Enzyme Kinetics

CHE182
Enzyme’s Lock and key theory
 Enzyme’s Reaction Mechanism

• S + E ↔ ES Rapid/in equilibrium

• ES → P + E slow

• Apply steady state assumption (SSA)

• Apply rate determining state theory (RDS)
 Michaelis-Menten Equation

• Where v is the velocity or rate of product formation

• vmax is maximum rate of product formation.
• [S] is the substrate concentration.
• KM is the Michaelis Menten Constant.
 Sample Problem 1

In an experiment conducted to evaluate the Michaelis-

Menten constant, it was found that 1 g of bacteria could
decompose the waste at a maximum rate of 35 g/day
when the waste concentration was high. It was also found
that the same quantity of bacteria would decompose
waste at a rate of 18 g/day when the waste concentration
was 20 mg/L. Calculate rate of waste decomposition by 2
g of bacteria if the waste concentration were maintained
at 8 mg/L.
A. 10.4 g/day B. 15.6 g/day C. 20.8 g/day D. 31.2 g/day
 Quiz 5

1. _______ tags protein for destruction. 2pts

2. What vitamin helps in deamination of serine? 2 pts
3. What amino acid transport ammonia from muscle?
4. Ornithine and Carbamoyl phosphate condensed to
form _______. 2 pts
5. The second ammonia of Urea is coming from this
amono acid. 2 pts
Lineweaver-Burk plot

y = mx + b
Sample Problem 2
Hanes-Woolf plot

y= b + mx
Eadie-Hofstee Plot equation.

y= b + mx
MM and Batch Reactors

=
MM and Batch Reactors

+
 Sample Problem

• The initial concentration of glucose in a batch

reactor fermentation media is 100 g/L. Previous rate
study shows, at the same enzyme concentration, that
the maximum rate of conversion of glucose to
ethanol is 3 g/L.h and the Michaelis-Menten
constant is 2 g/L. Calculate the number of hours it
will take the yeast to reduce the glucose
concentration to 10 g/L.
 CSTR and MM

• Fso – Fs = -rs V

• Where Fso is the initial substrate molar flow rate (moles of

solute/time)

• V is the volume of the reactor

• -rs is the Michaelis-Menten reaction rate

 CSTR and MM

• τ = V/v

• Tau is residence time.

• Reactor volume/ volumetric flow rate.

CSTR and MM
 CSTR and MM

• Consider a 10 L CSTR maintained at 310 K where

the initial glucose concentration is 1.00 moles/L
flowing at 1.00 L/s. Previous study shows that Vmax
is 1.2 moles/L.s and KM is 0.4 mol /L. Calculate the
final glucose concentration.
A continuously stirred tank reactor (CSTR) with a working
volume of 15 m3 is used for the production of baker's yeast.
The overall reaction for aerobic fermentation is expressed as:
0.556 C6H12O6 + 0.301 NH3 + 1.43 02 → 0.301C6H10O3N +
2.283 H2O + 1.532 CO
The empirical formula for the biomass is C6H10O3N (144) and
molasses contains 40.8% w/v glucose (180).Calculate the daily
molasses consumption to maintain a daily production of one
metric ton if the residual sugar in the outgoing broth is 1.40 %
w/v glucose.

A. 1465L/day B. 2930 L/day

C. 4395 L/day D. 5860L/day
 Specific Growth Rate, u

• u = 1/τ
• u = v/V
• The unit is per unit time.
 Sample Problem

2. The specific growth rate of the yeast

is____
A. 0.13/day
B. 0.26/day
C. 0.39/day
D. 0.52/day
 Sample Problem

• For a given species of a microorganism that doubles

every 3 h, what is the mass of biomass that may be
expected from 100 liters of seed if each liter contains
8 grams biomass and the fermentation culture was
maintained for 24 h is____________
• A. 102 kg B. 204 kg
• C. 306 kg D. 408 kg
 Quiz

• An aqueous solution of molasses contains 15% by

weight sucrose (C12H22O11). The CO2 formed in the
reaction can be considered as having a negligible
solubility in the solution. Determine the % wt
ethanol in solution after 95% of the sucrose has been
converted to ethyl alcohol by fermentation.
• A. 2.07% B. 4.14%
• C. 7.67% D. 8.27%
 Old board problem
• Albizzia folcataria, a specie of plywood, after an initial
thermochemical hydrolysis yielded 25 % maltose, 3% sucrose, 12
% cellobiose, 43 % oligosaccharides and 17%non carbohydrates
residues. The resulting hydrolyzate is passed through a column of
immobilized enzyme systems so that all types of dissaccharides
are converted further to hexose units. In alcohol fermentation,
the rule of thumb is 10% of the substrate is converted to
biomass and 90% to alcohol. What mass of ethanol is expected
to be produced per six-day week if one metric ton of pulpwood
is processed daily?
• A. 1162kg B. 1223 kg
• C. 1885 kg D. 2526 kg
 Quiz 2/Old board problem

• One metric ton of wild potatoes were hydrolyzed

enzymatically and the hydrolyzate contained the following:
xylose 2% glucose 16% oligosaccharides 36% pentose 3%
maltose 20% others 23%.
It was observed that of the hydrolyzate, only maltose and
glucose were fermentable by alcohol yeast.
Calculate the mass of alcohol that can theoretically be
expected from the hydrolyzate.
• a. 189kg b. 95kg c. 338 kg d. 371kg
Plug Flow Reactor and MM
Plug Flow Reactor and MM
MM and Plug Flow Reactors

+
 Sample Problem

• The initial concentration of glucose in a plug flow

reactor fermentation media is 100 g/L. Previous rate
study shows, at the same enzyme concentration, that
the maximum rate of conversion of glucose to
ethanol is 3 g/L.h and the Michaelis-Menten
constant is 2 g/L. Calculate the length of the plug
flow reactor needed if the diameter of the pipe use is
0.03 m to reduce the glucose concentration to 10 g/L
if the flow rate is 1000 L per day.
Competitive Inhibition
 Competitive Inhibition

E + S → ES K1
E + I → EI
Therefore K1 decreases!

KM = (K2 + K3)/K1

Thus KM increases!
 Competitive Inhibition

Vmax = K3 [E]t
K3 and [E]t are constant !
Therefore Vmax don’t change.
Non-Competitive Inhibition
 Non-competitive Inhibition

ES + I → ESI
Vmax = K3 [E]t
K3 decreases !
Therefore Vmax decreases.
 Non-Competitive Inhibition

E + I → EI
E + S → ES
Therefore K1 decreases!
And since K3 also decreases
KM is unchanged!
Uncompetitive Inhibition
 Uncompetitive Inhibition

ES + I → ESI
ES → E + P
Therefore K3 decreases
While K1 and K2 remain constant.
Therefore Vmax and KM decreases.

KM = (K2 + K3)/K1 Vmax = K3 [E]t

Eisenthal and Cornish-Bowden Plot
The kinetics of an enzyme is measured in the presence
and absence of a 2 mM Inhibitor (I)

a. What are the values of Vmax and Km in the absence

of Inhibitor (I)? In its presence?
b. What type of inhibition is it?
 The enzyme kinetics in the last problem is studied in the
presence of another Inhibitor. The concentration of this
inhibitor is 100 uM.

a. What are the values of Vmax and Km in the presence of

Inhibitor (I)?
b. What type of inhibition is it?