BY

Enzymes
Tarachand nayak &
Umesh kr prajapat
CVAS,Bikaner Pepsi
n

30/04/10  
What are enzymes?
 Enzymes are proteins (globular).
 Act as organic enzymes.
 Enzymes speed up the rate of a chemical
reaction without themselves being used up
(consumed) during the reaction.
 Are essential to the functioning of all cells.
Without enzymes, metabolism would be too slow
& insufficient energy would be available to
maintain life.

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Enzymes are the proteins
that provide function to the
cell.

Enzymes are catalysts:

they speed up chemical
reactions.
Image

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 Name of Enzymes

 End in –ase
 Identifies a reacting substance
sucrase – reacts sucrose
lipase - reacts lipid
 Describes function of enzyme
oxidase – catalyzes oxidation
hydrolase – catalyzes hydrolysis
 Common names of digestion enzymes still
use –in
pepsin, trypsin

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 Classification of Enzymes

Class Reactions catalyzed

 Oxidoreductoases oxidation-reduction
 Transferases transfer group of atoms
 Hydrolases hydrolysis
 Lyases add/remove atoms
to/from a double bond
 Isomerases rearrange atoms
 Ligases combine molecules
using ATP

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 Examples of
Classification of Enzymes
 Oxidoreductoases
oxidases - oxidize ,reductases – reduce
 Transferases
transaminases – transfer amino groups
kinases – transfer phosphate groups
 Hydrolases
proteases - hydrolyze peptide bonds
lipases – hydrolyze lipid ester bonds
 Lyases
carboxylases – add CO2
hydrolases – add H2O

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 CHEMICAL NATURE OF
ENZYMES
 HOLOENZYME=APOENZYMES+COENZYMES
(active en.) (protein part) (non protein)

WORKING
OF ENZYMES

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SPECIFICTY OF ENZYMES

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Factors affecting enzyme
activity.
Enzymes are very sensitive to conditions in
which they work. They usually have a narrow
range of conditions under which they operate
properly.
 Factors affecting enzyme activity
Temperature.
pH.
Enzyme concentration.
Substrate concentration.
Enzyme inhibitors

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Factors affecting enzyme
activity.
Temperature
Enzymes usually work best in the temperature of
the environment in which they are found in.
e.g. most human enzymes have an optimum
temperature of ~37ºC (= body temperature).
At high temp enzymes are permanently
denatured.
At low temp enzyme activity slows down,
however no permanent damage is done to the
enzyme.

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Factors affecting enzyme
activity.
Temperature

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Factors affecting enzyme
activity
pH
Most human enzymes have an optimum pH
between 6 and 8. e.g. Trypsin has an optimum pH
of 8.0.
Enzymes that are secreted into the stomach have
a much lower optimum pH. e.g. Pepsin has an
optimum pH of 1.5.
If pH is too high or low, enzyme can become
denatured.

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Factors affecting enzyme
activity
pH

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Factors affecting enzyme
activity.
Enzyme concentration
 Rate of reaction
continues to increase
with an increase in
enzyme
concentration
(assuming non
limiting amount of
substrate)

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Factors affecting enzyme
activity.
Substrate concentration
 Rate of reaction
increases up to a
point. After this rate
levels off because all
enzyme molecules
are working at their
maximum capacity
(i.e. active sites are
saturated).

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Factors affecting enzyme
activity.
Enzyme inhibitors
 Two types
Competitive inhibitors – block the active site and
stop the substrate from getting in there.
e.g. poisons such as arsenic
Non-competitive inhibitors – bind to enzyme (but
not the active site) and cause it to change shape.

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 Factors affecting enzyme
action
Substrate concentration
Max. Rate

Rate of reaction

Substrate 
conc.
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MICHEALIS – MENTON EQUATION
Vi = V max [S]
Km + {S}
Vi = Measured initial velocity
V max = Maximum velocity
S = Substrate
Km = Michaelis constant
Variations
A. When (S) is much less than Km
Vi = V max [S] OR V max [S] K [S]
Km + {S} Km
So Vi depends upon substrate concentration

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 ENZYME INHIBITION
 Competitive inhibition
 Non competitive inhibition
 Irreversible inhibition
Competitive inhibition
 Inhibitors resemble substrate, Km is increased no
change in Vmax
 Succinate Enz Fumarate
 Malonate (structural analog of Succinate ) Enz –
inhibition
no product
 Drug Allopurinol, structural analog of Xanthene is used
for treatment of gout /hyperuricemia as it is a
competitive inhibitor of enzyme Xanthene oxidase which
normally converts Xanthene into Uric acid
 Addition of excess of normal [S] will reverse this
inhibition
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 ENZYME INHIBITION

NON COMPETITIVE INHIBITION

 Inhibitor binds on separate site on enzyme
therefore no competition with substrate. Vmax is
reduced and no change in Km
 Inhibitor can bind with either free enzyme or
enzyme – substrate complex and in both cases
render these inactive
 Lead poisoning is an example of this inhibition
and it inhibits enzyme Ferrochelatase which
adds iron molecule to the centre of porphyrin
ring in the synthesis of Hemoglobin
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 IRREVERSIBLE INHIBITION
Permanent covalent linkage with enzyme rendering it
irreversibly inhibited
 Diisopropyl phospho fluoride (DIPF)
 Iodoacetamide
 Heavy metal [Ag+ Hg+2], Silver, Mercury
 Oxidizing agents
 Covalent linkage with enzyme: inactivation of
enzyme
 Kinetics are same as of non competitive inhibition,
therefore difficult to distinguish between the two
 Examples are insecticides which act as enzyme
poisons for the insects & disinfectants used for
micro-organisms
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 CO-FACTORS OF ENZYMES
ENZYMES CO FACTORS
Catalase Iron
Peroxidase Fe2+ or Fe3+
Cytochrome oxidase

Cytochrome oxidase Copper : Cu+2

Carbonic anhydrase alcohol dehydrogenase Zinc : Zn2+
Hexokinase Magnesium
Glucose-6-phosphatase Mg2+
Pyruvate kinase
Arginase Manganese Mn2+
Pyruvate kinase Potassium K+
Urease Nickel N 2+
Glutathione
30/04/10 Peroxidase  Selenium : Se
 COENZYMES
 Heat stable, low mol wt organic compounds
non-covalently linked with enzymes can be
separated. APO + CO = Holoenzyme
 If covalently Linked to apoenzymes =
prosthetic group
 Act as intermediate or ultimate acceptor in
group transfer

D-G + Enzyme
A A-G + D
Co-
Enzyme

D A
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COENZYMES
CO ENZYMES COENZYMES FOR TRANSFER
FOR TRANSFER OF OTHER GROUPS
OF H+
NAD, NADP SUGAR PHOSPHATES
FMN, FAD THIAMINE PYROPHOSPHATE
TPP, PYRIDOXAL PHOSPHATE
LIPOIC ACID FOLATE AND COBAMIDE (VIT
B12), BIOTIN
COENZYME, Q LIPOIC ACID

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 CO-ENZYMES
REDUCTION OF NAD+ TO NADH.H+
Lactic acid + NAD LD Pyruvic acid + NADH-H+
H
Malic acid + NAD Oxalo
Malic dehydrogenase acetic acid +
NADH -H+
Glucose-6-phosphate + NADP 6-Phosphoglucon-
olactoneG-6-
+NADPH-H+
P.D
REDUCTION OF FAD OR FMN TO FADH2 OR FMNH2
FMN is co enzyme for Cytochrome C oxidase, L.Amino
acid dehydrogenase
FAD is co-enzyme for xanthene oxidase acyl-CoA
dehydrogenase

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 CO-ENZYMES
Thiamine pyrophosphate:
Co-enzyme for oxidative decarboxylation for ketoacids
CoANAD NADH-H+

Pyruvate Pyruvate Acetyl

dehydrogenase
CoA

Pyruvate decarboxylase
Pyruvate +TPP Acetalaldehyde -TPP
complex+Co2 -ketogluteratedehydrogenase
NAD NADH-
Alpha ketogluterate+6 CoA-SHH+ Succinyl
CoATransketolase
+ Co2
Ribose-5 Po4 + Xylulose-5-Po4 Sedoheptulose 7-Po4 +
3 30/04/10  
phosphoglyceraldehyde
CO-ENZYMES
Biotin
 Part of multiunit enzymes causing carboxylation reactions. Acts as
carrier of CO2
Acetylcarboxylase
Enz-Biotin-COO-  Enz-Biotin
Acetyl CoA+HCo3 + ATP Malonyl-CoA

Pyruvate carboxylase
.Biotin
Pyruvate+ HCo3 + ATP Oxaloacetate+
ADP+Pi Carbamoyl Po4.Synthetase -
Biotin

NH4 + HCo3 + 2ATP CarbamoylPO4 +

2 ADP+ 2 Pi

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Synthesis of Purines and Pyrimidines
 CO-ENZYMES
Ascorbic acid (Vitamin C)
 Strong reducing agent
 Required for hydroxylation of proline into hydroxyproline for
synthesis of collagen
Conversion of tyrosine into dopamine and into catecholamines
(adrenaline and noradrenalin)
Bile acid formation
Conversion of cholesterol into 7-hydroxylcholesterol
Maintain metallic co-factors like Cu+ in Monooxygenases and Fe in
dioxygenases in reduced form
Conversion of cholesterol into steroid hormone in adrenal cortex
Absorption of iron by reducing into reduced form which is can be
easily absorbed
Acts as antioxidant in GIT by preventing formation of nitrosamines
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during digestion  
CO-ENZYMES
 Folic acid
 Active form is tetrahydrofolate which acts as single
carbon carrier for synthesis of various compounds like
pyrimidines and purines e.g. conversion of dUMP
(deoxyuridylate) into dTMP (deoxythymidylate)

 Vitamin B12
Acts as co-enzyme in groups rearrangements in
isomerases e.g. conversion of methyl malonyl CoA into
succinyl-CoA by enzyme methylmalonyl-CoA mutase
Converts homocystein into methionine
Act
30/04/10 as maturation
  factor for RBCs
 Action of enzyme
(Anabolic reaction)
enzyme­substrate  enzyme­product 
complex complex

product
substrate

enzyme enzyme
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 Action of enzyme
(catabolic reaction)
enzyme­product  enzyme­substrate 
complex complex

products substrate

enzyme enzyme
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 Lock and key hypothesis

product

Substrate
product

Enzyme
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 Lock and key hypothesis

E S  
P
A T 
SH ’
O N
D C H
A T
M

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 Enzyme Action:
Induced Fit Model
 Enzyme structure flexible, not rigid
 Enzyme and active site adjust shape to
bind substrate
 Increases range of substrate specificity
 Shape changes also improve catalysis
during reaction

3838
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 Enzyme Action:
Induced Fit Model

P
S
S
P
E + S ES complex E + P

3939
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ENZYME SUBSTRATE BINDING AND
SPECIFICITY

 COMPLEMENTARITYO
F STRUCTURES

 STERIC, CHARGE
NEUTRALITY AND
HYDROGEN BOND
FACTORS

 ATP TO MYOSIN, Kd =
10-13 M

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REGULATION OF ENZYME
ACTIVITY
There are six different ways
1.Allosteric inhibition
2.Activation of latent enzyme(Proenzyme)
3.Compartmentation of metabolic pathway
4.Control of enzyme synthesis
5.Enzyme degradation
6.Isoenzyme

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ALLOSTERIC REGULATION

 Low molecular wt allosteric effectors structurally

not similar to substrate
E1 E2 E3
A B C D
Bind at sites other than active site leading to
feed back inhibition
Usually product or last small molecule before
macromolecules in biosynthesis

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PROENZYMES
 Inactive enzymes initially secreted as large molecules, active
site not exposed
Pepsinogen HCl Pepsin
Prochyomotrypsin Proteolysis Chymotrypsin

1 245 1 245
Trypsinogen
Trypsin
Enteropeptidase
1 15 16 245
π 7 245
Chymotrypsi Trypsin active form
n
1 16 146 149 Chymotrypsin active
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13 245
PROENZYMES
 Required for control of catalytic activity of
enzymes so that catalytic activities only occur
when required
 Pancreatic enzymes if all the time active  auto
digestion of pancreas
 Blood clot lysis enzymes only active when blood
clot is formed

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COMPARTMENTATION
 The synthetic(anabolic) & breakdown
(catabolic) pathways are operative in
different cellular organelles to achieve
maximum economy. For instance,
enzymes for fatty acid synthesis are
found in the cytosol where as enzymes
for fatty acid oxidation are present in the
mitochondria.

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CONTROL OF ENZYME
SYNTHESIS
 Synthesis of enzyme(protein) is
regulated by genes.

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ENZYME DEGRADATION
 There is a lot of variability in the half
lives of individual enzymes, enzyme with
long half life is usually sluggish in its
catalytic activity.
 In general, the key & regulatory enzyme
are most rapidly degraded.

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 CREATININE KINASE (CK): 3
ISOENZYMES
CK1 BB OCCURS IN 
BRAIN,SMOOTH 
MUSCLES of GIT AND 
URINARY TRACT

CK2 MB MYOCARDIUM (35 
%), SK MUSCLE (5%) 
  IN ACUTE MI
CK3 MM CK3 MM IN SK 
MUSCLES 
CREATININE KINASE
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LACTATE DEHYDROGENASE: 5
ISOENZYMES
LDH 1 HHHH Occurs in myocardium (aerobic
tissues)  in acute MI
LDH 2 HHHM  In acute leukemia

LDH 3 HHMM  In acute leukemia

LDH 4 HMMM Occurs in muscle and liver

(anaerobic tissues)
LDH 5 MMMM Occurs in muscle and liver
(anaerobic tissues)  in liver
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diseases
ENZYME PATTERN IN
DISEASES
 Enzymes in myocardial infarction
The enzymes namely creatine
phasphokinase(CPK),aspartate
transaminase(AST) & lactate
dehydrogenase(LDH) are important in the
diagnosis of MI.

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 DIAGNOSTICALLY IMPORTANT
ENZYMES
ENZYMES PRINCIPAL PRINCIPAL
SOURCE CLINICAL USE
ACID PROSTATE, CARCINOMA OF
PHOSPHATASE RBC PROSTATE
ALT (ALANINE LIVER, SK HEPATIC
TRANSAMINASE) MUSCLE, PARENCHYMAL
HEART DISEASE

AST (ASPARTATE HEART, LIVER, MYOCARDIAL

TRANSAMINASE) SK MUSCLE, INFARCTION,
KIDNEY, RBCs HEPATIC
DISEASE

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DIAGNOSTICALLY IMPORTANT
ENZYMES
ENZYMES PRINCIPAL PRINCIPAL
SOURCE CLINICAL USE
ALDOLASE SK MUSCLE, MUSCULAR
HEART DYSTROPHIES
ALKALINE LIVER, BONE, BONE,
PHOSPHATASE INTESTINE, HEPATOBILIARY
PLACENTA, DISEASE
KIDNEYS

AMYLASE SALIVARY PANCREATIC

GLANDS, DISEASE
PANCREAS,
OVARIES
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DIAGNOSTICALLY IMPORTANT
ENZYMES
ENZYMES PRINCIPAL SOURCE PRINCIPAL CLINICAL
USE
CREATINE KINASE SK MUSCLE, HEART, MYOCARDIAL
BRAIN, SM-MUSCLE INFARCTION MUSCLE
DISEASE

CHOLINE LIVER RGANOPHOSPHORUS

ESTRASE INSECTISIDE
POISONING, HEPATIC
DISEASE

GAMA-GLUTAMYL LIVER, KIDNEYS ALCOHALIC

TRANSFERASE HEPATOBILIARY
DISEASE

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DIAGNOSTICALLY IMPORTANT
ENZYMES
ENZYMES PRINCIPAL PRINCIPAL CLINICAL
SOURCE USE
LACTATE DEHY- HEART, LIVER, MYOCARDIAL
DROGENASE SK MUSCLE, INFARCTION,
RBCs, HEPATIC DISEASE
PLATELETS,
LYMPH NODES

GLUTAMATE DE- LIVER, HEPATIC

HYDROGENASE PARANCHYMAL
DISEASE

ISOCITIRIC DEH- LIVER --DO--

YDROGENASE
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DIAGNOSTICALLY IMPORTANT
ENZYMES
ENZYMES PRINCIPAL PRINCIPAL
SOURCE CLINICAL USE
5-NUCLEOTIDASE LIVER HEPATOBILIARY
DISEASE
GLUCOSE-6- LIVER HEMOLYTIC
PHOSPHATE DE- DISORDERS
HYDROGENASE

LIPASE PANCREAS ACUTE

PANCREATITIS
SORBITOL DEH- LIVER LIVER
YDROGENASE PARENCHYMAL
30/04/10   DISEASE
 Enzyme in muscle disease
increased level of CPK, aldolase AST.
 Enzyme in liver disease
1.Alanine transaminase
2.Aspartate transaminase
3.LDH

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 Enzymes in cancer
increase in the serum acid phosphatase is
specific for the detection of prostatic carcinoma.

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 thousands of
diseases

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Enzymes are required to
sustain
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health.  
Coolege of veterinary & animal sciences, Bikaner

Thankyou
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