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12/3/2015

Chapter 3
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)

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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:
S Slow  P
Enzyme-Substrate Complex
(an active intermediate)

ES
Fast

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 easily controlled in
enzyme-catalyzed reactions.

Examples of enzymes

http://www.google.com/patents/EP1706130A2?cl=en

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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.

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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 + Urease H2O 2NH3 + CO2 + Urease
H2O
S
+
E
P
+
E
The corresponding mechanism is:
k1
r1 = k1(E)(S)
E+S
ES
r2 = k2(ES)
ES k2
E+S
r3 = k3(ES)(W)
ES + W k3
P+E
Activity: write the net rate disappearance of S.

Answer: -rs = -d(S)/dt = k1(E)(S) k2 (ES)

Eq. 7-19 Fogler

Enzyme Kinetics
(Michaelis-Menten Kinetics) cont.
From previous slide, we see that the net rate disappearance of S is:
-rs = -d(S)/dt = k1(E)(S) k2 (ES)
However, (ES) is immeasurable. Why??

Eq. 7-19 Fogler

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


measurable quantity. This is done by assuming PSSH on (ES):

rES = d(E S)/dt = 0


rES = k1(E)(S) k2 (ES) k3 (ES)(W) = 0

[E S ] =

k 1 [E ][S ]
k 2 + k 3 [W ]

Eq. 7-21 Fogler

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Enzyme Kinetics
(Michaelis-Menten Kinetics) cont.
However, after we substitute (ES) found 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) + (ES)

Eq. 7-23 Fogler

By substituting Eqs. 7-21 and 7-23 into Eq. 7-19, finally the
measurable net rate of disappearance of S is:
-rS =

k 1k 3[ W][S][Et ]
k 1[S] + k 2 + k 3 [ W ]

Eq. 7-24 Fogler

Enzyme Kinetics
(Michaelis-Menten Kinetics) cont.
k 1k 3 [ W][S][Et ]
-rS =
k 1[S] + k 2 + k 3 [ W ]

Eq. 7-24 Fogler

k cat = k 3 [ W ]
k
+ k2
K M = cat
k1
Vmax = k cat [Et ]

k [E ][S] Vmax [S]


=
-rS = cat t
K M + [S ]
[S] + K M

(Michaelis-Menten equation)
Eq. 7-25 & Eq. 7-26 Fogler

Class Activity: Interpret the meaning or significance of kcat, KM


and Vmax

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Enzyme Kinetics (Michaelis-Menten Kinetics) cont.


if,

-rS =

Vmax [S] (Michaelis-Menten equation)


K M + [S] Eq. 7-26 Fogler

if,
if,

-rs or rp

K
1
1
1
= M
+
r s Vmax [S ] Vmax

Michaelis-Menten plot reaction rate


versus substrate concentration

Lineweaver-Burk plot

Inhibition of Enzyme Reactions


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

www.vwmin.org

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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 inhibition

Reversible inhibition

Inhibitor binds to enzyme


very tightly. It binds so
strongly that it is very
unlikely that the inhibitor
will ever dissociate from
the enzyme

Inhibitors can dissociate


from enzyme under certain
conditions (certain
environment)

Competitive
Inhibition

3 types of
reversible inhibition

Noncompetitive
Inhibition

Uncompetitive
Inhibition

Inhibition of Enzyme Reactions (cont.)


Competitive Inhibition

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

Inhibition of Enzyme Reactions (cont.)


Competitive Inhibition (cont.)

Inhibition of Enzyme Reactions (cont.)


Uncompetitive Inhibition

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Inhibition of Enzyme Reactions (cont.)


Uncompetitive Inhibition (cont.)

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

Inhibition of Enzyme Reactions (cont.)


Noncompetitive Inhibition

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Inhibition of Enzyme Reactions (cont.)


Noncompetitive Inhibition (cont.)

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

Inhibition of Enzyme Reactions (cont.)


Summary of Lineweaver
Lineweaver--Burk plots

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

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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.

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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

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Bioreactor Operation Modes


-1. Batch Operation
A batch bioreactor is
normally equipped
with an agitator to
mix the reactant, and
the pH of the reactant
is maintained by
employing either
buffer solution or a pH
controller

A foam breaker may be installed to disperse foam

r=

dC s
r C
= max S
dt
K m + CS

Batch operation
with stirring

K m ln

Change of Cs
with time, t

Cs0
+ (C s 0 C s ) = rmax t
Cs

Bioreactor Operation Modes


-2. Plug-flow mode
In a plug-flow reactor,
the substrate enters
one end of a
cylindrical tube with is
packed with
immobilized enzyme
and the product
steam leaves at the
other end.

An ideal plug-flow reactor can


approximate the long tube,
packed-bed and hollow fiber or
multistaged reactor

F, Cs0

F, Cs

t=0

V
F

Residence
time

Continuous operation
without stirring

K m ln

Cs0
+ (C s 0 C s ) = rmax t
Cs

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Bioreactor Operation Modes


-3. Continuous stirred-tank
A continuous stirredtank reactor (CSTR) is
an ideal reactor which
is based on the
assumption that the
reactants are well
mixed.

F, Cs0

F, Cs
V

Continuous
operation with
stirring

Bioreactor Operation Modes


-3. Continuous stirred-tank reactor-Con.
Mass balance of substrate:

F, Cs0

Input - Output Consumptio n = Accumulation


F, Cs
V

FC s 0 FC s rsV = V
Steady state:

dC s
dt

dC s
=0
dt

Michaelisrmax C S
Menten rate: r = K + C
m
S

FC s 0 FC s V

rmax C s
=0
K m + Cs

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Bioreactor Operation Modes


-3. Continuous stirred-tank reactor-Con.
Mass balance of substrate:

F, Cs0

FC s 0 FC s + V
F, Cs

rmax C s
=0
K m + Cs

F
rmax C s
=
V (C s 0 C s ) K m + C s

Cs = K m +

rmax C s
Cs0 Cs

F 1
=
V

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
nA=1

organism growth (reproduction)


metabolism

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Kinetics of Reproduction

A + S 2A + P

r = kCACS

If excess nutrient CS = CSO

r = kCACSO

In batch reactor, dC
A
dt

= kC ACSO

C A = C Ao e kC so t

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Metabolism
A is not reproducing, therefore nA=1

A+ S A+ P

r = kCACS

S P

CA is constant, CA = CAO

In batch reactor,
dCS
= kC AoCS
dt

C S = C So e
CSO C S = C P

kC

Ao t

Material balance

C P = C So (1 e kC Ao t )

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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.

CPE624/CHE625

FACULTY OF CHEMICAL ENGINEERING

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