Enzyme

Prof Bagiada
CBM team

Gene structure and its

biological activity
Gene encode the information that
determine the structure of
protein.
In turn the protein structure
determine its biological activity.
Through the biological activity of
protein the cells physiological role
is determined.

Functional group of protein

Protein Is the most abundant organic mol
of the cells
Why is so abundant and vital for the cells:
(great variety and function of protein in
the physiological activity of the cells)
Protein are capable assuming so many
roles because their structure can vary
enormously, depend on the variation of aa
sequence of the molecule
Protein can act as : Enzyme, Receptor,
Transport, Immunologic, etc

Definition
Enzymes : Specific protein catalyzed
for chemical reactions
Why it is specific ?
Catalyze: protein catalyze and non
protein catalyze.
Non protein catalyze: will catalyzed
many kind of reaction (H+, OH-, Cu+
+, etc)

Nomenclature and
classification
Enzyme : En=In, Zyme =yeast
Firstly : depend on the founder (ptyalin)
Then : ase, after substrate
(amylase, lipase, protease)
IUBS :
1.Enzyme classified into 6 mayor class.
(1. oxido reductase, 2 transferase, 3
Hydrolase, 4. lyase, 5. Isomerase
6.ligase)

2. The enzymes name consist 2 part : the

first is the substrate, second part ending
by ase which confirm type of reaction
catalyzed (glutamat dehydrogenase)
3. Additional information is needed to
clarify the nature of the reaction, may
follow in parenthesis (There trivial
name.)
4. Systematic code number= Enzyme Code
(E.C) consist of 4 digit (EC 2.7.1.1)

Enzyme constituent
Enzyme : apo-enzyme and co-enzyme
holo enzyme
Coenzyme : non protein part of the
enzyme, which is partly resistance to
heating, can be dialyzed, frequently
contain of B vitamin as part of their
structure.
Classification of coenzyme:
1. CoE that transfer H+
2. CoE that transfer non H

 CoE for transfer of H:1. NAD+,

NADP+,2. FMN, FAD 3.Lipoic acid, 4.
Co.Q
CoE for group transfer other than H :
1. ATP and its relative, 2.Sugar
phosphate, 3.CoA,
4.Thiaminpyroposphate, 5.B6phosphate, 6. Folate coenzyme, 7.
Biotin 8.Cobamid CoE, 9 Lipoic acid

Factors influencing enzyme

reaction
General reaction of enzyme.
E + S ES E + P
1. Substrate concentration
2. Enzyme concentration
3. Temperature
4. Acidity (pH)
5. Inhibition
6. Oxidation

Substarte concentration
V

vmax----------------------- V max----------------is very much

than Km

K
m

The Michaelis Menten expression

[S]/Km + [S]

Km = Constanta Michaelis

When concentration S
less than Km.
When S is very much greater
When concentration S = Km
v=V

Temperature
Temp. optimum
V

Temperature
(0 C)

37

70

 pH values between 5.0 and 9.0

A few enzyme such as pepsine
optimal pH <2
The shape of pH activity curve is
determined by : 1. Enzyme at
extremely high or low pH
2. Effect on the charged state of
the S.

Inhibition of an Enzyme
Two broad classes of inhibition:
1. Competitive inhibition
2. Non competitive inhibition
Competitive inhibition=substrate analog
inhib.
E + S ES E + P

Enz

EnzI

EnzS

Enz + P
Enz + P

 Competitive inhibitors that block

enzyme reactions in a parasite are
potent chemotherapeutic agents:
Sulfanilamide, structural analog of
PABA will block folic acid synthesis.
Folic acid is important for bacteria to
divided
Pteridine + PABA + Glutamic acid
FA

 Folic acid analog used as

chemotherapeutic agent again tumors :
aminopterin (4-amino folic acid) C-I of
dihidrofolate.
amethopterin(MTX) is C-I of dihidrofolate
in dihidrofolat reductase reaction.
Antagonist to B vitamin : pyrithiamin and
oxythiamin antagonist to B1.
Sulfonamide derivatives acetazolamid
(Diamox) is C-I of Carbonic anhydrase

Non Competitive InhibitionNCI

NCI : reversible and irreversible NCI
Reversible NC:
ENz +/- I EnzI + S ENzIS E
+ P
Enz +/- S EnzS I EnzSI E
+ P

 Irreersible NCI
A wide of enzyme poison such as
Iodoacetamide, heavy metal (Hg, Ag)
Oxidizing agent reduce enzyme
activity

ISOZYMES
Isozyme = iso enzyme
Are distinct form of the same catalytic
activity.
Medical interest in isozymes start in
1957: where human sera contained
several lactate de-hydrogenase
isozymes.
LDH isozymes differ from one another at
the level of quarternary structure

 The active of lactate dehydrogenase

mol consist of 4 subunits of 2 types H
and M
Only the tetrameric mol possesses
catalytic activity HHHH, HHHM,
HHMM, HMMM, MMMM. (I1, I2, I3, I4,
I5)

CK/CPK
CREATINE KINASE
3 ISOZYM: CK-MM, CK-MB, CK-BB
CK-MB MOST PROMINENT IN AMI
M=Muscle B=Brain

Enzymes in clinical
diagnosis
A. Functional plasma enzyme
B. Nonfunctional plasma enzyme.
Ad A)
Are present all time in the circulation.
Level activity in plasma always high
Its substrate are in the blood.
Eg lipoprotein lipase, pseudocholin
esterase, enzyme for blood clot and blood
clot dissolution

 Ad B)
Non functional plasma enzyme.
No physiologic function in blood
Their substrate frequently absents from
plasma.
Its made in tissue/organ.
Its level activity in plasma much lower than in
the tissue.(10-6)
Elevated its activity in plasma, suggest as
increase tissue destruction

 For example enzymes in exocrene

secretions, and the true intracellular
enzyme.
Lipase :low in liver disease, vit A deff,
some malignancy and DM. Elevated in
acute pancreatitis, pancreatic Ca.
Amylase : Low in liver disease, and
increase in high intestinal obstructions,
parotitis, acute pancreatitis and DM

 Trypsine : elevated in acute disease of

pancreas. More sensitive dan lipase and
amylase antithrombotic
Cholinesterase: low level found in liver
disease, malnutrition and chronic debilitating,
and acute infectious disease and anemia.
High level occurs in the nephrotic syndrome.
Some insecticide (phosphate inorganic)
depress cholinesterase activity. Test of this
enzyme in blood used as indicator for over
exposure to insecticide

ALKALINE PHOSPHATASE
Alkaline phosphatase: capable of catalyzing the
hydrolysis of phosphate ester at alkaline pH.
Increase in rickets, hyperparathyroidsm, Pagets
disease, osteoblastic sarcoma, obsrtuctive
jaundice and metastatic Ca, conghestive heart
failur e (AS A RESULT TO THE LIVER)
Its isozyme are present in body fluids and
originate from bone, liver, placenta and
intestine
Have a great value to distinguish liver lesion
and bone lesion in case of metastatic Ca.

ACID PHOSPHATASE
CATALYSIS THE HYDROLISIS OF
PHOSPHATE ESTER IN ACID pH
Increase in metastatic prostatic Ca.

TRANSAMINASES
TWO IMPORTANT TRANSAMINASES
SGPT=ALT
SGOT=AST
SGOT: Capable to catalyze transfer of amino group of
aspartic acid to alpha keto glutaric acid forming
glutamic and oxaloacetic acid
SGPT: transfer the amino group of alanin to alpha
ketoglutaric acid forming glutaric acid and pyruvic
acid.
Heart muscle rich in transaminase enzyme AMI are
followed by rapid and stricking increases in serum
transaminase level decrease towards normal
within a few days(GOT/AST
Liver tissue both rich in Ast and ALT, but ALT is more
spesific indicator. Extensive skeletal muscle damage,
elevate transaminases

HOW NFPE INCREASE?

3 THEORIES
1. NECROTIC THEORY
2. CELL MEMBRANE LEAKAGE
3. IRRITATION AND PROLIFERATION
THEORY