10.

37 Chemical and Biological Reaction Engineering, Spring 2007

Prof. K. Dane Wittrup
Lecture 3: Kinetics of Cell Growth and Enzymes

This lecture covers: cell growth kinetics, substrate uptake and product formation in
microbial growth, enzyme kinetics, and the Michaelis-Menten rate form.

Biological Rate Laws- Enzymes and Cell Growth

Rate Law: − rA = f (C A ,CB ,CP ,T , pH ,...)

Why would you need a rate law?

-Predictive description of a production process
-Design tool for forming a desired product
-Consistency or inconsistency with alternative mechanism

For a hypothesized mechanism, we can often derive an exact, closed form analytical
solution.
If not, it’s used to approximate:
-some reactions go rapidly to equilibrium
-the concentrations of some species rapidly reach their steady state values
-the rate-limiting step
Or, as a last resort- numerical solution

X=first order − rA = kC A

-rA =second order −rA = kC A 2

X
=zero order − rA = k
X
X
X
X

CA

Figure 1. Rate versus concentration graphs for zero, first, and second order
reactions.

Enzymes-Biological Catalysts
S=Substrate
E=Enzyme

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 Activation energy
decreases with E
(enzyme)
Energy

P
Reaction Progress
Figure 2. An energy diagram for a reaction with and without an enzyme. The
activation energy is lower with an enzyme, so the reaction proceeds faster.

CS Initial Rate
dCS
− rS = −
dt t =0

Time
Figure 3. The slope of the concentration versus time curve at time = 0 can give the
initial rate.

Michaelis Menten
~1st order
~0th order
-rS
X X X X
X
X
X
X What gives change
X from 1st order to 0th
X order?
X
CS

Figure 4. Michaelis-Menten kinetics.

Hypothesized Mechanism:
-Encounter complex ES formed
-Irreversible reaction occurs In more complex cases,
-Rapid release of product from complex these are not always true
-Assume rapid equilibrium is reached in the formation of ES

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Later, Briggs-Haldane derived a law with steady state assumption for ES and they
got the same rate law as Michaelis-Menten. However, the Briggs-Haldane method is
more generally applicable.

ZZZ
E + S YZZ1
X
Z ES
k
k −1

ES ⎯⎯→ E + P
kcat

dCES
Steady state assumption on ES → =0
dt

Material Balance on ES:

dCES
= k1CE CS − k−1CES − kcat CES ≈ 0 (steady state)
dt
Free enzyme concentration ≠ CE ,0
because CE ,0  CS ,0
CE ,0 = CE + CES
0 = k1 (CE ,0 − CES )CS − CES (k−1 + kcat )

Cs ≈ CS ,0

Solve for CES (steady state solution)

CE ,0CS
CES =
k−1 + kcat
+ CS
k1
dC p kcat CE ,0CS V C
− rS = rp = = kcat CES = = max S
k−1 + kcat
dt + C S K m + CS
k1
Vmax = kcat CE ,0 = maximum reaction velocity
k−1 + kcat
Km = = Michaelis constant
k1
This model fits the data:
Vmax
-As CS  K m , − rS → CS
Km
-As CS  K m , − rS → Vmax

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 -rS
X X X
X The rate flattens out
X
because all of the
X
X enzyme is bound.
X
X
X
CS

Figure 5. Michaelis-Menten kinetics.

-This rate law reappears in heterogeneous catalysis (Langmuir-Hinshelwood)

-In the actual Michaelis-Menten derivation Kmax was an equilibrium constant
-Briggs and Haldane showed that this rate law still works for steady state
-Steady state approximation is more general
-Steady state approximation is consistent with the data
-Steady state is defined over a limited period of time. Because there is an
irreversible step, eventually all of the intermediate will be consumed.
-Equilibrium assumption is not exactly correct. The irreversible step prevents
true equilibrium.

Growth Kinetics
A population of single cells, growing without limitations
N=#cells per volume
dN
= μ N , μ = constant, specific growth rate
dt
N = N 0e μ t − exponential growth while true, "Malthusian growth"

Growth Stops
-Run out of nutrients
-Accumulate toxic byproducts

Nutrient Effect on μ (Monod)

μ C
μ = max S
K S + CS
This expression is purely empirical (no mechanistic meaning). Even the fit is not that
good.

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dN μ C
= max S N
dt K S + CS
We will return to this when we
dCS −1 look at bioreactors
= μN
dt Yx s

I=inhibitor or toxin

μ μ ∝ e − kC I

1
μ∝ All forms fit the data
(1 − kCI )
kI
μ∝
k I + CI

CI

Figure 6. Growth rate versus concentration of inhibitor. Many functional forms fit
the curve.

CS=concentration of growth limiting

μ
substrate, often glucose
X X X
X
X
X
X
X
X KS
X
CS

Figure 7. Growth rate versus concentration of growth limiting substrate.

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Cell Growth as a Chemical Reaction
Aerobic growth:
Carbon source + O2 + Nitrogen Source → Biomass + Byproducts + H2O +CO2

CH1.8O0.5N0.2
(for a typical microbial culture)

A yield coefficient can be defined:

ΔA
YA B =
ΔB
A=biomass, byproducts, CO2, heat (any of these)
B=carbon source or oxygen

g biomass
For glucose, Yx s ≈ 0.6 ± 0.1
g glucose
g biomass
For oxygen, Yx O2 ≈ 1.9 ± 0.7
g O2

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