Last Time 3/4/08

Increase (I) concentration increase slope intercept remains unchanged.

r
S
=
V
max
S
( )
K
M
1+
I
( )
K
I
|
\

|
.
|
+ S
( )


Uncompetitive Inhibition
Inhibition only has affinity for enzyme-substrate complex




Developing the rate law


(1)

(2)


E+S
k
1

k
2

E-S
k
cat
P
I + E-S
k
4

k
5

I-E-S

r
P
=r
S
=k
cat
E-S
( )

r
E-S
=0 =k
1
E
( )
S
( )
k
2
E-S
( )
k
cat
E-S
( )
k
4
I
( )
E-S
( )
+k
5
I-E-S
( )

r
I-E-S
= 0 =

+k
4
I
( )
E-S
( )
k
5
I-E-S
( )
Adding (1) and (2)

k
1
E
( )
S
( )
k
2
E-S
( )
k
cat
E-S
( )
= 0
E-S
( )
=
k
1
E
( )
S
( )
k
2
+k
cat
=
E
( )
S
( )
K
M

I-E-S
( )
=
k
4
k
5
I
( )
E-S
( )
=
I
( )
E-S
( )
K
I
=
I
( )
E
( )
S
( )
K
I
K
M
K
I
=
k
5
k
4
r
p
= k
cat
E-S
( )
=
k
cat
E
( )
S
( )
K
M
From (2)
Total enzyme

E
t
= E
( )
+ E-S
( )
+ I-E-S
( )
= E
( )
1+
S
( )
K
M
+
I
( )
S
( )
K
I
K
M
|
\

|
.
|
r
p
=
k
cat
E
t
S
( )
K
M
1+
S
( )
K
M
+
I
( )
S
( )
K
I
K
M
|
\

|
.
|

r
S
= r
P
=
V
max
S
( )
K
M
+ S
( )
1+
I
( )
K
I
|
\

|
.
|

r
S
= r
P
=
V
max
S
( )
K
M
+ S
( )
1+
I
( )
K
I
|
\

|
.
|

1
r
S
=
1
V
max
S
( )
K
M
+ S
( )
1+
I
( )
K
I
|
\

|
.
|
|
\

|
.
|
=
K
M
V
max
1
S
( )
|
\

|
.
|
+
1
V
max
1+
I
( )
K
I
|
\

|
.
|
Slope remains the same but intercept changes as inhibitor concentration is increased


Lecture16 Thursday 3/6/08
Enzyme Kinetics
Noncompetive Inhibition
Bioreactors
Monod Equation
Yield Coeficients
Washout


Bioreactors




D=v
o
/ V

Noncompetitive, Bioreactors
More uncompetitive inhibition
|
|
.
|

\
|
+
|
|
.
|

\
|
+
=

|
|
.
|

\
|
+ +
= =
S
m
I
I
S
I
I
S m
S
S P
C V
k
V
k
C
r
k
C
C k
C V
r r
1
1
1
1
max max
max
Only intercept
changes
S
C
1
S
r
1

Increasing I
No I
Lineweaver-Burk
Noncompetitive inhibition (Mixed)
Both slope and
intercept change
S
C
1
S
r
1

Increasing I
No I
E + S ES P

EI + S ESI
+I +I -I -I
( )
|
|
.
|

\
|
+
|
|
.
|

\
|
+
|
|
.
|

\
|
+ =

|
|
.
|

\
|
+ +
=
I
I
S
m
I
I
S
I
I
S m
S
S
k
C
C V
k
k
C
V r
k
C
C k
C V
r
1
1
1
1 1
1
max max
max
Bioreactors
Cells + substrate product + more cells
Cell growth can be modeled with enzyme
equations since all cell reactors are enzyme
catalyzed
Lag
phase
Exponential
growth phase
stationary
phase
Death
phase
Algorithm
1) Mass balance (later)
2) Rates
( )
=
|
|
.
|

\
|
=
+
=
+
= =
*
*
max
max
1
equation) monad (
P
n
P
P
OBS
OBS C
S S
S
g
S S
S
C g
C
C
C
k
k C
C k
C
r
C k
C
C r

sometimes
Product concentration at which all metabolism
ceases (93 g/L for wine production, n = )
Substrate is used to form both products and cell maintenance
(rate of substrate disappearance for maintenance C sm
mC r =
3) Stoichiometry yield coefficients
C D D
C g C S S g C P P
C P
S C
C S S C
C k r
mC r y r r y r
y
y
y y
=
+ = =
=
= =
/ /
/
/
/ /


formed cells new of mass
formed product of mass
1

consumed substrate mass
formed cells of mass
1) Mass balance







Parameters:
( )
( )
( )
(product)
) (substrate
rate) (dilution
1

(cells) 0
0
0 0 0
0
0
P P
P
S S S
S
S S S
S
d g C
C
d g C
C
r DC
dt
dC
r C C D
dt
dC
V r C v C v
dt
dC
V
V
v
D r r DC
dt
dC
V r r C v
dt
dC
V
=
+ =
+ =
= = + =
+ =
t
2) Rates
C D D
C g C S S
g C P P
n
P
P
OBS
OBS C
S S
S
g
C k r
mC r y r
r y r
C
C
K
K C
C k
C
r
=
=
=
|
|
.
|

\
|
=
+
=
/
/
*
max
1

V DV v C v m C v m
C D m y y k k n C
P p C
S C P C S D S P
, , ,
, , , , , , , , ,
0 0 0
0 / /
*
max
= = =