ENYME

KINETICS
 ENZYME
INHIBITION
INHIBITORS
- chemicals that reduce the rate of enzymatic reactions
- block the enzyme but they do not usually destroy it
- usually specific and work at low concentrations

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 Two Classes of
Inhibitors in the Extent
of Interaction
1 . Irreversible Inhibitors
-combine with the functional groups of the amino acids in the active site,
irreversibly.

Examples: nerve gases, pesticides

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 Two Classes of
Inhibitors in the Extent
of Interaction
2. Reversible Inhibitors
- The site of attack is an amino acid group that participates in the normal enzymatic
actions in the extent of interaction. Forms covalent or very strong non covalent
bonds.

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TYPES OF REVERSIBLE INHIBITORS

Non- Reversible
co Inhi
mp bitor
etiti s
ve
inhi
bito
r Competitive
inhi
bito
r

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Competitive inhibitor
These compete
with the substrate
molecules for the
active site.

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Non- competitive inhibitor
Not influenced by
the concentration
of the substrate.
Inhibits by binding
irreversibly to the
enzyme but not at
the active site

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 COMPETITIVE AND NON-
COMPETITVE

Non- competitive
Competitive • Inhibitor does
• Inhibitor not bind at the
competes with active site
the substrate at
the active site • Active site may
• Active site stays change shape
the same (e.g. allosteric
inhibition)

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 Models for more
Complex Enzyme
Kinetics
Allosteric
enzymes
kinetics
the binding of one substrate to the enzyme
facilitates binding of other substrate molecules.

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Allosteric
enz
ym
es
kin
etic
s
Allosteric
enzymes
– a class of enzymes that bind
small, physiologically important
molecules and modulate activity
in ways

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 Kinds of Allosteric
Enzymes

1. Positive
–activates enzymes
2. Negative
– deactivates enzymes

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Enzyme
Activators


compounds that increase Example:

enzymatic activity. fructose 2,6-


are usually involved in biphosphate

allosteric enzymes for
metabolism regulation

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Factors affecting
Enzyme Activity
Temperature
As the temperature increases, the rate of reaction of enzyme also increases. But
very high temperatures denature enzymes.

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pH Different enzymes work best at different pH values. The optimum pH for an
enzyme depends on where it normally works.

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Other factors:
•

Metal/ Salt Concentration – each enzyme has an

optimal salt concentration.
•

Concentration of the Substrate – As the

concentration increases, the enzyme reaction rate
increases.
•

Concentration of Enzyme – increasing enzyme

concentration will increase the enzyme reaction rate.
•

Steric Hindrance – because of the “separation” or

spacing, the substrate is very difficult to bond with the
enzyme in the active site.
 Enzyme
Immobilization
Enzyme
Immobilization
-restricts the mobility of an enzyme or protein and fixes the
enzyme into a state without disturbing its functional ability

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 Methods of
Immobilization
 1.
Adsorption
-Enzyme is
adsorbed on the physical
outer surface of the
support. It can affect the
functional ability of
enzyme by blocking its
active site.
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Carriers used in adsorption can be

Mineral-
Carriers
bas Organic
ed Bi
sup m
por ol
t ec
ul
ar Modified
ba io
se n
d ex
su ch
pp an
or ge
t re
sin 24
Absorptive immobilization of enzymes

Static Method
- Enzyme is
immobilized by Dynamic Method
allowing it to be - This process typically
in contact with involves the
the carrier admixing of
without enzyme with
agitation. the carrier
under constant
agitation using
mechanical
shaker.

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Absorptive immobilization of enzymes

Reactor loading Method

- The carrier is placed into
the reactor and
Electro-deposition
enzyme solution
Method
is transferred to
- carrier is placed in the
the reactor with
vicinity of an
agitation of the
electrode and
whole content in
the enzymes
the reactor.
migrate to
carrier in
presence of
electric current.

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2.
Covalent
Bonding
-The method utilizes
chemical groups present
on both enzyme and
carrier for immobilization.

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Carriers used in covalent bonding can
be

Carriers

Biomolecules

Protein
ca
rri
er Inorganic
Synthetic s m
m ol
ol ec
ec ul
ul es
es
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Covalent bonding can be done by

Peptide Bond
Diazoation

Polyfunctional
agent

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3.
Entrapment
- Entrapment is carried
out by mixing the biocatalyst
into a monomer solution,
followed by polymerization
initiated by a chemical
reaction.

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 Methods of
entrapment
1. Inclusion in the Gel – enzyme is trapped inside
the gel, which is formed by the polymer.
2. Inclusions in fibers – enzymes are supported
on the fibers of the supporting material forming
the matrix.
3. Microcapsules – enzymes are trapped in the
microcapsules. Most common microcapsules are
polyamines and sodium alginate

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 4.
Cross-linking
process /
Copolymerization
-This method is based on
the formation of covalent
bonds between the enzyme
molecules, by means of
multifunctional reagents,
leading to three
dimensional cross linked
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aggregates.
 5.
Encapsulation

- An enzyme is
encapsulated within a
capsule made up of semi-
permeable membrane like
nitrocellulose, nylon and
hemi-cellulosic structures.
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Effect of Mass
Transfer
Resistance
•

Immobilization of an enzyme transforms a

homogeneous (soluble) catalyst into a heterogeneous
(insoluble) system.
•

Carrier binding techniques introduce external mass

transfer effects between the liquid phase and the solid
surface.

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•

Immobilization of an enzyme transforms a

homogeneous (soluble) catalyst into a heterogeneous
(insoluble) system.
•

Carrier binding techniques introduce external mass

transfer effects between the liquid phase and the solid
surface.

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 An enzyme immobilized through binding to a carrier bead and placed in a simple flow
may be represented by the following illustration.

The change in concentration of a reagent A from

[A]bulk to [A]surface takes place in a narrow fluid
layer next to the surface of the sphere.
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 N A  kc Ap ([ A]s  [ A])

NA = transfer rate: mole/s

•

kc = convective mass transfer coefficient: m/s

•

AP = surface area of the particle: m2

•

[A] = concentration of solute at the surface and in the

bulk,respectively: mole/m3

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Electrostatic
and Steric
Effects
Shift in the pH-activity profile is given by

pHi= Internal pH value

pHe= External pH value
Z= charge (valence) on the substrate
Nf= 96 500 coulumb/ eq.g (Faraday Constant )
Ψ = Electrostatic Potential
R= gas constant

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 Industrial
Application of
Enzymes
1.
Industrial
Production
- commercial production
of antibiotics, beverages,
amino acids and secondary
metabolites of industrial grade

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2.
Biomedical
Applications
– commonly used in the fast
diagnostic kits like ELISA and
treatment of many pathogenic
diseases

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3.
Food Industry

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Food Industry
• Pectinases and • Amylases from fungi
Cellulases and plants - production
– used in the production of of sugars from starch.
jams, jellies and fruit Such as in making high-
and vegetable fructose corn syrup.
syrups • Lipases
- is implemented during the
• Lactase production of Roquefort
immobilized with cheese to enhance the
cellulose fibers ripening of the blue-
– produces lactose-free milk mold cheese.
• Papain
-to soften meat for cooking

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4.
Biodiesel
Production

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 5.
Waste water
Management
– treating sewage and industrial effluents
using packed bed reactors

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6.
Starch Industry
Amylases,
amyloglucosideases and
glucoamylases - converts starch
into glucose and various syrups.

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 7.
Brewing
Industry
– Enzymes frm barley are released during the
mashing stage of beer production. They
degrade starch and proteins to produce
simple sugar, amino acids and pepticides
that are used by yeast for fermetatation.
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8.
Paper Industry
•

Amylases, Xylanases, cellulases

and ligninases - degrade starch to lower
viscosity, aiding, sizing and coating paper
•

Xylanases reduce bleach required for

decolorizing;
•

cellulases smooth fibers, enhance

water drainage and promote ink removal,
•

lipases reduce pitch and lignin-

degrading enzymes remove lignin to
soften paper. 50
 9.
Biofuel Industry
•

Cellulases - used to breakdown

cellulose into sugars that can be
fermented

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 10.
Biological
Detergent
•

Primarily proteases - produced in an

extracellular from bacteria and used for
presoak conditions and direct liquid
applications helping with removal of
protein stains from clothes.

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 11.
Rubber Industry
Catalase - to generate oxygen from
peroxide to convert latex into foam rubber

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 12.
Photographic
Industry
Dissolve gelatin oil scrap film, allowing
recovery of its silver content.

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 Activity
•

Unwind the words!

ut your brain to the test by unscrambling words
and then using the meaning to solve the riddle.
hose who will participate and
uess the correct answer
ill receive a plus point in quiz.
srotbihiin

- chemicals that reduce the rate of enzymatic

•

reactions and block the enzyme

•

but they do not usually destroy it

Reapeutrmet

- one of the factors affecting enzyme activity-

as it increases, the rate or reaction of enzym
e also increases
Ozantioaid

- the reaction occurs between amino group of

the carrier and Tyrosil and Histidyl group of th
e enzyme
Lypoiremazniotco

- this method is based on the formation of co

valent bonds between the enzyme molecule,
by means of multifunctional reagents, leading
to three dimensional crossed linked aggregat
es.
Uspacalneniot

- the effectiveness depends on the stabilityof

the enzyme inside the capsule
Ainpap

- to soften meat for cooking

Selasulelc

- used to break down cellulose into sugars th

at can be fermented
Mezeny iotnvaitca

- compounds that increase enzymatic activity

 GROUP 7
ChE 4101

DE CASTRO, NICA MARIEL

LOPEZ, AUBRENICA ROSE PAULINE
UMALI, JONALYN
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