate is • Branching enzyme removes
converted to glucose 1 phos- peripheral 6 glucose resi-
phate by phosphoglucomu- dues from a straight chain
tase [attached with alpha(1,4)
• Glucose 1 phosphate is linkage] and attached it to
activated to for UDP glucose another chain with alpha(1,6)
by UDP glucose pyrophos- linkage
phorylase. This step uses 2 • This chain grows until there
high energy phosphates. are 11 to 13 glucose residues.
• Hence 3 high energy phos- Then branching enzyme re-
phates are used for adding peats the action to generate
one molecule to growing alpha(1,6) linkage
glycogen
61
Glycogen synthesis
GLYCOGEN
GLYCOGENIN BRANCHING ENZYME
GLYCOGEN SYNTHASE
UDP- Glu
UDP GLU
UTP → UMP
PYROPHOSPHORYLASE
GLUCOSE 1
PHOSPHATE
PHOSPHOGLUCO
MUTASE
GLUCOSE 6
PHOSPHATE
ATP / GLUCOKINASE
GLUCOSE
Facts to remember about

glycogen synthesis
• Liver and Muscle • Branching enzyme is α (1,4) → (1,6) glu-
• UDP glucose is the active form of glucose can transferase
• Three high energy phosphates are re- • Glycogen synthase is the rate limiting
quired for attaching every glucose enzyme
• It gets activated by dephosphorylation
Enzymes activated by
phosphorylation
Glucagon acts by phosphorylation. Hence enzymes
which increase plasma glucose get activated by phos-
phorylation. Glycogenolysis and Gluconeogenesis are
the two pathways get stimulated by phosphorylation.
62 >>>
ENZYMES OF GLYCOGENOLYSIS: ENZYMES OF GLUCONEOGENESIS:

• Glycogen phosphorylase • Pyruvate carboxylase
• Debranching enzyme • PEPCK
• Fructose 1,6 bisphosphatase
• Glucose 6 phosphatase
Hormone sensitive lipaseUr simostiatiur
aribusdant, occum et volo volectempos
Glycogenolysis
Steps of glycogenolysis
• Glycogen phosphorylase • So in liver, free glucose is
adds inorganic phosphate formed, which reaches cir-
to glycogen with n num- culation increasing plasma
ber of glucose residues to glucose
form glucose 1 phosphate • Muscle glycogenolysis
and glycogen with (n-1) product is glucose 1
glucose residues phosphate, which enters
• So immediate product of into glycolysis and is used
glycogenolysis is glucose for muscle purposes.
1 phosphate • Glycogen phosphorylase
• Glucose 1 phosphate is can cleave alpha (1,4)
converted to glucose 6 linkage and hence it re-
phosphate by phospho- moves peripheral glucose
glucomutase residues until there are 4
• Until this step, the steps residues in a chain
involved in liver glycogen- • Debranching enzyme
olysis and muscle glycog- cleaves 3 glucose resi-
enolysis are the same dues and transfers the
• As liver glycogenolysis is a trisaccharide to another
gluconeogenic organ, it is chain. This exposes alpha
equipped with glucose 6 (1,6) linkage
phosphatase • Debranching enzyme
breaks alpha (1,6) linkage
63
Glycogenolysis
Debranching enzyme
64 >>>

glycogenolysis
• Liver glycogenolysis in- provides one ATP more
creases plasma glucose than that is produced by
but muscle glycogen- muscle glycogenolysis
olysis can not increase • It is activated by phos-
plasma glucose phorylation
• Glycogen phosphorylase • Debranching enzyme is
requires inorganic phos- α (1,4) → α (1,4) glucan
phate to be active transferase
• Glucose produced by
glycogenolysis in muscle
Diferences between liver and

muscle phosphorylase
LIVER GLYCOGEN MUSCLE GLYCOGEN

S.No PROPERTY
PHOSPHORYLASE PHOSPHORYLASE
65
LIVER GLYCOGEN MUSCLE GLYCOGEN

S.No PROPERTY
PHOSPHORYLASE PHOSPHORYLASE
1 Products Glucose 6 phosphate, Glucose 6 phosphate & ATP

Glucose, ATP
2 Inhibitor Inhibited by all products Inhibited by all products

and hence not inhibited
by glucose
3 Stimulated by Glucagon and Epinephrine Epinephrine and calcium
Glycogen phosphorylase-
Regulation
• Enzyme exists in two forms - • Glycogen phosphorylase b • b is converted to a by cal-

a and b is converted to a by phos- cium and cAMP dependent
• a is active (phosphorylated phorylase kinase kinase
form) • Phosphorylase kinase exists
• b is inactive (dephosphoryl- in two forms – a (active) and
ated form) b (inactive)
Number of ATPs Number of ATPs from

S.No Pathway from 1 molecule 1 molecule of glucose
of free glucose from glycogenolysis
1 Aerobic glycolysis
2 Anaerobic glycolysis
3 Complete oxidation
Number of ATPs Number of ATPs from

S.N o
Pathway from 1 molecule 1 molecule of glucose
of free glucose from glycogenolysis
1 Aerobic glycolysis 7 8
2 Anaerobic glycolysis 2 3
3 Complete oxidation 32 33
66 >>>
MCQS
1. The organ with maximum glycogen 5. The rate limiting enzyme of glycogenoly-
concentration is: sis is
A. Liver A. Glycogen synthase

B. Skeletal muscle B. Glycogen phosphorylase
C. Cardiac muscle C. Glucose 6 phosphatase
D. Neuron D. Debranching enzyme
6. The immediate product of glycogen

2. The active form of glucose is: phosphorylase is
A. β D Glucopyranose A. Glucose
B. α D Glucopyranose B. Glucose 1 phosphate
C. UDP glucose C. Glucose 6 phosphate
D. Glucose 6 phosphate D. Dextran
3. The branching enzyme in glycogen syn- 7. ASSERTION: Muscle glycogenolysis can

thesis is: not increase blood glucose
REASON: Muscle lacks glucose 6 phosphatase
A. α(1,4) → α(1,4) glucan transferase
B. α(1,4) → α(1,6) glucan transferase A. Both Assertion and Reason are true, and Rea-
C. α(1,6) → α(1,4) glucan transferase son is the correct explanation for Assertion
D. α(1,4) → β(1,4) glucan transferase B. Both Assertion and Reason are true, and
4. The enzyme which gets activated by

Reason is not the correct explanation
for assertion
dephosphorylation is: C. Assertion is true, but Reason is false
D. Assertion is false, but Reason is true
8. Glucagon stimulates:
A. Glycogen synthase
B. Glycogen Phosphorylase
C. Fructose 1,6 bisphosphatase
D. Hormone sensitive lipase A. Liver glycogen phosphorylase
B. Muscle glycogen phosphorylase
C. Both
D. None
67
9. Epinephrine stimulates: 11. The number of ATP produced from

complete oxidation of glucose obtained from
A. Liver glycogen phosphorylase muscle glycogenolysis is
B. Muscle glycogen phosphorylase
C. Both A. 32
D. None B. 33
C. 7
10. Glycogen phosphorylase a is con-

D. 8
verted to b by
A. Insulin
B. calcium
C. cAMP
D. Phosphorylase kinase a
Answers: 1.A; 2.C; 3.B; 4.A; 5.B; 6.B; 7.A; 8.A; 9.C; 10.A; 11.B
68 >>>
Glycogen storage
disorders
S.No Type Name Enzyme defect
1 0
2 I
3 II
4 III
5 IV
6 V
7 VI
8 VII
69
S.No Type Name Enzyme defect
1 0 Glycogen Synthase Glycogen Synthase
2 I Von Gierke’s disease Glucose 6 phosphatse
3 II Pompe’s disease Acid maltase
4 III Cori’s disease Debranching enzyme
5 IV Anderson’s disease Branching enzyme
6 V Mc Ardle’s disease Muscle Phosphorylase
7 VI Her’s disease Liver Phosphorylase
8 VII Tarui’s disease Phosphofructokinase I

Pompe’s disease
• It is caused by the defect of Acid mal- • When cardiac muscle gets affected, it
tase or alpha glucosidase defect.It is the causes Cardiomegaly, Cardiomyopathy.
only glycogen storage disorder which is When skeletal muscle gets affected, it
also a lysosomal storage disorder presents as hypotonia, respiratory dis-
• 2% glycogen is metabolised in skeletal tress and failure to thrive. The course of
muscle and cardiac muscle by lyso- the illness is rapid and death happens
somes with the help of acid maltase or mostly close to 2 years
alpha glucosidase • Enzyme replacement therapy is available
Presentation of glycogen
storage disorders
• Glycogen storage disorders present with • Disorders which present with hypoglycemia
hypoglycemia or exercise intolerance or both • Type 0 (Glycogen Synthase defect)
• Those disorders which are caused by a de- • Type I
fect of liver glycogenolysis or gluconeogene- • Type III
sis or both present with hypoglycemia • Type VI
• Those disorders which are caused by a • Fanconi Bickel Syndrome
defect of muscle glycogenolysis or glycolysis • Disorders which present with exercise
present with exercise intolerance intolerance
70 >>>
• Type 0 (Glycogen Synthase defect) • Type 0 (Glycogen Synthase defect)

• Type III • Type III
• Type V • Disorders which present with neither hypo-
• Type VII glycemia nor exercise intolerance
• Disorders which present with both hypo- • Type IV
glycemia and exercise intolerance
Glycogen storage disorders

with hypoglycemia
S.No Type Name Features
1 0 Glycogen Synthase Hypoglycemia and

defect exercise intolerance. Zero
glycogen in liver
2 I Von Gierke’s disease Hypoglycemia which

does not respond to
counter regulatory
hormone administration
3 III Cori’s disease Hypoglycemia + Exercise

intolerance. Abnormal
glycogen with multiple
branch points in liver
4 VI Her’s disease Hypoglycemia
5 VII Fanconi Bickel Syndrome Hypoglycemia, Polyuria,

Polydypsia, Short stature
Responds
No other to counter
Her’s regulatory
features
hormone
Von Gierke’s
No response
Exercise to counter
HYPOGLYCEMIA Intolerance regulatory
Cori’sDisease hormone
Type 0
Glycosuria,
Polyuria, Short
stature FanconiBickel
71
Von Gierke’s Disease
• Type I Glycogen storage • The accumulated Glu- • Hyper triglyceridemia is

disorder. It is caused by cose 6 phosphate enters the reason for renomega-
a defect of Glucose 6 glycolysis and forms ly, Doll like faces
phosphatase pyruvate. Pyruvate is • Hypercholesterolemia is
• Glucose 6 phosphatase converted to acetyl CoA. the reason for Xanthom-
is an enzyme of glycog- Acetyl CoA is used for as
enolysis and gluconeo- fatty acid and cholesterol • Acetyl CoA condenses
genesis. As just these two synthesis. Fatty acid is to form ketone bodies,
pathways can increase stored as triacyl glycerol hence ketosis is ob-
blood glucose, it pre- and cholesterol ester. served

sents as hypoglycemia Hence the child also
• Pyruvate is converted to
which does not respond presents with hypertri-
lactate and hence lactic
to counter regulatory glyceridemia and hyper
acidosis
hormone administration cholesterolemia.
Glycogen storage disorders

with exercise intolerance
72 >>>
1 0 Glycogen Synthase Hypoglycemia + Exercise

defect intolerance
0 glycogen in liver
2 III Cori’s disease Hypoglycemia + Exercise

intolerance
3 V Mc Ardle’s disease Exercise Intolerance
4 VII Tarui’s disease Exercise Intolerance +

Hemolytic anemia
Hemolytic Tarui’s
anemia
With
Cori’s Disease
Exercise Hypoglycemia
intolerance
Type 0
No hemolytic
anemia Mc Ardle’s
MCQS
1. The glycogen storage disorder which 2. The glycogen storage disorder
is also a lysosomal storage disorder is caused by the defect of muscle phospho-
rylase is
A. Pompe’s disease
B. McArdles disease A. Pompe’s disease
C. Hers disease B. McArdles disease
D. Taruis disease C. Hers disease
D. Taruis disease
73
3. The glycogen storage disorder which 6. The glycogen storage disorder which
does not present with hypoglycemia is presents with neither hypoglycemia nor exer-
cise intolerance is
A. Cori’s disease
B. McArdles disease A. Cori’s disease
C. Hers disease B. Anderson’s disease
D. Von Gierke’s disease C. Mc Ardle’s disease
4. A 8 month old infant presents with

D. Von Gierke’s disease
hypoglycemia. On examination, hepatomegaly 7. The glycogen storage disorder which

was observed. Blood investigations revealed presents with hypoglycemia, which does not
lactic acidosis, ketosis and Xanthomas on the respond to counter regulatory hormone ad-
buttocks. What is the most probable enzyme ministration is
deficiency?
A. Cori’s disease
A. Branching enzyme B. Fanconi Bickel’s syndrome

B. Glycogen Synthase C. Mc Ardle’s disease
C. Acid Maltase D. Von Gierke’s disease
D. Glucose 6 Phosphatase
5. A person presents with intolerance to an-

aerobic exercises and he says he experience
pain just following 5 minutes of anaerobic
exercises. His plasma glucose is normal. LDH
is normal. The most probable cause is
A. Cori’s disease
B. McArdles disease
C. Hers disease
D. Tarui’s disease
Answers: 1.A; 2.B; 3.B; 4.D; 5.B; 6.B; 7.D.
74 >>>
Gluconeogenesis

Gluconeogenesis
>> Gluconeogenesis is a pathway by which • PEPCK – uses two GTP
we synthesise glucose from non carbohy- • Phosphoglycerate kinase – uses 2 high
drate precursors. It is reversal of glycolysis. energy phosphates
In glycolysis, we convert glucose into 2 • Glyceraldehyde 3 phosphate dehydroge-
pyruvate. In gluconeogenesis, 2 pyruvate nase uses 2 NADH (5 ATPS)
molecules are converted into 1 glucose
>> Substances which on catabolism can
>> It happens in Liver and kidney give rise to either a glycolytic or a TCA cycle
>> Enzymes involved: As glycolysis is a intermediate other than acetyl CoA can act
reversal of glycolysis, enzymes which reverse as gluconeogenic substrates.
the following irreversible steps of glycolysis >> Glycerol, lactate and Alanine are the 3
are enzymes of gluconeogenesis principal gluconeogenic substrates
• Hexokinase or Glucokinase >> Acetyl CoA is never gluconeogenic as
• Phosphofructokinase I PDH complex is irreversible
• Pyruvate Kinase >> Hence even chain fatty acids are never
>> Enzymes involved in gluconeogenesis: gluconeogenic
• Pyruvate Carboxylase – Present in mito- >> Fructose 16 bisphosphatase is the rate
chondria limiting enzyme of gluconeogenesis
• Phosphoenol pyruvate carboxykinase
• Glucose 6 phosphatase – Present in En-
doplasmic reticulum
>> Glucose 6 phosphatase is present in liver
kidney making them gluconeogenic
>> 11 high energy phosphates are required
for converting two molecules of pyruvate to
1 molecule of glucose
• Pyruvate carboxylase – uses 2 high ener-
gy phosphates
75
76
GLUCONEOGENIC SUBSTRATES
STEPS OF GLUCONEOGENESIS
@dr.c.shanmugapriya
Dr. C. Shanmugapriya BIOCHEMISTRY
>>>
Regulation of Glycolysis and

Gluconeogenesis
• Common allosteric regulator is Fructose • Fructose 2,6 bisphosphate is a product
2,6 bisphosphate of a tandem enzyme – PFK-2
• Fructose 2,6 bisphosphate is a stimula- • PFK-2 has both PFK2 activity and Fruc-
tor of glycolysis (PFK-1) tose 2,6 bisphosphatase activity
• Fructose 2,6 bisphosphate is an inhibitor • Fructose 2,6 bisphosphatase activity is
of gluconeogenesis (Fructose 1,6 bis- stimulated by phosphorylation
phosphatase)
Types of Glut Transporters

S.No GLUT transporter Location
1 GLUT 1
2 GLUT 2
3 GLUT 3
4 GLUT 24
5 GLUT 5
77
S.No GLUT transporter Location
1 GLUT 1 Neurons, RBCs and

Placenta
2 GLUT 2 Enterocytes, Hepatocytes,

Pancreatic β cells, PCT
3 GLUT 3 Neurons, RBCs and

Placenta
4 GLUT 4 Skeletal muscle, Cardiac

muscle, adipose tissue
5 GLUT 5 Fructose absorption
Alcohol metabolism
• Alcohol is metabolized by alcohol dehy- • The high NADH/NAD ratio shifts the
drogenase to form an aldehyde. Alde- equilibrium between pyruvate and
hyde dehydrogenase converts aldehyde lactate to lactate formation, hence it
to acid causes lactic acidosis and low pyru-
• Both the dehydrogenases use NAD and vate levels. It also shifts the equilibrium
form NADH. Hence the metabolic status between malate and oxaloacetate to
of a chronic alcoholic is high NADH:NAD malate formation. This causes low ox-
ratio aloacetate levels. Low pyruvate and
low oxaloacetate decreases the rate of
• NADH gets converted to ATP gluconeogenesis. This causes repeated
• Hence, alcohol is a source of empty episodes of hypoglycemia
calorie (calorie without essential micro- • High energy causes anabolism, Hence
nutrient) they present with weight gain and fatty
• This causes micronutrient deficiencies liver
including but not limited to thiamine
ALCOHOL METABOLISM
R- CH2OH → RCHO → RCOOH
ALCOHOL DEHYDROGENASE ALDEHYDE DEHYDROGENASE
NADH NADH
ATP ATP
78 >>>
ALCOHOLISM AND HYPOGLYCEMIA
PYRUVATE LACTATE LOW PYRUVATE LACTATE
NADH
LOW
OXALOACETATE MALATE MALATE
OXALOACETATE
MCQS
1. Which of the following is a glucogenic
B. Glycerol
C. Lactate
organ? D. Alanine
5.Following an early morning run, a 29

A. Liver
B. Skeletal Muscle
C. Lens year old man consumes a carbohydrate
D. Cardiac muscle rich south Indian breakfast. Which of the
2. Number of high energy phosphates

following will most likely be activated in his
liver after breakfast?
required when pyruvate is used as a gluco- A. Cytoplasmic PEPCK
neogenic substrate? B. Membrane GLUT4 transporter
A. 6 C. Cytoplasmic PFK1
B. 4 D. Cytoplasmic Glycogen Phosphorylase
6. A 55 year old alcoholic was brought

C. 11
D. 10
3.The rate limiting enzyme of gluconeo-

to the emergency department by his
friends, During their usual hangout at the
genesis? local bar, he had passed out and they were
A. Glucose 6 phosphatase unable to revive him. On admission, his
B. Pyruvate carboxylase blood glucose was 55mg/dL.Question 1:
1.A; 2.C; 3. D; 4.A.5.C; 6.C
C. PEPCK Which of the following is the explanation for

D. Fructose 1,6 bisphosphatase hypoglycemia in him?
4.Which of the following is not a gluco-

A. Thiamine deficiency
B. High NAD
genic substrate? C. Low Pyruvate
A. Acetyl CoA D. High Oxaloacetate
79
PYQ Discussion
I. Thiamine deficiency causes lactic acido-

sis because of inhibition of?
A. PDH
B. Pyruvate decarboxlase MSUD RESPONDS
C. PEPCK TO THIAMINE
D. Transketolase SUPPLEMENTATION
THIAMINE DEPENDENT ENZYMES

• PDH • BCKADH
• Alphaketoglutarate de- • Transketolase

hydrogenase
THIAMINE DEFICIENCY
• In thiamine deficiency, PDH complex is » PDH complex is essential for aero-
inactive. There are two effects bic utilisation of glucose. Neurons
» Pyruvate accumulates and is con- use glucose aerobically, hence in
verted to lactate. This is why thi- thiamine deficiency, neuron’s metab-
amine deficieency causes lactic olism gets affected. This causes dry
acidosis. Lactic acid is a vasodilator beriberi
and this causes hypotension and • Transketolase – Thiamine deficiency is
tachycardia – wet beriberi detected by estimating RBC transketo-
lase activity
II. A person has a history of alcoholism

for 20 years. He avoids alcohol for 2 days
and presents with agitation, global confu-
sion, disorientation, hallucination, fever, high
BP, diaphoresis and autonomic hyperactivi-
ty. The most common cause is
A. Delirium tremens
B. Wernicke’s encephalopathy
C. Korsakoff syndrome
I.A;II.A
D. Pancreatitis
80 >>>
ALCOHOLISM & THIAMINE

DEFICIENCY
CAUSES: EFFECTS:
• As alcohol is a source of empty calorie • Acute thiamine deficiency presents as
(high energy no essential micronutri- Wernicke’s encephalopathy. The pres-
ents), the person skips meals and hence entations of Wernicke’s encephalopathy
is prone to develop deficiencies of all are Global confusion, ophthalmoplegia
micronutrients and Ataxia (GOA)
• Alcohol interferes with thiamine absorp- • Chronic thiamine deficiency presents as
tion Korsakoff syndrome. It presents as ret-
• Alcohol interferes with magnesium rograde amnesia, anterograde amnesia
absorption. Magnesium is necessary for and confabulation or honest lies. Senso-
the conversion of thiamine to thiamine ry agnosia is also a feature
pyrophosphate – the active form of
thiamine
DELERIUM TREMENS
• It is a feature of alcohol withdrawl. It • It also presents with physical symptoms
presents between the 2 and the 10
nd th
and signs like High body temperature,
day of alcohol withdrawal. High HR, High BP, shaking because of
• It presents as Global confusion, Halluci- autonomic hyepractivity
nation, Nightmares
III. Drug of choice in mountain sickness

A. Thiazides
B. Frusemide
C. Acetazolamide
III.C
D. Spironolactone
Mountain Sickness
• It is associated with high altitude. As washes away carbon dioxide, this results
the partial pressure of oxygen in the in Respiratory alkalosis.
atmosphere is low in high altitudes, the • Respiratory alkalosis depresses the res-
personhyperventilates and the person piratory centre, as the central chemore-
ceptors present in medulla respond only
81
to an increase in hydrogen ion concen- • Acetazolamide is a carbonic anhydrase

tration in the CSF inhibitor, which inhibits bicarbonate
• To counteract this or to treat this, we reabsorption in PCT. Loss of bicarbonate
have to induce metabolic acidosis. This in PCT causes metabolic acidosis
is achieved by acetazolamide
ACETAZOLAMIDE
Oxyhemoglobin Dissociation Curve

• It is a curve drawn » High P50
with partial pressure » Low affinity
of oxygen along x » Effective deoxy-
% saturation
axis and percentage genation to the

saturation along y tissues
axis » Hypoxia
• The position of the » Anemia
curve determines » Acidosis
P50 » Acclimatization
• P50 is the partial • Shift of the curve to
pressure of oxygen the left:
at which it is 50% » Low P50
saturated. Low P50 » High Affinitt
means higher affinity » Alkalosis
PO2
of Hemoglobin for » Dexrease in body
oxygen temperature
• Shift of the curve to » Fetol Hemoglobin
the right: » CO poisoning
82 >>>
IV. Which of the following shifts the

given curve to left?
A. Hypoxia
B. Acidosis
C. Anemia
IV.D
D. Decrease in body temperature
RATE LIMITING ENZYMES OF CARBOHYDRATE

METABOLISM
S.No PATHWAY RATE LIMITING ENZYME
1 Glycolysis
2 Citric acid cycle
3 Glycogen synthesis
4 Glycogenolysis
5 Gluconeogenesis
6 HMP shunt
83
1 Glycolysis PFK-1
2 Citric acid cycle ICDH
3 Glycogen synthesis Glycogen Synthase
4 Glycogenolysis Glycogen phosphorylase
5 Gluconeogenesis Fructose 1,6 bisphosphatase
6 HMP shunt Glucose 6 phosphate dehydrogenase
Energetics
Number of ATPs from Number of ATPs from

S.N o
Pathway 1 molecule of free 1 molecule of glucose
glucose from glycogenolysis
1 Aerobic glycolysis
2 Anaerobic glycolysis
3 Complete oxidation
4 Glycogen synthesis
5 Gluconeogenesis
Number of ATPs from Number of ATPs from

S.N o
Pathway 1 molecule of free 1 molecule of glucose
glucose from glycogenolysis
1 Aerobic glycolysis 7 8
2 Anaerobic glycolysis 2 3
3 Complete oxidation 32 33
4 Glycogen synthesis -3/ every molecule of glucose
5 Gluconeogenesis 11
84 >>>
VI. The additive that is used in the given

RSS
blood collection tube is? Red
Silica
A. Sodium Fluride
B. K2EDTA Serum
C. Heparin
VI.C
D. Silica CLOT ACTIVATOR
Types of blood collection tubes
• Sodium Fluride and • Citrated tube • K2 EDTA Tube

Potassium Oxalate • Coagulation assays • CBC
• Glucose estimation • HbA1C
• Ammonia
• ACTH
• Heparinised tube • Gel and clot tubes

• Stat Biochemical analysis • Enables sample storage
• Cytogenetics
85
Enzymes
Introduction
Enzymes are biocatalysts. Bio because RNA and snRNAs. They increase the rate of
they are made up of biologically important the chemical reaction and they are neither
macromolecules. They are all proteins with utilized nor generated during the process.
the few exceptions which are RNAs. Such They increase the rate of a chemical reac-
RNAs with enzymatic activity are called as tion by reducing the activation energy of a
ribozymes. Example for ribozymes are 28sr- chemical reaction.
Classification of enzymes
• OXIDOREDUCTASES • LYASES • TRANSLOCASES
• TRANSFERASES • ISOMERASES
• HYDROLASES • LIGASES
To avoid ambiguity, the International Un- denotes the class, the second digit is the
ion of Biochemistry and Molecular Biology subclass and third digit stands for the group
proposed that enzymes be classified into and the fourth digit is the number assigned
classes, subclasses and groups and that to a particular enzyme in the group list.
in each group, enzymes are arranged in a For example, alcohol dehydrogenase is
sequence. This paved the way for every named as EC.1.1.1.1, which means alcohol
enzyme getting a unique name with EC dehydrogenase belongs to the first of the
(meaning enzyme class) as a prefix followed six classes of enzymes, first subclass, first
by 4 digits separated by dots. The first digit group, first enzyme.
CLASSES OF ENZYMES
There are seven classes of 2. TRANSFERASES 5. ISOMERASES

enzymes: 3. HYDROLASES 6. LIGASES
1. OXIDOREDUCTASES 4. LYASES 7. TRANSLOCASES
or the coenzyme gets reduced or oxidised

>> CLASS 1: OXIDOREDUCTASES reciprocally.
Oxidoreductases catalyse oxidation or re- Oxidoreductases are classified into 21 sub-
duction reactions of the primary substrate, classes. Some of those are,
and hence either the secondary substrate
1. Dehydrogenases
87
2. Oxidases
2. Peptidases
3. Monoxygenases / Hydroxylases
3. Esterases
4. Dioxygenases
4. Phosphatases
5. Peroxidases
5. Phospholipases
6. Reductases
6. Glycosidases
7. Catalases
7. Ribonucleases
Examples: Alcohol dehydrogenase, Lactate
Examples include, aminopeptidase. Car-
dehydrogenase, L aminoacid oxidase, Ty-
boxypeptidase A, carboxypeptidase B, Cho-
rosine hydroxylase, p- hydroxyphenylpyru-
lesterol ester hydrolase, Lipoprotein lipase,
vate dioxygenase, Glutathione peroxidase,
Phospholipase.
Glutathione reductase, Catalase
>>CLASS 4: LYASES:
>>CLASS 2: TRANSFERASES:
Lyases cleave a covalent linkage without
Transferases transfer a group from one
adding water. Hence, they serve to break
substrate to the other substrate. They are
C-N /C-C linkage without adding water. As
classified into 10 subclasses. Some of those

all chemical reactions are reversible, they
include
serve to form C-N/ C-C linkage as well.
1. Kinases /Phosphotransferases (as Hence lyases also include synthases.
kinases transfer phosphate group from
Some subclasses of lyases include
one substrate to another)
2. Transketolases 1. Decarboxylases
3. Transaldolase 2. Hydratase
4. Acyltransferase 3. Dehydratase
5. Glucosyltransferase – glycogen 4. Aldolase
synthase (Glycogen synthase tranfers 5. Lyases
glucose from UDP glucose to glycogen 6. Synthases
primer) Examples include ALA synthase of heme
synthesis, Hydratase of fatty acid synthesis,
Examples include hexokinase, transke-
Aldolase A of glycolysis, ATP citrate lyase of
tolase and transaldolase of HMP shunt,
fatty acid synthesis
Glycerol 3 phosphate acyl transferase and
glycogen synthase
>>CLASS 5: ISOMERASES
>>CLASS 3: HYDROLASES: Isomerases convert one isomer into anoth-

er. In short, the molecular formula of the
Hydrolases cleave a covalent linkage by
substrate and the product are the same.
addition of water. Hydrolytic cleavage can
break only ester or ether linkages. Exam- Few subclasses of isomerases include,
ples of ester linkages include phosphate or 1. Isomerases
peptide linkages. Examples of ether linkag- 2. Racemase
es including glycosidic linkages found in 3. Epimerase
carbohydrates and in nucleotides. 4. Mutase – Mutase is an enzyme which
Some subclasses of hydrolases include, shifts a group from one carbon atom to
another.
1. Amidases
88 >>>
Examples include, phosphotriose isomer- >>CLASS 6: LIGASES

ase of glycolysis, MethylmalonylCoA Ligases are enzymes which synthesises a
racemase of propionyl CoA metabolism, covalent linkage using ATP as a source of
Phosphoglycerate mutase of glycolysis, energy
bisphosphoglycerate mutase of Rapaport
Few sublasses include
Leubering shunt
1. Synthetases
2. Ligases
3. Carboxylases
Differences between
hydrolases and lyases
S.No HYDROLASES LYASES
S.No HYDROLASES LYASES
1 Addition of water Without adding water
2 Ester and ether linkages C-N or C-C

are broken
3 Examples – Decarboxylase,
Phosphatases, amidases, hydratase, dehydratase,
peptidases, proteases, lyases and synthase
phospholipases,
glycosidases,
ribonucleases and
deoxyribonuclease
89
Differences between
Lyases and Ligases
S.No LYASES LIGASES
3
S.No LYASES LIGASES
1 Does not need ATP Needs ATP
2 Synthase Synthetases
3 Decarboxylases Carboxylases
Points to remember
• Hydroxylase is a monooxygenase • Hydratase and dehydratases are lyases
• Catalase is an oxidoreductase • Synthases don’t use ATP
• Glycogen synthase is a transferase • Decarboxylases are lyases and
• Hydrolases add water and lyases don’t carboxylases are ligases
MCQS
1. Glycogen Phosphoryl- 2. Glycogen synthase is a, 3.ALA synthase is an
ase belongs to example of
A. Ligase
A. Class 6 B. Lyase A. Lyase
1.D; 2.C; 3. A;
B. Class 4 C. Transferase B. Ligase

C. Class 3 D. Hydrolase C. Transferase
D. Class 2 D. Hydrolase
90 >>>
Non protein part of

an enzyme
The protein part of an enzyme is called as zyme. The non protein parts of an enzyme
apoenzyme. The protein and the non protein are of three types -PROSTHETIC GROUPS/
part of an enzyme is called as a holoen- COFACTORS /COENZYMES
Prosthetic groups
If the non protein part of an • FAD in succinate dehy-
enzyme is in a tight, sta- drogenase
ble incorporation with the • Magnesium in kinases
protein part, it called as a • Mn in SOD
prosthetic group. Examples • Copper in Tyrosinase
- PLP, TPP, Biotin, FAD, FMN and SOD
and Mg, Mn, Co, Cu, Zn • Zinc in carbonic anhy-
• PLP in transaminases drase, alkaline phos-
and decarboxylases phatase, alcohol dehy-
• TPP in PDH complex drogenase
• Biotin in carboxylases
Cofactors
• If the non protein part protein part, it is called
of an enzyme is in a as cofactors
transient, dissociable • Examples: Ca, Mg de-
incorporation with the pendent kinases
Coenzymes
• If the non protein part • Examples:
of an enzyme acts as a • Methyl group – folates,
Recyclable shuttle/ group • acyl group – CoA,
transfer agent, it is called • Oligosaccharides -
as coenzymes Dolichol
91
MCQS
1. PLP is a, 3.Cobalamine in methionine synthase
acts as ,
A. Prosthetic group
B. Cofactor A. Prosthetic group
C. Apoenzyme B. Cofactor
D. Holoenzyme C. Apoenzyme
2. Folate derivatives act as,

D. Coenzymes
4.Dolichol acts as,

A. Prosthetic group
1.A; 2.C; 3. A; 4.D

B. Cofactor A. Prosthetic group
C. Apoenzyme B. Cofactor
D. Coenzymes C. Apoenzyme
D. Coenzyme
Enzyme Kinetics and

Enzyme Inhibition
Enzyme Kinetics
• Most of the enzymes follow saturation • Vmax is the velocity of the enzyme cata-
kinetics. lysed chemical reaction when all the avail-
• When there is an increase in the substrate able enzyme’s substrate binding sites are
concentration, the rate of the enzyme cat- saturated with the substrate. Vmax is never
alysed chemical reaction increases propor- measurable directly as enzymes are cata-
tionately till a point, beyond which, inspite lysts and are required in minute concentra-
of there being an increase in the substrate tion. So enzymes never get saturated.
concentration, velocity does not increase, • Km is the substrate concentration at half
as all enzyme’s substrate binding sites get maximal velocity.
saturated with the substrate. • Lower Km signifies higher affinity
92 >>>
MICHAELIS: MENTON EQUATION &

SIGNIFICANCE OF KM
V = Vmax . [S] >> When u substitute V = Vmax/2

Km + [S]
• Vmax = Vmax [S]
Where v is the velocity of the enzyme • 2 km + [S]
catalysed chemical reaction for a given • Km + [S] = 2[S]
substrate concentration [S] • Km = [S]
LINEWEAVER BURKE PLOT OR

DOUBLE RECIPROCAL PLOT
• As Vmax can not be measured directly,

km can not be measured directly This is in the form of equation of a straight
• Lineweaver Burke plot or double line, y=mx +c
reciprocal plot helps in measuring Vmax
and Km • Hence if a graph is drawn with 1/v along
• When you take reciprocals of Michelis y axis and 1/[S] along x axis, we get a
menton equation it is, straight line
1 = Km . 1 + 1 • The straight line cuts the y axis at 1/
v Vmax [S] Vmax vmax and it cuts the x axis at -1/Km
93
USE 1: To identify the type of enzyme inhibition

• To know the type of presence of and in the • Depending upon the
enzyme inhibition we absence of inhibitor changes on Vmax and
subject the enzyme to its • We calculate the Km and Km, the type of inhibition
activity in the presence Vmax of the enzyme both can be identified.
of varous substrate con- in the presence of and
centration both in the absence of inhibitor
Types of inhibition
» Reversible Inhibition » Irreversible inhibition
Types of reversible inhibition

>> Competitive inhibition: The inhibitor is >> Mixed inhibition: The inhibitor binds
a substrate analog and it competes with the to both the free enzyme and the enzyme
substrate to bind to the substrate binding substrate complex and the product is not
site of the free enzyme. formed
>> Uncompetitive inhibition: The inhibi- • A special type of mixed inhibition is non
tor binds to the enzyme substrate complex competitive inhibition, where the inhib-
and does not allow the complex to be con- itor binds to both the free enzyme and
verted to product the enzyme substrate complex with the
same affinity
Effects of competitive Inhibition

• As a competitive inhibitor reduces the
COMPETITIVE INHIBITION
probability of a substrate binding to
the enzyme, increasing the substrate
concentration increases the probability
that the substrate overcomes the
inhibition by the competitive inhibitor
and hence the impact of inhibition is
overcome.
• As the inhibition is overcome, Vmax
or the maximum velocity achievable
remains normal
• To achieve Vmax, as we have to increase
the substrate concentration, Km is
increased
94 >>>
Effects of Uncompetitive Inhibition

• As an uncompetitive inhibitor binds to
UNCOMPETITIVE INHIBITION the enzyme substrate complex, how
much ever the substrate concentration
is increased, the product is not formed.
• As the inhibition is not overcome, Vmax or
the maximum velocity achievable is low
• As the inhibitor binds to enzyme
substrate complex, the propensity to
form more enzyme substrate complex
increases and hence more enzymes
bind to substrate now, increasing
indirectly the affinity between enzyme
and substrate. Hence the Km is low
• Both Km and Vmax are low
Effects of Mixed Inhibition

• As a mixed inhibitor binds to both the
free enzyme and the enzyme substrate
complex, how much ever the substrate
MIXED INHIBITION concentration is increased, the product
is not formed.
• As the inhibition is not overcome, Vmax
or the maximum velocity achievable is
low
• As the inhibitor binds to free enzyme
the probability that the enzyme
substrate complex is formed is
reduced. So affinity between enzyme
and substrate is reduced. Hence the Km
is high
• Km is high and Vmax is low
95
Effects of Non Competitive Inhibition
• A non competitive inhibitor binds to

both the free enzyme and the enzyme
PURE NON-COMPETITIVE
INHIBITION substrate complex with the same affinity
• Hence Km is normal and Vmax is low
Effects of inhibitor ON Km and Vmax

S.No TYPE OF INHIBITION Vmax Km
1 Competitive Inhibition
2 Uncompetitive Inhibition
3 Mixed Inhibition
4 Non Competitive Inhibition
96 >>>
S.No TYPE OF INHIBITION Vmax Km
1 Competitive Inhibition Normal High
2 Uncompetitive Inhibition Low Low
3 Mixed Inhibition Low High
4 Non Competitive Inhibition Low Normal
Suicidal Inhibition
• It is a type of irreversible inhibition which inhibits the same enzyme by
• The inhibitor is a substrate analogue which it is activated, hence the name
and hence binds to the active site of the suicidal inhibition
enzyme • Furoacetate is a suicidal inhibitor of
• The enzyme catalyses the conversion of aconitase
the substrate to a reactive intermediate,
97
MCQS
1. Michaelis menton equation is used for 3.True about double reciprocal plot is,
enzymes following, A. 1/v is plotted along x axis
A. Linear kinetics B. 1/[S] is plotted along y axis
B. Saturation kinetics C. 1/vmax is the x intercept
C. Sigmoidal kinetics D. -1/km is the x intercept
4.True regarding competitive inhibition of

D. Hyperbolic kinetics
2. Equation used to study the kinetics of an enzyme is,

enzymes following sigmoidal kinetics is, A. Km is increased
A. Lineweaver Burke equation B. km is unaltered
C. Km is decreased
B. Michaelis menton equation

C. Hill’s equation D. Vmax is decreased
D. Dixon equation
5. What kind of inhibition is this?

Linear kinetics
A. Competitive inhibition
B. Uncompetitive inhibition
C. Non competitive inhibition
D. Mixed inibition
6. An enzyme was mixed with 4mM sub- SOLUTION
strate. The initial rate of product formation

was 25% of Vmax. The Km of the enzyme is V = Vmax [S]
A. 2mM km + [S]
B. 4mM When [S] = 4mM, V = Vmax/4
C. 9mM Vmax = Vmax x 4
D. 12mM 4 Km + 4
98 >>>
7. An enzyme-catalyzed reaction was 8.Suicidal inhibition, true statement is/

carried out with the initial substrate con- are?
centration 1,000 times greater than the Km
for that substrate. After 9 minutes, 1% of A. Substrate analogue that binds to some
the substrate had been converted to the site of enzymes
product, and the amount of product was B. They react with the enzymatic site and
12 mmol. If, one-third as much enzyme and irreversibly inhibit it
twice as much of the substrate is combined, C. The inhibitor does not react outside the
how long it would take for the same amount active site
(12 mmol) of product to be formed? D. The inhibitor forms a product with the
A. 13.5 mins help of the enzyme and the product
B. 27 mins inhibits it
C. 3. 8 mins
D. 4. 9 mins
DECIPHERING THE QUESTION
• When [S] = 1000Km, V = 12mmol in • When [S] = 2000 Km and [E] =

9 minutes 0.33[E], how long does it take for
the same 12mmol of products to be
formed?
SOLUTION
• When [S] = Km, V = 0.5Vmax spective of whether it is 1000Km

• When [S] = 10 Km, V = 0.9Vmax or 2000km, velocity is 12mmol in 9
minutes
• When [S] = 100km, V = 0.99 Vmax
1.B; 2.C; 3.D; 4.A; 5.B; 6.D; 7.B; 8.A,B&D.
• Beyond 10km, the enzyme concen-

• When [S] = 1000km, V = tration is the deciding factor for
0.999Vmax the velocity of enzyme catalysed
• Hence, as [S] increases from 0 to chemical reaction. As the enzyme
10km, increase in substrate con- concentration is 1/3rd, time taken
centration increases the velocity. will be tripled. Hence now its 27
Beyong 10km, increase in the minutes
substrate concentration, can not
increase the velocity. Hence irre-
99
Electron
Transport Chain
Introduction
Electron Transport Chain
• It is a chain of complexes that is present • To allow electrons from NADH to pass

along the inner side of inner mitochon- through ETC, first NADH has to be trans-
drial membrane ported from cytoplasm to mitochondria
• It is through this chain of complexes that • The only source of NADH in cytoplasm
we move the electrons that are obtained is glycolysis – Glyceraldehyde 3 phos-
by oxidizing NADH and FADH2 to pass phate dehydrogenase generates 2
through, which in a complex way gener- NADH from every molecule of glucose
ates energy. This energy is used for the that enters into glycolysis
phosphorylation of ADP to form ATP • Electrons of these two NADH enter mito-
• ETC is a tool of oxidative phosphoryla- chondria by malate shuttle or by Glycer-
tion ol phosphate shuttle
Shuttles
Malate Shuttle & Glycerol

Phosphate Shuttle
101
GLYCEROL PHOSPHATE
S.No PROPERTY MALATE SHUTTLE
SHUTTLE
1 Cells All White muscle fibres and

neurons
2 NADH electron Malate Glycerol Phosphate

transported as
3 Enzymes and Cytoplasmic Malate Cytoplasmic

Coenzymes used Dehydrogenase – uses NAD Glycerol phosphate
Mitochodnrial Malate dehydrogenase: uses NAD
dehydroegenase – Uses NAD Mitochodnrial Glycerol
phosphate dehydrogenase
– Uses FADH2
4 NADH 2.5 ATP 1.5ATP

Aerobic 7 5
glycolysis
Eneergectics
Anaerobic 3 2
glycolysis
Complete 32 30
oxidation of 1
mole of glucise
Malate Shuttle & Glycerol Phosphate

shuttle
• It is a chain of complexes that is present • To allow electrons from NADH to pass
along the inner side of inner mitochon- through ETC, first NADH has to be trans-
drial membrane ported from cytoplasm to mitochondria
• It is through this chain of complexes that • The only source of NADH in cytoplasm is
we move the electrons that are obtained glycolysis – Glyceraldehyde 3 phosphate
by oxidizing NADH and FADH2 to pass dehydrogenase generates 2 NADH from
through, which in a complex way gener- every molecule of glucose that enters
ates energy. This energy is used for the into glycolysis
phosphorylation of ADP to form ATP • Electrons of these two NADH enter mito-
• ETC is a tool of oxidative phosphoryla- chondria by malate shuttle or by Glycer-
tion ol phosphate shuttle
102 >>>
Mitchelle’s
Chemiosmotic theory
Facts about ETC: Chemiosmotic Theory
• All complexes of ETC are arranged in • After hydrogen ions accumulate outside
an increasing order of redox potential/ the inner mitochondrial membrane, they
increasing order of state of oxidation/ find their way to the mitochondrial matrix
decreasing order of energy level by going through F0 Component of ETC
• When electrons move from one complex • As here it is a movement of hydrogen ion,
to another, they move from a complex of from a region of higher to lowe concen-
higher energy to a complex of lower ener- tration, energy is liberated. The energy is
gy. This causes energy to be liberated used for generating ATP by F1 component
• The energy is used to pump hydrogen of ETC or ATP synthase
ions from the matrix to outside the inner
mitochondrial membrane
Facts about ETC: Calculation

• When electrons move from complex I to Q, • When FADH2 enters ETC, 6 Hydrogen ions
4 hydrogen ions are translocated are translocated
• From Q to III, 4 hydrogen ions are translo- • 4 hydrogen ions passing through ATP
cated synthase complex causes 1 ATP to be
• From III to IV, 2 hydrogen ions are translo- generated
cated • Hence NADH generates 2.5 ATPs, FADH2
• When NADH enters ETC, 10 hydrogen ions generates 1.5 ATPs
are translocated
FADH2 linked Dehydrogenases

• Succinate Dehydrogenase of TCA cycle • Mitochondrial Glycerol 3 phosphate dehy-
• Acyl CoA dehydrogenase of Fatty acid drogenase
oxidation
103
Complexes of ETC
COMPLEX II
SUCCINATE
DEHYDROGENASE
COMPLEX I COMPLEX COMPLEX

III IV
NADH LINKED CYT b & CYT a &

Q C O2
DEHYDROGENASE c1 a3
ACYL CoA
DEHYDROGENASE/ MT
GLYCEROL 3 PHOSPHATE
DEHYDROGENASE
MITCHELLE’S CHEMIOSMOTIC THEORY
COMPLEX II
SUCCINATE
DEHYDROGENASE

III IV

Q C O2
DEHYDROGENASE c1 a3
REDOX POTENTIAL/
STATE OF OXIDATION
ENERGY LEVEL
104 >>>
PROTON PUMP COUPLED TO PHOSPHORYLATION
TRANSLOCATION OF 10 H+ WHEN NADH ENTERS ETC
10 H+
COMPLEX II
SUCCINATE
DEHYDROGENASE

III IV

Q C O2
DEHYDROGENASE c1 a3
4 4 2
REDOX POTENTIAL/
STATE OF OXIDATION
ENERGY LEVEL
105
TRANSLOCATION OF 6 H+ WHEN FADH2 ENTERS ETC
10 H+
COMPLEX II
SUCCINATE 6 H+
DEHYDROGENASE

III IV

Q C O2
DEHYDROGENASE c1 a3
4 4 2
REDOX POTENTIAL/
STATE OF OXIDATION
ENERGY LEVEL
Inhibitors
Inhibitors of ETC
COMPLEX II
SUCCINATE
DEHYDROGENASE

III IV

Q C O2
DEHYDROGENASE c1 a3
106 >>>
Complex I to Q: Amobarbital, Piericidin A and Rotenone
Complex II to Q: TTFA & Carboxin
Complex III to C: BAL & Antimycin
Complex IV to Oxygen: H2S, Cyanide and CO
Uncoupler: 2,4 Dinitrophenol, Valinomycin, Nalinomycin and Nigericin
ATP synthase: Oligomycin
Uncouplers
• Electron transport chain couples oxidation of fuels with
phosphorylation of ADP to form ATP
• Uncouplers uncouple oxidation from phosphorylation
• Oxidation of fuel happens continuously without phos-
phorylation
• Oxidation of fuels happens continuously and energy is
liberated continuously but the liberation of energy is not
coupled to ATP production
• Hence energy is liberated out as heat
• Effects of uncouplers:
» No ATP production
» Excess heat production
» For want of ATP more fuel gets oxidized and hence
there is increase in the rate of oxidation of fuels
• Mechanism of action:
• All uncouplers are ionophores or ion channels and they
get inserted into the inner mitochondrial membrane.
Through these channels, hydrogen ions translocate. As
hydreogen ions do not go through F0 component ATP is
not generated. However as hydrogen ion moves from a
region of hgher to lower concentration, energy is liber-
ated out as heat
• Examples of uncouplers:
» Physiological – Brown adipose tissue and throxine
» Artificial uncoupler – 2,4 Dinitrophenol, Valinomycin,
Nalinomycin and Nigericin
107
ONE LINERS
• The ETC Complex with copper is Complex IV
• Mobile complexes are Q and Cytochrome C
• The complex in ETC without Heme is Q
• The final acceptor of electron in ETC is Oxygen
• Complex II is Succinate Dehydrogenase
MCQS
1. According to Mitchelle’s chemiosmotic 3. Antimycin inhibits the electron trans-

theory, which of the following is true? port at which level:
A. They are arranged in a decreasing order A. Complex I to Q

of redox potential B. Complex II to Q
B. They are arranged in a decreasing order C. Complex III to C
of ability to get reduced D. Complex IV to Oxygen
C. They are arranged in a decreasing order
of energy level
D. They are randomly arranged
2. The number of H+ translocated, when

FADH2 enters ETC:
A. 10
B. 6
C. 3
D. 2
108 >>>
INTEGRATED CASE
BASED MCQ
4. An obese woman, who is desperate to 5. Marilyn Manroe, the famous American
lose weight takes an online weight reduction model, actress and singer was suffering
pill and ends up in hyperthermia. A para- from insomnia. Her friend introduced her to
medic checks the content of the pill and a street drug “Lilly 33s.”, which is amobar-
finds valinomycin as one of the components. bital!! She died at the age of 36, following
Mechanism of action of valinomycin is intake of few “Lillies”!! Amobarbital acts by
inhibiting electron transport from
A. It is an uncoupler
B. It inhibits ATP synthase A. Complex I to Q
C. It inhibits ATP/ADP translocator B. Complex II to Q
D. It inhibits electron transport between C. Complex III to C
Complex I and Q D. Complex IV to Oxygen
IMAGE BASED MCQS
6. INHIBITOR X acts on a trans-

porter marked in the image. Inhibitor
1.C; 2.A; 3.C; 4.a; 5.A; 6.D.
A is
A. Hydrogen Sulphide
B. Oligomycin
C. 2,4 Dinitrophenol
D. Atractyloside
109
Heme Synthesis
and Porphyrias
Introduction
• Heme is a metal containing protoporphy- related manifestations – erythrodontia,
rin ring portwine urine
• The metal in the centre is iron in hemo- • Absorption of 400nm photon is responsi-
globin and myoglobin, copper in cy- ble for soret band
tochrome oxidase, magnesium in chloro- • As it absorbs photon of higher energy
phyll (shorter wavelength 400nm) and emits
• Protoporphyrin rings have four pyrrole photon of lesser energy (longer wave-
rings connected by methenyl bridges length 600nm), the energy difference is
with conjugated double bonds emitted as free energy. The free energy
• These double bonds absorb light at excites oxygen to form superoxide rad-
400nm and emit light at 600nm. 600nm ical and this is responsible for photooxi-
is responsible for red fluorescence and dative damage caused in porphyrias
PORPHYRINOGEN RING UROPORPHYRINOGEN RING
UROPORPHYRINOGEN I & III COPROPORPHYRINOGEN I & III
111
Heme synthesis
Conversion of Uroporphyrinogen to
Coproporphyrinogen
DECARBOXYLASE
Conversion of Coproporphyrinogen
to protoporphyrinogen
OXIDASE
112 >>>
Side chains in porphyrins

• Uroporphyrinogen – • Coproporphyrinogen – • Protoporphyrinogen –
Acetyl Propionyl Methyl Propionyl Methyl Vinyl
Steps of Heme Synthesis
• Involves 8 steps • In the presence of Uroporphyrinogen

• 4 steps in mitochondria and 4 steps in III synthase, Hydroxymethylbilane forms
cytoplasm Uroporphyrinogen III
• In the first step, glycine reacts with suc- • Uroporphyrinogen I and III in the pres-
cinyl CoA in mitochondria in the presence of Uroporphyrinogen decar-
ence of ALA synthase to form ALA. It boxylase form Coproporphyrinogen I
is a decarboxylation step. Hence it uses and III
PLP as a coenzyme • Coproporphyinogen III enters mitochon-
• ALA reaches cytoplasm. 2 molecules of dria and in the presence of Copropor-
ALA condense in the presence of ALA phyrinogen oxidase, it is converted to
dehydratase to form Porphobilinogen. Protoporphyrinogen
This step uses Zinc. • Protoporphyrinogen is oxidized in the
• 4 Porphobilinogen condense by the re- presence of protoporphyinogen oxi-
moval of four ammonia in the presence dase to form protoporphyrin
of PBG deaminase to form Hydroxym- • Ferrochelatase chelates iron into pro-
ethilbilane toporphyrin to form heme
• Hydroxymethylbilane spontaneously
cyclises to form Uroporphyrinogen I
113
BIOCHEMISTRY I A COMPLETE COURSE I Part 01 @dr.c.shanmugapriya Dr. C. Shanmugapriya BIOCHEMISTRY
Enzymes of Heme Synthesis
• MITOCHONDRIA • FERROCHELATASE PHYRINOGEN I SYN-

• ALA SYNTHASE • CYTOPLASM THASE
• COPROPORPHYRINOGEN • ALA DEHYDRATASE • UROPORPHYRINIGEN III
OXIDASE • PBG DEAMINASE/ HMB SYNTHASE
• PROTOPORPHYRINIGEN SYNTHASE/ UROPOR- • FERROCHELATASE
OXIDASE
Regulation of Heme Synthesis
• ALA Synthase I in liver • ALA Synthase II in RBC precursors stim-

• ALA Synthase II in RBC precursors ulated based on iron availability and rate
• ALA Synthase I in liver inhibited by of erythropoiesis
heme • ALA dehydratase is inhibited by Copro-
porphyrinogen and Protoporphyrinogen
114 >>>
Heme Synthesis
Disorders
Name of the Clinical and Biochemical

S.No Enzyme Defect
disorder Features
1
115
Name of the Clinical and Biochemical

S.No Enzyme Defect
disorder Features
1 ALA Synthase II X linked Anemia with iron laden

sideroblastic anemia mitochondria surrounding nuclei
in RBC precursors
2 ALA Dehydratase ALA Dehydratase Neuropsychiatric manifestations

Deficient Porphyria with periodic aggressive behaviour
and sensory neuropathy. ALA is
elevated in urine
3 PBG Deaminase or Acute Intermittent Neuropsychiatric manifestations

Hydroxymethilbilane Porphyria with periodic aggressive behaviour
synthase or and sensory neuropathy. ALA and
Uroporphyrinogen I PBG are elevated in urine
synthase
4 Uroporphyrinogen Congenital Photosensitivity, blisters,

III synthase Erythropoietic atrophic scarring, hypertrichosis,
Porphyria erythrodontia, Port wine urine
5 Uroporphyrinogen Porphyria cutanea Photosensitivity

decarboxylase Tarda
6 Coproporphyrinogen Hereditary Both photosensitivity and

Oxidase Coproporphyria neuropsychiatric manifestations
7 Protoporphyrinogen Variegate Porphyria Both photosensitivity and

Oxidase neuropsychiatric manifestations
8 Ferrochelatase Erythropoietic Photosensitivity

protoporphyria
Classification of Porphyrias
WITH
NEUROPSYCHIATRIC WITH PHOTOSENSITIVITY BOTH
MANIFESTATIONS
1. ALA DEHYDRATASE 1.CONGENITAL

ERYTHROPOIETIC PORPHYRIA 1. VARIEGATE PORPHYRIA
DEFICIENT PORPHYRIA
2.PORPHYRIA CUTANEA TARDA 2. HEREDITARY
2. ACUTE INTERMITTENT
COPROPORPHYRIA
PORPHYRIA 3.HEREDITARY ERYTHROPOIETIC
116 >>>
MCQS
1. A 35-year-old man has a history of 3.The most common porphyria is
intermittent abdominal pain and episodes
of confusion and psychiatric problems. High A. PCT
amounts of δ-aminolevulinate and por- B. ADP
phobilinogen is also detected in his urine C. HCP
analysis. The patient also has a mutation in D. AIP
4.Which of the following porphyrias is

the gene for uroporphyrinogen I synthase
(porphobilinogen deaminase). The probable
diagnosis of the patient is: autosomal recessive
A. X-linked sideroblastic anemia A. PCT

B. Acute intermittent porphyria B. ALA synthase defect
C. Congenital erythropoietic porphyria C. CEP
D. Porphyria cutanea tarda D. AIP
2. All of the following are associated with

photosensitivity except
Autosomal Recessive
A. HEP Porphyrias
B. EPP
C. PCT • ADP
D. AIP • CEP
• HEP
5.A paint shop keeper presents with

anemia. Which of the following enzymes’
inhibiton causes anemia?
A. ALA synthase
B. Uroporphyrinogen III synthase
C. Uroporphyrinogen I synthase
D. Ferrochelatase
ALA DEHYDRATASE THE MOST SENSI-
TIVE MARKER FOR
1.B; 2.D; 3.A; 4.C; 5.D
COPROPORPHYRINOGEN LEAD POISONING

LEAD POSIONING IS ZINC PRO-
OXIDASE
TOPORPHYRIN
LEVELS
FEEROCHELATASE
117
Lipid Chemistry
Classification
What are lipids?
• Heterogenous group of substances which • Simple lipids are esters containing fatty
are non polar acids and alcohol
• They are of three types – Simple lipids, Com-
plex lipids and precursor or derived lipids
STRUCTURE OF TRIACYLGLYCEROL
• Heterogenous group of substances which • If the alcohol is a low molecular weight

are non polar alcohol like glycerol it is fat e.g., Triacyl
• They are of three types – Simple lipids, glycerol
Complex lipids and precursor or derived • If the alcohol is a high molecular weight
lipids alcohol like sphingosine, it is wax eg.,
• Simple lipids are esters containing fatty ceramide
acids and alcohol
119
What is sphingosine?
• Sphingosine is an amino alcohol i.e., it • Serine reacts with palmitoyl CoA to form
has an amino group and a hydroxyl group sphingosine
• The precursor of sphingosine is an amino • Sphingosine reacts with fatty acids to
acid with hydroxyl group – serine. form ceramide. Fatty acids react with
amino group of sphingosine to form an
amide linkage, hence the name ceramide.
SPHINGOSINE
FATTY
ACID
SERINE PALMITOYL CoA SPHINGOSINE
R-COOH
AMIDE
LINKAGE
R-COOH

120 >>>
CERAMIDE
LIPIDS
SIMPLE COMPLEX PRECURSOR/ DERIVED
ESTERS
FATTY ACID ALCOHOL
LIPIDS
FAT WAX
GLYCEROL SPHINGOSINE
TRIACYLGLYCEROL CERAMIDE
What are complex lipids?

• Complex lipids are esters containing » Glycolipids
some other group in addition to fatty acid » Aminolipids
and alcohol » Sulpholipids
• Depending upon the additional group • Phospholipids are complex lipids which
that is present, complex lipids are classi- have phosphate groups in addition to
fied into four types: fatty acids and alcohol
» Phospholipids
121
• Glycolipid are complex lipids which have • Aminolipids and sulpholipids are just
carbohydrate groups in addition to fatty derivatives of phospholipids and glycolip-
acids and alcohol ids eg., sulphogalactosyl ceramide is an
important component of myelin
LIPIDS
ESTERS
???? FATTY ACID ALCOHOL

PHOSPHOLIPID GLYCOLIPID AMINOLIPID SULPHOLIPID
Sulphogalactos
ylceramide
What are Phospholipids?
>> Phospholipids are complex lipids which » Choline, it. Is called as phosphatidyl
have phosphate groups in addition to fatty choline or lecithin eg., Dipalmitoyl phos-
acids and alcohol phatidyl choline is the major component
>> Depending upon the alcohol that is of surfactant
present, phospholipids are classified as » Serine, it is called as phosphatidyl serine
• Glycerophospholipid » Inositol, it is called as phosphatidyl ino-
• Sphingophospholipid sitol
>>Glycerophospholipid: >> Sphingophospholipid:
• They are esters containing phosphate • They are esters containing phosphate
group in addition to glycerol and fatty group in addition to fatty acid and
acid. They are derivatives of phosphati- sphingosine. Fatty acid and sphingosine
date are called as ceramide. Hence sphin-
gophospholipids are derivatives of
• In phosphatidate, the first two hydroxyl
ceramide
groups of glycerol are attached to 2
fatty acids, and the third hydroxyl group • If ceramide is attached to phosphate
is attached to phosphate group. If the and choline, that is sphingomyelin.
phosphatidate group is attached to
122 >>>
PHOSPHOLIPIDS
ESTERS
PHOSPHATE FATTY ACID ALCOHOL
GLYCEROPHOSPHOLIPIDS SPHINGOPHOSPHOLIPIDS
Glycerophospholipids
ESTERS
PHOSPHATE FATTY ACID GLYCEROL
Phosphatidate
CHOLINE lecithin
OC – R1
SERINE PS
OC – R2 INOSITOL PI
ETHANOLAMINE CEPHALIN
O
I DIPHOSPHATIDYL
P–O CARDIOLIPIN
GLYCEROL
I
O ETHER PLASMALOGENS
What are precursor or

derived lipids?
• Any other non-polar substance, which rived from them are called as precursor
has not been included in the simple lipid or derived lipids
or complex lipid category, but which can • Example - Fatty acid, cholesterol, bile
act as precursors of them or can be de- acids, steroid hormones
123
LIPIDS
Fatty acid, Cholesterol

and Bile acid
ONE LINERS
• Sphingosine and fatty acid is called as Ceramide
• The aminoacid precursor of sphingosine is Serine
• hosphatidyl Choline is Lecithin
• Phosphatidyl ethanolamine is Cephaline
• Diphosphatidyl glycerol is Cardiolipin
MCQS
1.Which of the following is not a lipid 4.Which of the following is a derived lipid
A. Glycerol A. Triacyl glycerol
B. Palmitic acid B. Fatty acid
C. Triacylglycerol C. Sphingomyelin
D. Cholesterol D. Cholesterol
2. All of the following are components of 5. All the following are components of
triacyl glycerol except sphingomyelin except
A. Glycerol A. Sphingosine
B. Palmitic acid B. Fatty acid
C. Sphingosine C. Choline
D. Stearic acid D. Ethanolamine
3.Which of the following is a wax

A. Triacyl glycerol
B. Ceramide
C. Sphingomyelin
D. Lecithin
124 >>>
INTEGRATED CASE BASED MCQ
6. A premature child presents with Infant

Respiratory Distress Syndrome. The neona-
tologist says the neonate is yet to produce
lecithin adequately. All of the following are
components of lecithin except
A. Glycerol
B. Choline
C. Phosphatidate
D. Ceramide
IMAGE BASED MCQS
7. Viper venom has Phospho- 8. Products of the enzyme X

1.A; 2.C; 3.B; 4.D; 5.D 6.D; 7.B; 8.B.
lipase A2. Phospholipase A2 are

activity is shown as in
the image A. Lysophospholipid and fatty
acid
A. A B. Inositol triphosphate and Dia-
B. B cyl Glycerol
C. X C. Phosphatidic acid and a ni-
D. Y trogenous base
D. Inositol triphosphate and
phosphatidic acid
125
Glycolipids
Glycosphingolipids
• They are complex lipids which have • If ceramide is attached to an oligosaccha-
carbohydrate group in addition to fatty ride, it is called as globoside eg., Lactosyl
acid and alcohol – sphingosine. Fatty acid ceramide
and sphingosine are called as ceramide. • If ceramide is attached to an oligosaccha-
Hence glycolipids are derivatives of cer- ride with NANA, it is called as ganglioside
amide eg., Cer – Glu- Gal- NANA is GM3 ganglio-
• If ceramide is attached to monosac- side. As complexity increases, the num-

charide, it is called as cerebroside. Eg., ber decreases.
Glucosyl Ceramide
Glycolipids
ESTERS
CARBOHYDRATE FATTY ACID SPHINGOSINE
CERAMIDE
Cer - GLU/ GAL CEREBROSIDE
Cer - GLU/ GAL GLOBOSIDE
Cer - Glu - Gal - NANA GM3 GANGLIOSIDE
N – Acetyl Gal
GM2 GANGLIOSIDE
NH2
Gal GM1 GANGLIOSIDE
126 >>>
Introduction to Glycosphingolipidosis
• The metabolism of GM1 ganglioside be- • Globoside is metabolized by the removal
gins with beta galactosidase removing of galactose by alpha galatosidase to
the galactose from GM1 ganglioside and form cerebroside. Hence alpha galac-
converting it to GM2 ganglioside. Hence tosidase defect causes Fabry’s disease
defect of beta galactosidase causes characterized by the accumulation of
GM1 gangliosidoses globosides
• GM2 ganglioside metabolism is initiated • Cerebroside is metabolized by the re-
by removal of N-Acetyl Galactosamine moval of glucose by beta glucosidase.
by Galactosaminidase or Hexosamini- Beta glucosidase defect causes Gauch-
dase to form GM3 ganglioside. Hence er’s disease
Hexosaminidase defect causes GM2 • Ceramide is metabolized by ceramidase
gangliosidoses to form sphingosine and fatty acid.
• GM3 ganglioside is metabolized by the Defect of ceramidase causes Farber’s
removal of N Acetyl Neuraminic acid by disease, which is a granulomatous
Neuraminidase to form globoside. De- disorder and is characterized by pain-
fect of Neuraminidase causes sialidoses ful subcutaneous nodules and chronic
kidney disease
Glycosphingolipidosis
S.No Enzyme Defect Disorder
127
S.No Enzyme Defect Disorder
1 Beta Galactosidase GM1 Gangliosidosis
2 Alpha Galactosidase Fabry’s disease
3 HexosaminidaseA Tay Sach’s disease
4 HexosaminidaseA and B Sandhoff’s disease
5 Neuraminidase Sialidosis
6 Beta Glucosidase Gaucher’s disease
7 Ceramidase Farber’s disease

S.No Enzyme Defect Disorder CLINICAL FEATURES
1 Beta GM1
Galactosidase Gangliosidosis
2 Alpha Fabry’s disease

Galactosidase
3 HexosaminidaseA Tay Sach’s

disease
4 HexosaminidaseA Sandhoff’s
and B disease
5 Neuraminidase Sialidosis
6 Beta Gaucher’s
Glucosidase disease
7 Ceramidase Farber’s
disease
128 >>>
S.No Enzyme Defect Disorder CLINICAL FEATURES
1 Beta GM1 Neurological manifestations and Systemic

Galactosidase Gangliosidosis manifestations
2 Alpha Fabry’s disease Reddish purple spots, CKD, early MI

Galactosidase
3 HexosaminidaseA Tay Sach’s Neurological manifestations, Cherry red

disease spots, No Organomegaly
4 HexosaminidaseA Sandhoff’s Neurological manifestations, Cherry

and B disease red spots, No Organomegaly – GM2
ganglioside and globoside accumulation
5 Neuraminidase Sialidosis Generalised swelling, Neurological and

organomegaly
6 Beta Gaucher’s Anemia, thrombocytopenia, HSM, Tissue

Glucosidase disease paper cells
7 Ceramidase Farber’s Granulomatous disorder

disease
Facts about Glycosphingolipidosis

• No MR – Gaucher’s & Fabry’s • GM2 Gangliosidoses - Tay Sach’s and
• No Cherry Red spot – Gaucher’s, Fabry’s Sandhoff’s
and Krabbe’s • Sandhoff’s – accumulation of both glo-
• No HSM – GM2 gangliosidosis, Krabbe’s boside and ganglioside
disease
Difference between Taysach’s &

Sandhoff
S.No PROPERTY TAYSACH’s SANDHOFF
1 Gene defect Hex A Hex B
2 Subunit Alpha Beta
3 Enzyme defective Hex A Hex A and B
4 Lipid accumulation GM 2 ganglioside GM2 ganglioside and globoside
129
MCQS
1.A child presented with refractory ane-
mia. Below is provided the smear of bone
marrow biopsy of the child. The probable
enzyme deficiency is?
A. Beta Glucocerebrosidase
B. 1,4 Alpha glucosidase
C. Hexosaminidase A
D. Hexosaminidase B
2. A child presented with difficulty in

vision on examination cherry red spots were • Cherry red spot is absent in KGF
seen on macula. There was no organomega- • Krabbe’s disease
• Gaucher’s disease
ly. Identify the disease.
• Fabry’s disease
A. Gaucher disease
B. Hunter disease
CHERRY RED SPOT
C. Tay-sach disease
D. Niemann pick disease
3. A young boy is having difficulty in

• Opacification of remaining parts
of macula
• Fovea retains the transparency
breathing while running, and is unable to • Lipid storage disorders
rise from squatting position. His fundus • Fovea lacks ganglion cells
examination image is provided below. The • Central retinal artery occlusion
probable condition is? • Fovea receives blood supply from
choroid
A young boy is having difficulty in breathing
while running, and is unable to rise from
squatting position. His fundus examination
image is provided below. The probable
condition is?
A. Tay Sachs Disease

B. Gaucher’s disease
C. Fabry’s disease
D. Krabbe’s disease
130 >>>
4. A child presents with anemia, throm- 5. For the first few months, an eight-
bocytopenia. On examination, hepatos- month-old child’s growth and develop-
plenomegaly is observed. The child also ment were normal, then symptoms such
complains of bony pain. Light microscopy as deafness, blindness, atrophied muscle,
reveals crumpled tissue paper appear- inability to swallow, and convulsions began
ance. The enzyme defect is ? to appear. During the fundus inspection, a
cherry red macula was also seen in both
A. Alpha glucosidase eyes. Suspecting sphingolipidosis, the physi-
B. Beta glucosidase cian tested for globosides and gangliosides
C. Alpha galactosidase levels and found that both were increased.
D. Beta galactosidase Based on this finding, what is the most accu-
1.A; 2.C; 3.A; 4.B; 5.A;

rate diagnosis for this patient?
GLUCOSYL
CERAMIDE
A. Sandhoff’s disease
MACROPHAGES B. Tay Sach’s Disease
C. Gaucher’s disease
TISSUE PAPER CELLS D. Fabry’s disease
Classification of lipids
based on solubility
Cholesterol structure
• Cyclo Pentano Perhydro Phenantherene

ring
• Hydroxyl group at 3rd position
• Methyl groups at 10th and 13th position
• Side chain at position 17
NO POLAR CHOLESTEROL
GROUP NEUTRAL LIPID ESTER
NON TAG
LIPIDS
POLAR
POLAR AMPHIPATHIC CHOLESTEROL

LIPID PHOSPHOLIPID
GROUP
131
Fatty Acids
CH3–CH2–CH2–CH2–CH2-CH2–CH2–CH2–CH2–CH2-CH3–CH2–CH2–CH2–CH2–CH2–CH2-COOH
ω1 ω2 ω3 △4 △3 △2 △1
• If you start numbering from the carboxyl end, carboxyl carbon is △1

• If you start numbering from the methyl end, the methyl carbon is ω1
• ωn = N - △n
• ω9 fatty acid is a fatty acid in which he first double bond is next to the 9th carbon atom
when you move from the methyl end
• ω6 fatty acid is a fatty acid in which he first double bond is next to the 6th carbon
atom when you move from the methyl end
CH3–CH2–CH2–CH2–CH2-CH2–CH2–CH2–CH2–CH2-CH3–CH2–CH2–CH2–CH2–CH2–CH2-COOH
β ⍺
Nutritionally essential Fatty Acids
• Linoleic acid (18: ω6) and alpha linolen- tem, only ω9 fatty acids can be synthe-
ic acid (18: ω3) are nutritionally essen- sised
tial • So ω6 and ω3 fatty acids have to be
• Human enzymes can not desaturate be- supplied in diet
yond Δ9. Hence for a fatty acid with 18 • Arachidonic acid is a semiessential fatty
carbon atom, which is most commonly acid.
used by human desaturase enzyme sys-
132 >>>
MCQS
1. All of the following are true about cho- 4.Arachidonic acid is a fatty acid of
lesterol except which type?
A. Hydroxyl group at 3rd position A. 20:ω6

B. Methyl groups at 10th and 13th position B. 20:ω3
C. Side chain at 17th position C. 18:ω6
D. Has benzene ring D. 22:ω3
2. Which of the following is a neutral 5. Which of the following is nutritionally

lipid? essential?
A. Phospholipid A. Arachidonic acid

B. Monoacyl glycerol B. Linoleic acid
C. Diacyl glycerol C. Cervonic acid
D. Cholesterol ester D. Oleic acid
3.Which of the following is not amphipa-

thic?
A. Triacylglycerol
B. Phosphoglycerol
C. Phospholipid
D. Gycolipid
1.D; 2.D; 3.A; 4.A; 5.B
133
Lipid
Metabolism
Fatty Acid Synthesis

Introduction
• Acetyl CoA is the building block of fatty • Low energy status:
acids. Acetyl CoA is a central metabolite » TCA cycle and is oxidized as Carbon
and it can have various fates. Fates of dioxide
acetyl CoA are dependent on the ener- » If oxaloacetate is not available, it
gy status of the cell forms ketone bodies
• FATES OF ACETYL COA • Acetyl CoA carboxylase converts acetyl
• High energy status: CoA to malonyl CoA and thereby com-
» Fatty acid synthesis mits acetyl CoA to fatty acid synthesis.
» Cholesterol Synthesis Hence acetyl CoA carboxylase is the rate
limiting enzyme of fatty acid synthesis
RATE LIMITING ENZYMES OF LIPID METABOLISM
135
1 Fatty Acid Synthesis Acetyl CoA carboxylase
2 Fatty Acid Oxidation Carnitine Acyl Transferase I
3 Bile Acid Synthesis 7 Alpha Hydroxylase
4 Ketone body synthesis HMG CoA Lyase
5 Cholesterol Synthesis HMG CoA reductase
Facts about Fatty Acid Synthesis

• Fatty acids get synthesised in cytoplasm mitochondria to cytoplasm. In cyto-

• Acetyl CoA is the building block plasm, ATP citrate lyase cleaves cit-
• Sources of acetyl CoA: rate to form acetyl CoA and Oxaloace-
tate. Acetyl CoA enters into fatty acid
» PDH in mitochondria
synthesis and oxaloacetate is convert-
» Fatty acid oxidation in mitochondria ed by malate dehydrogenase to form
» Aminoacid oxidation in mitochondria malate. Malic enzyme converts malate
• Prerequisite for fatty acid synthesis: to form pyruvate. This forms NADPH,
» Transport of acetyl CoA from mito- which acts as the source of NADPH
chondria to cytoplasm for fatty acid synthesis.
» Acetyl CoA reacts with oxaloacetate • Acetyl CoA carboxylase is the rate limit-
to form citrate and the tricarboxylate ing enzyme
transporter transorts citrate from • Fatty acid synthase complex is the other
enzyme complex involved
Facts about Fatty Acid Synthase

Complex
• It is present in Cytoplasm » 7 enzymes
• It has two units • Simultaneously two fatty acids get syn-
• Each unit has the following features thesised. One half of each unit works to-
» A cysteine end gether to synthesise a fatty acid. Hence,
subunit division is not the same as the
» A 4-phosphopatothiene end
functional division
» 8 subunits
136 >>>
Steps of Fatty Acid Synthesis

• Acetyl CoA carboxylase converts acetyl • This continues until a 16 carbon atom
CoA to malonyl CoA containing fatty acid or palmitic acid is
• Acetyl transacylase transfers acetyl group synthesized in the panothiene end
to the cysteine end • Thioioesterase makes the fatty acid free
• Malonyl transacylase transfers malonyl
group to phosphopantothiene end
• Ketoacyl synthase transfers the acetyl
group from cys end to malonyl group. This
results in decarboxylation and the ketoa-
cyl group is formed in the pantothiene end
• Ketoacyl reductase reduces the ketoacyl
group to form hydrooxyacyl group
• Hydratase removes water and introduces
a double bond to form enoyl acid
• Enoyl reductase reduces an unsaturated
acid to form saturated acid
• By this time acetyl coA carboxylase would
have made a malonyl group ready, which
will be transferred by malonyl transacylase
to the pantothiene end.
• This shifts the fatty acid that is synthesized
in the previous cycle to the cys end.
SUBUNITS OF FATTY ACID SYNTHASE COMPLEX

» Acetyl Transacylase » Hydratase
» Malonyl Transacylase » Enoyl reductase
» Ketoacyl Synthase » Thioestaerase
» Ketoacyl reductase » Acyl Carrier protein
REGULATION OF ACETYL COA CARBOXYLASE
• Stimulated by high » Citrate » NAD

energy » Insulin » FAD
» ATP » Acyl CoA
» NADH • Inhibited by low energy » Glucagon
» FADH2 » ADP
137
MCQS
1. The precursor of fatty acids is 5. All are true about Fatty acid synthase,
EXCEPT :
A. Propionyl CoA
B. Malonyl CoA A. It is Dimer
C. Acetyl CoA B. Acetyl CoA carboxylase is the first
D. Methyl Malonyl CoA enzyme of the complex
2. All the following are fates of acetyl

C. Pantothenic acid is a component of the
enzyme
CoA except D. It has got 7 enzmes, in each unit
A. CO2 6. The most synthesized fatty acid by

B. Ketone body fatty acid synthase complex is
C. Cholesterol
D. Glucose A. Linoleic acid
3.The rate limiting enzyme of fatty acid

B. Palmitic acid
C. Arachidonic acid
synthesis is D. Stearic acid
A. HMG CoA reductase 7. Lipogenesis is stimulated by all,

B. HMG CoA Lyase EXCEPT :
C. Acetyl CoA carboxylase
D. 7 Alpha hydroxylase A. High fatty diet
4. All the following are requirements of

B. High Glucose diet
C. High Fructose based diet
Acetyl CoA carboxylase except D. Insulin
A. Bicarbonate
B. Biotin
1.C; 2.D; 3.C; 4.D; 5.B; 6.B; 7.A
C. ATP
D. NADH
138 >>>

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BIOCHEMISTRY

Carbohydrate Enzyme Kinetics and Enzyme

Enzymes 86 Fatty acids 132

Introduction 87 Lipid metabolism 134

Clasification of Enzymes 87 Fatty acid synthesis 135

Pathways and Suborganelles

TCA Cycle >

Glycogen metabolism >

HMP SHUNT >

Fatty Acid Synthesis >

Fatty Acid Oxidation >

Ketone body synthesis >

Cholesterol synthesis >

Bile acid Synthesis >

Urea Cycle >

Heme synthesis >

Glycolysis > Cytoplasm

PDH > Mitochondria

TCA Cycle > Mitochondria

Glycogen metabolism > Cytoplasm

Gluconeogenesis > Mitochondria,

HMP SHUNT > Cytoplasm

Fatty Acid Synthesis > Cytoplasm

Fatty Acid Oxidation > Mitochondria and

Cholesterol synthesis > Cytoplasm and ER

Ketone body synthesis > Mitochondria

Bile acid Synthesis > ER

Urea Cycle > Mitochondria and

Heme synthesis > Mitochondria and

Other points to remember: Suborganelles

• Ether lipid synthesis

S.No Organelle Marker enzyme

2 Endoplasmic reticulum >

3 Golgi complex >

S.No Organelle Marker enzyme

1 Nucleus > DNA polymerase/

2 Endoplasmic reticulum > Glucose 6

3 Golgi complex > Galactosyl

4 Mitochondria Outer > Monoamine Oxidase

Inner > ATP synthase

5 Lysosome > Cathepsin

6 Cytoplasm > Lactate

7 Peroxisome > Catalase

Other points to remember: Marker enzyme

• Replication and Transcription occur in • Marker enzykme of Outer Membrane

• Golgi complex is related to glycopro- drogenase and Complex V or ATP syn-

tein synthesis. The enzyme involved thase

in Glycoprotein synthesis is galactosyl • Hydrogen peroxide is generated Per-

Where are proteins synthesised?

2. The marker enzyme of microsomes is

A. Galactosyl transferase 6. A 39 year old man came to the emer-

3. Cytoplasmic proteins are synthe-

4.Secretory proteins are synthesized in?

C. Rough endoplasmic reticulum

5. A child presents with coarse features,

types – Glycoproteins, Proteoglycans

ly complex carbohydrates are of three sugar units

Comparison of glucose & fructose

S.No Disaccharide Sugar Linkage

1 Maltose Glu + Glu α (1,4)

2 Isomaltose Glu + Glu α (1,6)

3 Trehalose Glu + Glu α (1,1)

4 Sucrose Glu + Fru α (1,2)

5 Lactose Gal + Glu β (1,4)