Chapter 4

Reaction Mechanism
in Chain
Reaction
and

Biological Reactions
Part 2
Kinetics of Biological Reactions

By: Cik Siti Khatijah Jamaludin, FKK UiTM Shah Alam

 Subtopic covered in Chapter 3 (Part 2)
Enzymatic Reaction Fundamentals
(Chapter 7 Fogler, page 394-403)
Enzyme-Substrate Complex
Michealis-Menten Kinetics
Inhibition of Enzyme Reactions
(Chapter 7 Fogler, page 409-416)
Competitive Inhibition
Uncompetitive Inhibition
Non-competitive Inhibition
Types of Bioreactors (Lecture notes/Assignment)
Rates & Kinetics of Biological Processes
(Lecture notes)
 Enzymatic Reaction Fundamental
(Enzyme-Substrate Complex)
Enzymes are proteins that act as biological catalysts.
Enzymes provide a pathway for the substrate to
proceed at a faster rate. The substrate, S, reacts to
form product P:
Slow
S P
Enzyme-Substrate
Complex
(an active Fast ES
intermediate)

Since enzyme are indeed a type of catalyst, they have

characters similar to catalyst:
usually present in small quantity
not consumed during the course of reaction
do not effect the chemical reaction equilibrium
A given enzyme can only catalyze one type of reaction.
since they are specific, unwanted products are
Examples of
enzymes

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 How do substrate and enzyme interact??

There are 2 models to describe substrate-

enzyme interactions:
a) Lock-and-key model

b) Induced fit model

 Enzyme Kinetics
(Michaelis-Menten Kinetics)
Kinetics is the study of the rated of chemical
reactions.
The rates of biological reactions are greatly increased
by enzyme catalysts.
Enzyme kinetics is based upon the elementary
kinetics.
The mechanism of enzymes reactions are similar as
catalytic reactions. Substrate (S) adsorb reversibly on
the Enzyme (E), and then reacts to form product (P)
and leaves a free enzyme site for further reaction.
The rate of reaction catalyzed by enzyme can usually
be described by the Michaelis-Menten kinetics.
 Enzyme Kinetics
(Michaelis-Menten Kinetics)

cont.
To further illustrate enzyme kinetics, we
will use
the example of urea (NH2CONH2) decomposition by
the enzyme urease:
NH2CONH2 + UreaseH O 2 2NH3 + CO2 +
Urease
H2O
S + E P
+ E
k
The corresponding mechanism is:
1
r1 = k1(E)
E+S
k 2
ES (S)
r2 = k2(ES)
ES k E+S
3
r3 = k3(ES)
ES + W P+E (W)
Activity:Answer:
write the-rnet rate disappearance of7-19
s = -d(S)/dt = k1(E)(S) Eq.
S. Fogler
 Enzyme Kinetics
(Michaelis-Menten Kinetics)

cont.
From previous slide, we see that the net rate
disappearance
-rs of
= S-d(S)/dt
is: = k1(E)(S) k2 (ES)
Eq. 7-19 Fogler

However, (ES) is immeasurable. Why??

Therefore, as usual, we will substitute (ES) into the

form of measurable quantity. This is done by assuming
rES = d(E S)/dt = 0
PSSH on (ES):

k 1 [E ][S ]
rES = k1(E)(S) k
[E S ] 2 (ES) k3 (ES)(W) =0
Eq. 7-21 Fogler
k 2 k 3 [W ]

 Enzyme Kinetics
(Michaelis-Menten Kinetics)
cont. after we substitute (ES) found
However, in Eq.
7-21 into Eq. 7-19, Eq. 7-19 still cannot be used.

Good news is, we can measure the total

concentration of the enzyme in the system, (Et):
(Et) = (E) +
Eq. 7-23 Fogler
(ES)
By substituting Eqs. 7-21 and 7-23 into Eq. 7-
19, finally the measurable net rate of
disappearance of S is: k 1k 3[ W][S][Et ]
-rS = Eq. 7-24 Fogler
k 1[S] + k 2 + k 3 [ W ]
 Enzyme Kinetics
(Michaelis-Menten Kinetics)
cont.
k 1k 3 [ W ][S][Et ]
-rS = Eq. 7-24 Fogler
k 1[S] + k 2 + k 3 [ W ]

k cat = k 3[ W]
k cat + k 2
KM =
k1
Vmax = k cat [Et ]

k cat [Et ][S] Vmax [S] (Michaelis-Menten

-rS = =
[S] + K M K M + [S] equation)
Eq. 7-25 & Eq. 7-26
Fogler

Class Activity: Interpret the meaning or

significance of kcat, KM and Vmax
Enzyme Kinetics (Michaelis-Menten
Kinetics) cont.
Vmax [S] (Michaelis-Menten
-rS = Eq. 7-26 Fogler
if, K M + [S] equation)
if,
if,

1 K 1 1
M
r s Vmax [S ] Vmax
-rs or rp

Michaelis-Menten plot Lineweaver-Burk plot

reaction rate versus substrate
 Inhibition of Enzyme Reactions
Temperature and pH greatly influence the rates of
enzyme-catalyzed reactions.
However, there is also another factor that greatly
the presence ofreactions:
influence the rates of enzyme-catalyzed inhibitors.
Inhibitor is defined as species that interacts with
enzymes and render the enzyme ineffective to catalyze
its specific reaction.

www.vwmin.org
 Inhibition of Enzyme Reactions (cont.)
However, not all consequences of inhibitors are bad. There are
also beneficial inhibitors, such as one used in leukemia
treatment.
In other words, inhibitors are actually a way of controlling and
regulating the enzymatic activity.
2 categories of enzymatic
inhibition

Irreversible Reversible
inhibition inhibition

Inhibitor binds to Inhibitors can

enzyme very tightly. dissociate from
It binds so strongly enzyme under certain
that it is very unlikely conditions (certain
that the inhibitor environment)
Competiti Noncompeti
Competiti 3 types of
will ever dissociate
ve reversible tive
from the enzyme
Inhibition inhibition Inhibition
Uncompetit
ive
Inhibition of Enzyme Reactions (cont.)
Competitive Inhibition
 Fogler (4th Ed.)-Elements of Chemical Reaction Engineering, Copyright 2006

Inhibition of Enzyme Reactions (cont.) Pearson Education, Inc.

Competitive Inhibition (cont.)

Inhibition of Enzyme Reactions (cont.)
Uncompetitive Inhibition
Inhibition of Enzyme Reactions (cont.)
Uncompetitive Inhibition (cont.)

Fogler (4th Ed.)-Elements of Chemical Reaction Engineering, Copyright 2006

Inhibition of Enzyme Reactions (cont.)
Noncompetitive Inhibition
 Inhibition of Enzyme Reactions (cont.)
Noncompetitive Inhibition (cont.)

Fogler (4th Ed.)-Elements of Chemical Reaction Engineering, Copyright 2006

 Inhibition of Enzyme Reactions (cont.)
Summary of Lineweaver-Burk plots

Fogler (4th Ed.)-Elements of Chemical Reaction Engineering, Copyright 2006

 Bioreactors

Glacial Lakes Energy in Watertown, South Dakota

47+ million gallon per year ethanol production .

World's Largest Industrial Fermenter (Chem. Eng. News,10-Apr-78)

The fermenter is 200' high and 25 ft diam.

 What is a bioreactor?
Bioreactor: device, usually a vessel, used to direct the activity of a
biological catalyst to achieve a desired chemical transformation.

Fermenter: a type of bioreactor

in which the biocatalyst is a
living cell.

Pre-filtration

Input

Nutrients tank
Waste

Recycle

Product
Bioreactor
Product
separation & purification
 Challenges in Bioreactor Design

1. Aerobic bioreactor: Need adequate

mixing and aeration
2. Anaerobic bioreactor: no need for
sparging or agitation
 Bioreactor Operation Modes
-1. Batch Operation
A foam breaker may be installed to disperse foam
A batch
bioreactor is
normally
equipped with dC s rmax CS
an agitator to r
dt K m CS
mix the
reactant, and
the pH of the
reactant is Batch
operation
maintained by with stirring
employing
Cs 0
either buffer
Change of
K m ln Cs 0 Cs rmax t
solution Cs
or with
a pH Cs
controllertime, t
 Bioreactor Operation Modes
-2. Plug-flow mode
An ideal plug-flow reactor
can approximate the long
In a plug-flow tube, packed-bed and
reactor, the hollow fiber or multistaged
substrate reactor
enters one end F, Cs0 V F, Cs
of a cylindrical
tube with is V Residenc
packed with t=0 e time
immobilized
F
Continuous
enzyme and the operation without
product steam stirring
leaves at the
Cs 0
other end. K m ln Cs 0 Cs rmax t
Cs
 Bioreactor Operation Modes
-3. Continuous stirred-tank
A continuous
F, Cs0
stirred-tank
reactor (CSTR)
is an ideal
F, Cs
reactor which is V
based on the
assumption that
the reactants Continuous
are well mixed. operation
with stirring
 Bioreactor Operation Modes
-3. Continuous stirred-tank reactor-Con.
Mass balance of
F, Cs0
substrate:
Input - Output Consumption Accumulation

F, Cs
V dC s
FCs 0 FCs rsV V
dt
dC s
Steady 0
dt
state:
Michaelis- rmax C S
Menten r
K m CS
rate:
rmax Cs
FC s 0 FC s V 0
K m Cs
 Bioreactor Operation Modes
-3. Continuous stirred-tank reactor-Con.
Mass balance of
F, Cs0
substrate:
rmax Cs
FCs 0 FCs V 0
F, Cs K m Cs
V

F rmax Cs


V Cs 0 Cs K m Cs
F 1

V
rmax Cs
Cs K m
Cs 0 Cs
Rates & Kinetics of Biological
Processes

A ns S n A A n P P
Abiological species i.e organism or
cells
Ssubstrate or food supply
Pmetabolic product

nA>1 organism growth

(reproduction)
 Kinetics of Reproduction

A S 2A P r kC ACS
If excess
nutrient CS CSO

r kC ACSO
In batch reactor,
dC A
kC A CSO
dt
C A C Ao e kC so t
 Metabolism

A is not reproducing, therefore nA=1

A S A P r kC ACS

SP

C A C AO
CA is constant,
In batch reactor,
dCS
kC Ao CS
dt
kC Ao t
C S C So e
CSO CS C P Material balance

kC Ao t
C P C So (1 e )
 References
Schmidt, L.D. (2005). The Engineering of Chemical Reactions,
2nd edition, New York: Oxford University Press.
Fogler, H.S. (2006). Elements of Chemical Reaction
Engineering, 4th Edition, New Jersey: Prentice Hall.
Levenspiel, O. (1999). Chemical Reaction Engineering, 3rd
Edition, New York: John Wiley.
Pn. Hasyimi Rahmat, FKK UiTM Shah Alam.
Pn. Sharmeela Matali, FKK UiTM Shah Alam.
Pn. Shafiza Hashib, FKK UiTM Shah Alam.
Pn. Siti Wahidah Puasa, FKK UiTM Shah Alam.

FACULTY OF CHEMICAL
CPE624/CHE625
ENGINEERING