Enzymes

Reaction
Dr. Yogi P. R.
Biochemistry Department
Medical Faculty
Swadaya Gunung Jati University
Cirebon 2009

Tujuan pembelajaran

Untuk memahami dan mampu menjelaskan

1.Pengertian, karakteristik, dan klasifikasi enzim
2.Fungsi Enzim dalam regulasi reaksi kimia
tubuh
3.Faktor-faktor yang mempengaruhi kecepatan
reaksi enzim dan regulasi aktifitas enzim
4.Fungsi intraseluler melalui regulasi enzim

DEFINITION & CHARACTERISTIC

Enzymes are :
- Proteins
- Metabolic catalysts
- The largest and most highly specialized catalysts
in the body for the reactions involved in metabolism
which increase the rate of chemical reactions by
lowering the activation energy of that reactions
- Unchanged number of enzyme before and after
reaction

E : Enzymes

ES : Enzymes+Substrates

S : Substrates

ES low stability

: Product

Enzyme Function

LOWERING ACTIVATION ENERGY

THE FUNCTION OF CATALYST

ENZYME IS A BIOCATALYST

Site of activity
A. Endoenzyme
Intracellular enzyme : ATP synthesis
B. Eksoenzyme
Extracellular enzyme

Catalysts effort

Occurred process
A. Constitutive enzyme
The number of enzyme always constant, not influence
by substrate concentration
B. Adaptive enzyme
The occurred process is stimulated by substrate

STRUCTURE OF ENZYMES

Cofactor :
Prostetic group
Coenzyme
Activator

COFACTOR COENZYMES
Thiamine pyrophosphate, from Vit. B1, Decarboxylase
Flavin mono/adenine di nuceotide, Vit. B2,
Dehydrogenase
Nicatinamide Adenine Dinucleotide/ Phosphate,
Nicotinic acid, Dehydrogenase
Coenzyme A, Panthotenic acid, Dehydrogenase
Pyridoxal phosphate, Vit. B6, Transferase
Tetrahydrofolic, Folic acid, Transferase
Deoxyadenosylcobalamine, Vit. B12, Isomerase

COFACTORS ACTIVATOR

Fe2+ or Fe3+ in Cytochrome oxidase,

Catalase and Peroxidase
Co in Dinitrogenase
K+ in Pyruvate kinase
Mg+ in Glucose 6-phosphatase

Interaction Enzyme-Substrates Model

Lock and key (1890 Emil Fischer)

Stereospecificity catalysts
The shape, or configuration, of the active site is
especially designed for the specific substrate involved
The configuration is determined by the amino acid
sequence of the enzyme, the native configuration of the
entire enzyme molecule must be intact for the active site
to have the correct configuration
The substrate then fits into the active site of the enzyme
in much the same way as a key fits into a lock

Induced fit (Daniel Koshland)

The binding of a substrate (S) by an enzyme is an interactive
process
The shape of the enzyme's active site is actually modified
upon binding S, in a process of dynamic recognition between
enzyme and substrate called induced fit
In essence, substrate binding alters the conformation of the
protein, so that the protein and the substrate "fit" each other
more precisely

Specificity Level of Enzymes

1. Bond specificity (Low specificity)

peptidase, phosphatase, esterase
2. Group specificity (Middle specificity)
hexokinase
3. Absolute specificity (High specificity)
urease

Velocity, Enzymes, Substrates

Acceleration of product is determined by enzyme concentration and

substrates concentration
V = Velocity
[E] = Enzyme concentration
[S] = Substrates concentration
A.If the S is CONSTANT The increase of V is equal with the increase
of E
B.If the E is CONSTANT and S increase V will increase proportionally
with the increase of S, but in higher concentration of S, the increasing
of V will decrease slowly until V was almost not suspended from S

A.

B.

Michaelis Menten Model

Leonor Michaelis & Maud Menten -1913

Konstanta Michaelis-Menten Km = k2+k3/k1

Km = The substrate at wich the velocity of the reaction is half the maximum
velocity
Km -- enzyme substrate complex high affinity
Km -- enzyme substrate complex low affinity

Factors that influence enzymatic reaction

1. Substrates
the substrates concentration will enzymatic reaction
until maximum condition
2. pH
optimum pH will enzymatic reaction
example : Amilase -- optimum pH 5,0
Arginase -- optimum pH 10

Substrate Concentration

Rate of Reaction

Active sites full- maximum turnover

Substrate Concentration

pH
Rate of Reaction

Narrow pH optima

Disrupt Ionic bonds - Structure

Effect charged residues at active
site

3. Temperature
optimum temp will enzymatic reaction
higher than optimum temp will damage enzyme
( 50C)
If you heat the protein above its optimal
temperature bonds break meaning the protein loses it
secondary and tertiary structure

Effect of heat on enzyme activty

Denaturing the protein

ACTIVE SITE CHANGES SHAPE
SO SUBSTRATE NO LONGER FITS
Even if temperature lowered enzyme

cant regain its correct shape

4.

Inhibitor

Competitive inhibitor
Another substance (analog substrates) has similar structure to
substrate
Succinate

Fumarate
Succinate
Dehydrogenase

Malonate

These compete with the substrate molecules for the active site
Always reversible
Increasing substrate concentration to against competitor

 Non-Competitive inhibitor
These are not influenced by the concentration of the substrate

It inhibits by binding irreversibly to the enzyme but not at the active site

Examples

Cyanide combines with the Iron in the enzymes cytochrome oxidase

Heavy metals, Ag or Hg, combine with SH groups.

 Feed-back inhibitor

The first step (controlled by eA) is often controlled by the

end product (F)
Therefore negative feedback is possible
A

eAC

eD
B

eEC

FeD

eF

Inhibition

The end products are controlling their own rate of

production

2008 Paul Billiet ODWS

 Alosteric inhibitor

These enzymes have

two receptor sites
One site fits the
substrate like other
enzymes
The other site fits an
inhibitor molecule

Substrate
cannot fit
into the
active site

Inhibitor
molecule

Inhibitor fits
into allosteric
site

Five Main Ways that Enzyme Activity is Controlled

in The Cell
1. Enzyme production (transcription and translation of enzyme
genes) enhanced or diminished by a cell in response to
changes in the cell's environment
This form of gene regulation is called enzyme induction and inhibition . For
example, bacteria may become resistant to antibiotics such as penicillin
because enzymes called beta-lactamases are induced that hydrolyse the
crucial beta-lactam ring within the penicillin molecule

2. Enzymes can be compartmentalized, with different metabolic

pathways occurring in different cellular compartments
For example, fatty acids are synthesized by one set of enzymes in the
cytosol, endoplasmic reticulum and the Golgi apparatus and used by a
different set of enzymes as a source of energy in the mitochondrion,
through -oxidation

3. Enzymes can be regulated by inhibitors and activators

This helps allocate materials and energy economically, and prevents
the manufacture of excess end products. The control of enzymatic
action helps to maintain a stable internal environment in living
organisms

4. Some enzymes may become activated when localized to

a different environment (eg. from a reducing (cytoplasm)
to an oxidising (periplasm) environment, high pH to low pH
etc).
For example, hemagglutinin in the influenza virus is activated by a
conformational change caused by the acidic conditions, these occur
when it is taken up inside its host cell and enters the lysosome

5. Enzymes can be regulated through post-translational

modification.
For example, in the response to insulin, the phosphorylation of multiple
enzymes, including glycogen synthase, helps control the synthesis or
degradation of glycogen and allows the cell to respond to changes in
blood sugar
Another example of post-translational modification is Chymotrypsin, a
digestive protease, is produced in inactive form as chymotrypsinogen in
the pancreas and transported in this form to the stomach where it is
activated. This stops the enzyme from digesting the pancreas or other
tissues before it enters the gut. This type of inactive precursor to an
enzyme is known as a zymogen.

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