A + M → A∗ + M
• The idea was suggested in 1922 by F.A.Lindermann , acitve
intermediates were experimentally observed using
femtosecond spectroscopy by A.Zewail (Nobel Prize 1999)
cyclobutane
x2 ethylene
Pseudo –Steady-State Hypothesis (PSSH)
• Decomposition of the intermediate doesn’t occur
instantaneously, activated species have finite life time
• Active intermediates react as fast as they are formed, so their
net rate of formation is zero:
N
rA* = ∑ riA* ≡ 0
i =1
Pseudo –Steady-State Hypothesis (PSSH)
• Gas-phase decomposition of azomethane into
ethane and N2:
( CH 3 )2 N 2 → C2 H 6 + N 2
• experimentally found to follow
– 1st order at pressures above 1atm rC2 H 6 ∝ C AZO
2
– 2nd order below 50mmHg rC2 H 6 ∝ C AZO
Pseudo –Steady-State Hypothesis (PSSH)
• Suggested mechanism:
( 3 )2 2 ( 3 )2 2
CH N + CH N k1 AZO*
→ ( CH 3 )2 N 2 + ( CH 3 )2 N 2 *
( CH 3 )2 N 2 * + ( CH 3 )2 N 2 k2 AZO*
→ ( CH 3 )2 N 2 + ( CH 3 )2 N 2
( CH 3 )2 N 2 *
k3 AZO*
→ C2 H 6 + N 2
• the rate laws
r1 AZO* = k1 AZO*C 2 AZO
r2 AZO* = −k2 AZO*C AZO*C AZO
r3 AZO* = −k3 AZO*C AZO*
rAZO* = r1 AZO* + r2 AZO* + r3 AZO* ≡ 0
Pseudo –Steady-State Hypothesis (PSSH)
• solving for Cazo* and finding the rate of formation of product:
k1k3C 2 AZO
rCr H6 =
k2C AZO + k3
• at low concentrations
• at high concentrations
k1k3
k2C AZO k3 rCr H 6 = C AZO = kC AZO
k2
Pseudo –Steady-State Hypothesis (PSSH)
• Experimentally finding the mechanism:
k1C 2 AZO
rCr H 6 =
k ′C AZO + 1
Vmax ( S )
− rS =
( S ) + KM
Evaluation of Michaelis-Menten parameters
1 1 K 1
− = + M
rS Vmax Vmax ( S )
Vmax ( S )
−rS =
( S ) + KM
(S ) = KM 1
− + (S )
rS Vmax Vmax −r
−rS = Vmax − K M S
(S )
KM 1 Curea 0 X
Curea = Curea 0 (1 − X ) t= ln +
Vmax 1 − X Vmax
Briggs-Haldane equation
• If the reaction of forming the product from the enzyme-
substrate complex is reversible
E + S E iS E + P
temperature inactivation
due to enzyme denaturation
optimum
Nutritions required:
• Carbon source stationary phase: lack of some
• Nitrogen source nutriotients limits cell growth;
• Oxygen source many important products (e.g.
• Phosphate source antibiotics) produced at this phase.
• ….
• In many systems the product can inhibit cells growth (e.g. wine
making)
n
µmax Cs Cc Cp
rg = kobs kobs = 1 − *
K s + Cs C
p product concentration
where metabolism ceases
rd = ( kd + kt Ct ) Cc
death due to toxic environment
• Temperature effect: similar curve with max
Bioreactors: Stoichiometry
Cells
Substrate
→ More Cells + Product
• Yield coefficient for cells and substrate
Mass of new cells formed ∆CC
Yc / s = =−
Mass of substrate consumed ∆CS
µmax Cs Cc
rp = Yp / c
K s + Cs
Bioreactors: Stoichiometry
Cells
Substrate
→ More Cells + Product
• Yield coefficient for product in the stationary phase
Mass of product formed ∆CP
Yp / s = =−
Mass of substrate consumed ∆CS Usually secondary
nutritient
• Maintenance utilization term, typically m=0.05 h-1.
rsm = mCc
cells
→ Yc′/ s C + Yp′/ s P + maintance
S
− rs = Yc′/ s rg + Yp′/ s rp + mCc
Bioreactors: Stoichiometry
cells
S → Yc′/ s C + Yp′/ s P + maintance
− rs = Yc′/ s rg + Yp′/ s rp + mCc
• In the exponential phase we cannot separate substrate
consumption for cell growth and product formation
− rs = Ys / c rg + mCc
rp = rg Yp / c
• In stationary phase:
k p Csn
rp =
K sn + Csn
Ysn / p k p Csn
− rsn = Ysn / p rp + mCc = + mCc
K sn + Csn
Chemostat (CSTR) operation
• Mass balance:
dCc
– Cell balance V = v0Cco − vCc + (rg − rd )V
dt
dCs
– Substrate balance V = v0Cso − vCs + rsV
dt
Batch operation
• Mass balance: dCc
– Cell balance V = (rg − rd )V
dt
– Substrate
balance
growth phase stationary phase
dCs dCs
= Ys / c (−rg ) − mCc = Ys / p (−rp ) − mCc
dt dt
– Product balance
stationary phase
growth phase
dC p dC p
= rp = Yp / c rg = rp = Yp / s (−rs )
dt dt
Example 7-6
• Fermentation of Saccharomyces cerevisae in a batch reactor.
Plot the concentration of cells, substrate, the product and
growth rate as a function of time.
Initial cell concentration 1g/dm3, glucose 250g/dm3
C * p = 93 g / cm3 Yc / s = 0.08 g / g
n = 0.52 Yp / s = 0.45 g / g
µmax = 0.33 h −1 Yp / c = 5.6 g / g
K s = 1.7 g / dm3 kd = 0.01 h −1
m = 0.03 ( g substrate ) / ( g cell ⋅ h )
Physiologically based pharmacokinetics (PBPK)
• Chemical reaction engineering approach can be applied to
pharmacokinetics