Enz ymes

Universities
Enz ymes- Specific Learning Objectives
(1)define enzyme, active site, substrate.
(2)classify enzymes with suitable examples using IUB
classification.
(3)explain the role of coenzymes in enzyme action with
suitable examples
(4)explain the functions of coenzymes with suitable
examples
(5)enumerate various cofactors required in metabolic
reactions
(6)Explain the mechanism of enzyme action
(7)Explain the Regulation of Enzyme Activity
(8)Enzyme Inhibition
(9)explain isoenzymes with their salient features
 ENZYMES
Enzymes are biocatalysts that
increase the velocity of a
chemical reaction and are
neither consumed nor changed
during the reaction.
 Enzymes as catalysts
• Catalyst - Speeds up
chemical reactions in
living organisms by
decreasing the energy Without a catalyst

needed to start the Energy

reaction (activation
energy)
With a catalyst

Time
 Definitions

• Substrate- monomers that bind to

the active site of an enzyme
• Active site- area on enzyme where
substrate binds
• Product- what the enzyme produces
 Characteristics of Enzymes
Proteinous nature of enzymes:-
• All enzymes are protein in nature except ribo-
zymes ( catalytic RNA).
• They are heat labile.
• They are water – soluble.
• Precipitated by protein precipitating reagents.
• Contain 16% weight as nitrogen
• Enzymes are composed of one or more polypeptide
chains (subunit).

• Monomeric :- composed of single polypeptide

chain.

• Oligomeric:- composed more than one subunits.Eg:-

Phosphofructokinase (PFK) & Creatine kinase- with two
subunit.
Glycogen phosphorylase & Lactate dehydrogenase (LDH)-
with four subunit.
• Multiple enzyme complex:- Exist in cluster or group.
e.g:-
Fatty acid synthase complex:- seven different
enzymes- acetyl CoA transacylase, malonylCoA
transacylase, β-ketoacyl synthase, β-ketoacyl
reductase, hydroxyacyl dehydratase, enoyl reductase
and thiolase

Pyruvate dehydrogenase:- three enzymes- Pyruvate

dehydrogenase, dihydrolipoyl transacetylase and
dihydrolipoyl dehydrogenase.
 CHAPTER 7

Active Site
The small cleft- like portion of an enzyme where the
substrate(s) binds and catalysis occurs is known as the
active site or active centre.

• Subst rate binding

• site Catalytic site

A diagrammatic representation of
an enzyme and its active site

Rafi M D: Textbook of Biochemistry (4 th

Edition) Universities
 Coenzymes
(Non-protein part or prosthetic group)

(Protein part of enzymes)

Coenzymes may divided into two groups.

Coenzymes Coenzymes of B- complex vitamins
 CHAPTER 7

Coenzymes which are not related to vitamins

 Metallo-enzymes
Enzymes which require certain metals for their activity
are called metallo-enzymes .Metal ions is cofacter.

Metal Enyzmes
Cu2+ CytochromeC oxidase, Cerruloplasmin, SOD

Fe2+ CytochromeC oxidase, Catalase, Peroxidase

Se Glutathione peroxidase
Mo Xanthine oxidase
Mn DNA polymerase,
Ni2+ Urease
Mg Hexokinase, Phosphofructokinase, Enolase
Zn Carbonic anhydrase, Alcohol dehydrogenase
 Specificity of enzymes
Enzymes are highly specific, interacting
with one or a few specific substrates and
catalyzing only one type of chemical
reaction.
• Absolute or substrate specificity:- the
enzyme acts only on one substrate. e.g.
a) Urease, which act only on urea.
b) Carbonic anhydrase, act on carbonic acid.
c) Lactose, which act on lactose
d) Sucrase,which act on sucrose.
e) Maltase, which act on maltose.
• Group specificity:-Enzyme catalyzes reaction
involving any molecules with the same functional
group.
• Bond specificity:- enzyme catalyzes the
formation or break up of only certain type of
bond. eg:

a) Amylase, act on 1-4 glycosidic bond on

starch, dextrin & glycogen.
b) Lipase ,hydrolyzes ester bonds in different
triglycerides.
• Stereo-specificity :- enzyme act only one type
of isomer.
a) L amino acid oxidase acts only on L amino
acids.
b) D amino acid oxidase acts only on D amino
acids.
c) α-Glycosidase acts only on α-glycosidic bonds ,
which are present in starch, dextrin and
glycogen.
d) β-Glycosidase acts only on β-glycosidic bonds.
 Nomenclature and Classification
Enzymes are classified into following categories
according to the reactions that they catalyze:
1. Oxidoreductase
2. Transferase
3. Hydrolase
4. Lyase
5. Isomerase
6. Ligase
 1-Oxidoreductases
• Oxidoreductases catalyze redox reactions
– Reductases
– Oxidases
AH2+B A+BH2

NADHperoxidase

NADH++H++H2O2 Glutathione reductaseNAD

+
+H2O
G-S-S-G+NADH++H+ 2G-SH+NAD+
 2-Transferases
• Transferases transfer a group from one molecule to
another
– Transaminases catalyze transfer of an amino group
– Kinases transfer a phosphate group

Ketoglutarate+aspartate Glutamate Oxaloacetate Glutamate+ Oxaloacetate

Transaminases (GOT)

ATP+D-fructose-6-phosphate Phosphofructokinase ADP+D-fructose-1-6-

bisphosphate
 3-Hydrolases
• Hydrolases cleave bonds by adding water
– Phosphatases
– Digestive enzymes- Peptidases, Amylase, Lipases
G-6-Pase
D-Glucose-6-phosphate+H2O D-glucose+Pi
Sucrase
Sucrose+H2O D-Glucose+D-Fructose
 4-Lyases
• Lyases catalyze cleavage and formation of bonds
such as C-C, C-N or C-S without involvement of
water.
– Decarboxylases
– Aldolases
Fructose-1-phosphate Aldolase B Dihydroxyacetone
phosphate+glyceraldehyde

Histidine Histidine decarboxylase Histamine+CO2

 5-Isomerases
• Isomerases catalyze isomerisation reactions
– Epimerases
– Mutases
– Isomerases
Phosphoglycerate mutase
3Phosphoglycerate 2Phosphoglycerate

UDP-hexose-4-epimerase
UDP-glucose UDP-galactose

Phosphohexose isomerase
D-glucose-6-phosphate D-fructose-6-phosphate
 6-Ligase
• Ligases catalyze a reaction in which formation of C-
C, C-S, C-O, or C-N bond coupled to hydrolysis of
ATP. Pyruvate carboxylase
Pyruvate + CO2+ATP Oxaloacetate+ADP+Pi
Carbamoyl phosphate synthase I
Ammonia+CO2+2ATP Carbamoyl phosphate
+2ADP+Pi
 Nomenclature of Enzymes
• In most cases, enzyme names end in –ase
• The name of an enzyme identifies the reacting substance. For example,
– Urea: remove -a, replace with -ase = urease
– Lactose: remove -ose, replace with -ase = lactase
– sucrase catalyzes the hydrolysis of sucrose
• The name also describes the function of the enzyme For example,
- oxidases catalyze oxidation reactions
- Hydrolase
• Some enzymes are named for the substrate and the function. For
example,
– Lactate dehydrogenase
– Pyruvate decarboxylase
– Alcohol dehydrogenase
• Some names are historical - no direct relationship to substrate or
reaction type
– Catalase
– Pepsin
– Chymotrypsin
–
 Mechanism of Enzyme Action
• Enzyme binds to the substrate and form an
enzyme-substrate(ES) complex.

Substrate is bound
through multiple
non-covalent
bonds at the
active site.
 Active site
• The region of the enzyme combine with the
substrate .
• Active sites characteristics include:
– Pockets or clefts in the surface of the enzyme.
– R groups at active site are called catalytic groups.
– The active site contains amino acid side chains that create a
three-dimensional surface complimentary to the shape of
the substrate.
– The enzyme attracts and holds the enzyme using weak non-
covalent interactions (H-bonding, hydrophobic
interactions).
– Conformation of the active site determines the specificity of
the enzyme.
– Products are released when the reaction is complete (they
no longer fit well in the active site)
Fischer Template Model (Lock and Key Model)
• This is a rigid model of catalytic site, proposed by Emil Fischer in
1894.
• In the lock-and-key model, the enzyme is assumed to be the lock
and the substrate the key
– The active site has a rigid shape
– Only substrates with the matching shape can fit
– The enzyme and substrate are made to fit exactly
– This model fails to explain change in enzyme structure in the presence
of allosteric modulators.
 Lock and Key Model
Two substrates

Enzyme

Active site of the enzyme

 Lock and Key Model
The substrates fit like a key in a lock

Enzyme

The active site is like a lock

 Lock and Key Model
The activation energy for these substrates
to bind together has been lowered by the
enzyme.

Chemical reaction!!!

Enzyme
 Basic Enzyme Diagram
The substrates have
reacted and changed
into the product

Enzyme is unchanged

Active site
 Induced Fit Enzyme Model
• Koshland proposed an induced-fit model in 1963.
-The active site is flexible, not rigid
- the shapes of the enzyme, active site, and substrate adjust
to maximize the fit, which improves catalysis
- there is a greater range of substrate specificity
• This model is more consistent with a wider range of enzymes
How does the enzyme promote a faster
chemical reaction?
– As the substrate interacts with the enzyme, its shape
changes and this new shape is less energetically stable
– This transition state has features of both substrate and
product and falls apart to yield product, which
dissociates from the enzyme
 Possible Types of Transition State Changes

The enzyme might put “stress” on a bond facilitating bond

breakage
 FACTORS AFFECTING ENZYME
ACTIVITY
Major factor responsible for the enzyme activity are:
• Temperature
• pH
• Time
• Concentration of substrate
• Concentration of enzyme
• Concentration of products
 Temperature and Enzyme Activity
• Enzymes are most active at an optimum temperature (usually
37°C in humans)
• They show little activity at low temperatures
• Activity is lost at high temperatures as denaturation occurs

Bell shaped
Majority of curve
enzymes
completely
lost their
catalytic
activity at
about >70⁰C.
 pH and Enzyme Activity
• Enzymes are most active at optimum pH
• Amino acids with acidic or basic side-chains have the proper
charges when the pH is optimum
• Activity is lost at low or high pH as tertiary structure is disrupted

•At alkaline pH this group is

deprotonated, and the rate of
reaction therefore declines.
•Extremes acidic pH can also lead to
denaturation of the enzyme.
•A pH change in charge state of
substrate impairs its binding to active
site.
•Clinical conditions such as alkalosis
and acidosis may alter enzyme
activity.
 Optimum pH for Selected Enzymes
• Most enzymes of the body have an optimum pH of about 7.4
• However, in certain organs, enzymes operate at lower and
higher optimum pH values
 Enzyme Concentration and Reaction Rate
• Reaction rate is directly proportional to the concentration of enzyme
• The rate of reaction increases as enzyme concentration increases (at
constant substrate concentration)
• At higher enzyme concentrations, more enzymes are available to catalyze
the reaction
• There is a linear relationship between reaction rate and enzyme
concentration (at constant substrate concentration)
 Substrate Concentration and Reaction Rate

• The rate of reaction increases as substrate concentration increases

(at constant enzyme concentration)
• Maximum activity occurs when the enzyme is saturated (when all
enzymes are binding substrate)
 Concentration of products
• Accumulation of products of reaction causes
inhibition of enzyme activity.

SUBSTRATE PRODUCT

SUBSTRATE × PRODUCT
 ENZYME KINETIC
• At a fixed enzyme concentration, reaction velocity (V) of an
enzyme-catalyzed reaction increases with rise in substrate
concentration (S) until a maximal velocity (Vmax) is attained.
• Most enzymes show hyperbolic curve when reaction velocity is
plotted against substrate concentration.
A- at low substrate Hyperbolic curve
C
concentration, reaction
B
velocity increase linearly with
S.
B- at high substrate
concentration, velocity is
A
increase but not directly
proportional to S.
C- Vmax is attained ,rise in S
does not further increase the
reaction.
MICHAELIS-MENTEN EQUATION
• Michaelis and Menten proposed a model 1913.
• To explain kinetic features of enzyme-catalyzed reactions.
• In this model, the enzyme reversibly combines with its substrate
to form an ES complex that subsequently breaks down to product,
regenerating the free enzyme.

• Michaelis-Menten equation

Where, Vo is initial reaction velocity

Vmax is maximal reaction velocity
Km is Michaelis-Menten constant
S is substrate concentration
Characteristics of Km:

• Km , The Michaelis constant is equal to the substrate

concentration at which reaction rate is half of its
maximal velocity.
• Km does not vary with the concentration of enzyme.
• Its measure of the affinity of the enzyme for its
substrate because a low Km indicates strong bonding
with substrate and high Km indicates weak binding.
 Lineweaver –Burk equation
• A more accurate method of determining
Vmax and Km.

This plot is useful to

determine the mechanism
of action of enzyme
inhibitors

Lineweaver –Burk Plot (Double reciprocal plot)

 Enzyme Inhibition
• It’s a phenomenon of decease in reaction
velocity is called enzyme inhibition.
• Chemicals can bind to enzymes and eliminate or
drastically reduce catalytic activity
• Inhibitors (I) are molecules that cause a loss of
enzyme activity
• They prevent substrates from fitting into the
active site of the enzyme:

E + S  ES  E + P
E + I  EI  no P formed
Classify enzyme inhibitors on the basis of reversibility

– Reversible Inhibitors:- inhibitors binds to the enzyme

through non-covalent bonds and activity of enzyme is restored
fully when inhibitor removed.
– Different type of reversible inhibitors are
– Competitive inhibitors often structurally resemble the
substrate and bind at the normal active site
– Non-competitive inhibitors usually bind at someplace other
than the active site
– Uncompetitive inhibitors only binds with ES complex.
– Irreversible inhibitors:- bind tightly to the enzyme
through covalent bonds and thereby prevent formation of the
E-S complex
 Competitive Inhibitors
• Reversible, competitive enzyme inhibitors are also
called structural analogs
– Molecules that resemble the structure and charge
distribution of a natural substance for an enzyme
– Resemblance permits the inhibitor to occupy the
enzyme active site
– Once inhibitor is at the active site, no reaction can
occur and the enzyme activity is inhibited
• Inhibition is competitive because the inhibitor and the
substrate compete for binding to the active site
– Degree of inhibition depends on the relative
concentrations of enzyme and inhibitor
19.10 Inhibition of Enzyme Activity
• The relative amounts of ES and EI complexes
depend upon the relative concentrations of the
substrate and the inhibitor
• If the inhibitor concentration is higher, more EI
complex will be formed resulting decreased
formation of the product.
• If substrate concentration is higher, more Es
complex will be formed, inhibition will be smaller
degree.
• Competitive inhibitor does not affect Vmax which
can be attained even in the presence of the
inhibitors
• But more substrate will be required to reach the
Vmax in the presence of the inhibitor.
• The inhibitors that rise the Km.
Some competitive inhibitors used as drugs

• Amethopterin and aminopterin

• Allopurinol
• Physostigmine and neostigmine
• Mevastatin and lovastatin
 • Amethopterin and aminopterin:- Structural
analogues of folic acid.
• Inhibitiors of dihydrofolate reductase
• Tetrahydrofolate is required in synthesis of
purine and thymine

Therefore amethopterin
and aminopterin used in
cancer to suppress cell
division
 Physostigmine and neostigmine

• Structural analogue of acetylcholine

• Inhibitors of acetyl cholinesterase
• Decrease the breakdown of acetylcholine
• Its used in treatment of myasthenia gravis
• Its an auto-immune disorder in which
acetylcholine receptors are decreased.
 Allopurinol
• Structural analogue of hypoxanthine.
• Inhibitor of xanthine oxidase.
• Gout can be treated by allopurinol drug therapy.
• Allopurinol is oxidized by xanthine oxidase to
alloxanthine, a potent inhibitor of xanthine oxidase.
• Less urate is produced.
 Mevastatin and lovastatin

• Structural analogue of HMG CoA

• Inhibitors of HMG CoA reductase
• Key enzyme of cholesterol synthesis.
• Decrease the synthesis of cholesterol.
• Therefore Mevastatin and lovastatin are used
as hypo-cholesterolaemic drugs
• Malonate is a competitive inhibitor of succinate
dehydrogenase
- it has a structure that is similar to succinate
- inhibition can be reversed by adding succinate
 Noncompetitive Inhibitors
• Reversible, noncompetitive enzyme inhibitors
bind to R groups of amino acids or to the metal
ion cofactors
– This binding is weak
– Enzyme activity is restored when the inhibitor
dissociates from the enzyme-inhibitor complex
– These inhibitors:
• Do not bind to the active site
• Do modify the shape of the active site once bound
elsewhere in the structure
 - it binds to an other site
rather than to the active site

- it distorts the shape of the

enzyme, which alters the shape
of the active site and prevents
the binding of the substrate
• This means that the non- competitive
inhibitors lower the Vmax but do not affect
the Km.
• Examples:- Ethanol, narcotic drugs are non-
competitive inhibitors of acid phosphatase.
• Trypsin inhibitors occurs in soybean and raw
egg white.
 Uncompetitive Inhibitors
• Uncompetitive Inhibitors can binds only to ES
complex
• It does not have affinity for free enzyme.
• This form of inhibitor is rare with single
substrate but more common with multiple
substrate reaction.
• Placental alkaline phosphotase is inhibited by
phenylalanine
•Uncompetitive inhibitors decreases both
Vmax and Km.
 Irreversible Inhibitors
• Irreversible inhibitors binds with enzyme very tightly
covalently and forms a stable complex.

– Binding of the inhibitor to one of the R groups of a

amino acid in the active site

– This binding may block the active site binding groups

so that the enzyme-substrate complex cannot form

– Vmax is decreased and no effect on Km

 • Divided into three categories:
1. Group specific:- reacts with specific R-
groups (side chain) of amino acid residues in
active site of enzyme.

•Di-isopropylphosphofluoride DIPF
Inhibit trypsin, chymotrypsin,
elastase and phosphoglucomutase

•Iodoacetate and heavy metals

like Pb, Ag, Hg etc react with –SH
group of cysteine residue present
at active site of enzyme
2. Substrate analogue:-
• Substrate analogue covalently react with amino acid of
active site of enzyme & permanently block the active
site.
• eg 3-bromoacetol phosphate (BAP) is analogue of
DHAP for enzyme phosphotriose isomerase

3. Suicide inhibitor:-
• The inhibitor binds to the active site where it is
modified by the enzyme to produce a reactive group
that reacts irreversibly to form a stable inhibitor-
enzyme complex.
• Some clinical examples of suicide inhibitors include:
• Aspirin, which inhibits cycloxygenase 1 and 2 enzymes.
• Penicillin, which inhibits DD-transpeptidase from building bacterial
cell walls.
• Allopurinol, which inhibits uric acid production by xanthine oxidase
in the treatment of gout.
• Azidothymidine and other chain-terminating nucleoside analogues
used to inhibit HIV-1 reverse transcriptase in the treatment of
HIV/AIDS.
• Sarin is a suicide inhibitor of acetylcholinesterase.
• 5-fluorouracil acts as a suicide inhibitor of thymidylate synthase
during the synthesis of thymine from uridine. This is often used in
combination with Methotrexate, a potent inhibitor of dihydrofolate
reductase enzyme.
• Exemestane, a drug used in the treatment of breast cancer, is an
inhibitor of the aromatase enzyme.
 Regulation of Enzyme Activity
One of the major ways that enzymes differ from
non-biological catalysts is in the regulation of
biological catalysts by cells
Enzyme quantity – regulation of gene expression
(Response time = minutes to hours)
a) Transcription(RNA made)
b) Translation (protein made)
c) Enzyme turnover
Enzyme activity (rapid response time = fraction of
seconds)
d) Allosteric regulation
e) Covalent modification
f) Proteolytic cleavage of proenzyme
 Allosteric Regulation
• Allosteric enzymes are regulated by molecules called
effectors (modifiers or modulators).
• It bind non-covalently at a second site (site other than
the active site).
• All allosteric effector can alter the affinity of the enzyme
for its substrate.
• Inhibit enzyme activity -----negative effectors.
• Increase enzyme activity-----positive effectors.
– Some effectors speed up enzyme action (positive allosterism)
– Some effectors slow enzyme action (negative allosterism)
 Feedback Inhibition
• In some pathway , end product inhibits the regulatory
enzyme of the pathway to regulate its own synthesis,
called feedback inhibition.

• In this example, product F serves to inhibit the activity

of enzyme E1
 Carbamoyl
phosphate Acetyl CoA
+
Aspartate
Aspartate
transcarbamoylase HMG CoA

Carbamoyl HMG CoA Reductase

Aspartate Mevalonate
+
Pi

Cytidine
triphosphate Cholesterol
(CTP)
Biosynthesis of pyrimidine
Biosynthesis of cholesterol
nucleotides
 Regulation of Enzyme Activity by
Covalent Modifications
• Usually regulatory enzymes are covalent modified
by their phosphorylation & dephosphorylation.
• Its catalyzed cyclic AMP dependent protein kinase
and phospho-protein phosphatase.
• A phosphate is transferred from an activated donor
(usually ATP) to an amino acid on the regulatory
enzyme
 CHAPTER 7

Regulatory enzymes (rate- limiting enzymes)

controlled by covalent modulation

Rafi M D: Textbook of Biochemistry (4 th

Edition) Universities
 Signaling Regulation of Glycogen
Synthase and Phosphorylase

A-forms, most active B-forms, less active

Induction & repression of Enzyme synthesis
•Cells can also regulate the amount of enzyme
present, usually by altering the rate of enzyme
synthesis.
•The increased (induction) or decreased
(repression) synthesis of the protein leads to an
alteration in the total population of active sites.
•For example
↑ levels of insulin due to high blood glucose
levels cause ↑ synthesis of key enzymes involved
in glucose metabolism.
Heme synthesis is regulated by repression of
transcription of structural gene of δ-ALA
 Proenzymes or Zymogen
• Some enzymes are synthesized and secreted in
inactive form which called proenzyme or
zymogen.
• It is converted to its active form before they act
on the substrate.
– By proteolysis (hydrolysis of the enzyme)
– Eg:
• Pepsinogen is synthesized and transported to the
stomach where it is converted to pepsin
Proenzymes of the Digestive Tract
 ISOENZYMES-ISOZYMES
 Isoenzymes are multiple forms of an
enzyme.
 Catalyze the same biochemical reaction.
 Tissue specific
 Isoenzyme patterns can give information
about organ-specific disease.
 Differ in chemical & physical properties
 AA sequence
 AA composition
 Structure
 Electrophoretic & immunological properties
 Kinetic properties (Vmax, Km)
 Degree of denaturation
 For example
• Lactate dehydrogenase (LDH)
• Creatine kinase (CK) or Creatine
phosphokinase (CPK)
• Alkaline phosphatase (ALP)
 LACTATE DEHYDROGENASE (LDH)

 It is a tetrameric protein and

made of
two types of subunits namely H (Heart),
M (skeletal muscle)
 Each encoded by a different gene.
 It exists as 5 different isoenzymes with
various combinations of H and M
subunits
 LDH is elevated in myocardial infarction.
Isoenzyme Composition Present in Elevated in
name

LDH1 ( H 4) Myocardium, myocardial

RBC infarction

LDH2 (H3M1) Myocardium,

RBC

LDH3 (H2M2) Kidney, Skeletal Malignancy and

muscle leukaemia

LDH4 (H1M3) Kidney, Skeletal

muscle

LDH5 (M4) Skeletal muscle, Muscular

Liver dystrophy and liver
diseases
LACTATE DEHYDROGENASE IN MI
 CREATINE KINASE (CK)
 Creatine kinase is a dimer made of 2 monomers
occurs in the tissues

 Skeletal muscle contains M subunit, Brain contains

B subunits

 Three different iso-enzymes are formed

Isoenzyme
Composition Present in Elevated in
name

CK-1 BB Brain CNS diseases

Myocardium/ Acute myocardial

CK-2 MB
Heart infarction

Skeletal
CK-3 MM muscle, myopathies
Myocardium
Alkaline phosphatase (ALP):-
Six isoenzymes

• The normal serum

levels of ALP range
between 40 and
125 U/ L.

• Increased ALP
levels are most
commonly
associated with
bone disease and
hepatobiliar
y disease.
Alcohal dehydrogenase (ADH):-
Two hetero dimer
American & Europeans αβ1
Japanese & Chinese αβ2
αβ2
Alcohal acetaldehyde (accumulation) tachycardia

Japanese & Chinese have increased sensitivity to alcohol.

 Diagnostic Enzymes
The levels of diagnostic enzymes in the
blood can be used to determine the
amount of damage in specific tissues
 Important enzymes in the investigation of
heart disease are CK, LDH and AST.
 Important enzymes in the investigation of
liver disease are AST, ALT.
 Alkalinephosphatase can be used in the
investigation of liver and bone disease.
 Increased levels of acid phosphatase are
found in prostate cancer.
 ALANINE TRANSAMINASE (ALT) AND
ASPARTATE TRANSAMINASE( AST)

- Oxoglutarate + L-aspartate - Oxoglutarate + L-alanine

Aspartate Alanine
aminotransferase (AST) aminotransferase
(ALT)
L- glutamate + oxaloacetate L - glutamate + pyruvate
 Alanine transaminase (ALT) and Aspartate transaminase (AST)
enzymes are the most abundantly present in the liver and is
elevated in blood as a result of leakage from damaged cells

 Alanine transaminase (ALT) increase is specific for liver

damage involving hepatocellular damage

 Aspartate transaminase (AST) is moderately increased in

Muscular dystrophy and acute myocardial infarction
LEVELS OF ENZYMES IN DISEASES
INVOLVING LIVER DAMAGE

In viral hepatitis
Rapid rise in
transaminases
(AST & ALT) in
serum occurs even
before bilirubin
rise is seen
LEVELS OF ENZYMES IN MYOCARDIAL
INFARCTION
NAME OF THE ENZYME Normal values PRESENT IN
Aspartate Amino transferase 8-20U/L Myocardial infarction
(AST) or (SGOT)

Alanine Amino transferase (ALT) 13-40U/L-Male Viral hepatitis

Or (SGPT) 10-28U/L- Female

Alkaline Phosphatase (ALP) 40-125U/L Various bone disorders,

obstructive liver diseases

Acid Phosphatase (ACP) 2.5-12U/L Metastatic carcinoma of the

prostate
 glutamyl Transferase ( GT) 0-30IU/L Various liver diseases

Creatine kinase (CK) 15-100U/L Muscle disorders and

myocardial infarction

Lactate Dehydrogenase (LDH) 90-200IU/L Myocardial infarction

 Amylase 50-120IU/L Acute pancreatitis

Use of Enzymes as Therapeutic Agents

Enzyme Use
Asparaginase Leukaemia
Collagenase Skin ulcers
Glutaminase Leukaemia
Hyaluronidase Heart attack
Lysozyme Antibiotic
Ribonuclease Antiviral
Dissolve blood clots &
Streptokinase
Myocardial Infraction
Trypsin Inflammation
Uricase Gout
Blood clots & pulmonary
Urokinase
embolism
 Enz ymes used in Laboratory
Measurements and Gene Transfer
 MCQs
1-The following is a group-specific enzyme:
A. Pepsin
B. Aminopeptidase
C. Phospholipase D
D. All of the above

2. The following is a substrate-specific enzyme:

A. Hexokinase
B. Thiokinase
C. Lactase
D. Aminopeptidase
3. The following is not a substrate-specific enzyme:
A. Glucokinase
B. Fructokinase
C. Hexokinase
D. Phospofructokinase

4. Chymotrypsin hydrolyses peptide bonds in which

carboxyl group is contributed by:
A. Phenylalanine
B. Tyrosine
C. Tryptophan
D. Any of the above
5. Coenzymes combine with:
A. Proenzymes
B. Apoenzymes
C. Holoenzymes
D. Antienzymes

6. Coenzymes are required in the following reactions:

A. Oxidation-reduction
B. Transamination
C. Phosphorylation
D. All of the above
7. The following coenzyme takes part in hydrogen
transfer reactions:
A. Tetrahydrofolate
B. Coenzyme A
C. Coenzyme Q
D. Biotin

8. The following coenzyme does not take part in

hydrogen transfer reactions:
A. FAD
B. NAD
C. NADP
D. Cobamides
9. The following coenzyme takes part in oxidation
reduction reactions:
A. Pyridoxal phospate
B. Lipoic acid
C. Thiamin diphospate
D. None of the above

10. In conversion of glucose to glucose-6-phospate, the

coenqyme is:
A. Mg++
B. ATP
C. Both of the above
D. Neither of the above
11. Enzymes having slightly different molecular structures
but performing identical activity are
a) holoenzymes
b)apoenzymes
c) isoenzymes
d) coenzymes

12. Which factor is responsible for inhibition enzymatic

process during feed back?
a) Enzymes
b) End product
c) Temperature
d) Substrate
14 The initial rate of an enzyme catalysed reaction depends on:
(a) the concentration of the enzyme
(b) the concentration of the substrate
(c) the affinity of the enzyme for its substrate
(d) all of the above
(e) none of the above
15 Kinase reactions:
(a) inhibit ATP breakdown
(b) involve the addition or removal of a phosphate group
(c) involve the addition or removal of a ketone group
(d) involve the addition or removal of an amino acid to a
polypeptide chain
(e) involve the transfer of hydrogen atoms
16 The energy for all forms of muscle contraction is provided by:
(a) ATP
(b) ADP
(c) phosphocreatine
(d) oxidative phosphorylation
(e) generated in the mitochondria of the cell
17Which of the following factors can affect enzyme activity?
(a) temperature
(b) pH
(c) the presence of certain metal ions
(d) the addition or removal of phosphate
(e) all of the above
18 Prosthetic groups are:
(a) required by all enzymes in the cell
(b) loosely bound to enzymes via hydrogen bonds
(c) sites on the enzyme molecule that permit allosteric modification of
enzyme activity
(d) linked to phosphate groups
(e) tightly bound to enzymes and are required for their activity
Proteolytic cleavage of
proenzyme(zymogen)
Proinsulin to Insulin
 Blood Clotting
•Clotting involves series of
X
zymogen activations
X
•Seven clotting factors are serine
X X
proteases involved in clotting
cascade rxns
X