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= 1.65mumol/min
0.1 mg/ml x 0.1ml
= 1.65 mumol/min
1ml of enzyme
= 1.65 units/ml of enzyme
S vs t Graph
1.2
y = 0.033x + 0.0391
1
0.8
0.6
0.4
0.2
0
0
10
15
20
25
30
35
a) Michaelis-Menten approach
The rate of product formation.
d[p]
dt
( )
ubstitute
into
:
[(
(
Make
) (
)( )
)] ( )
)( )
)( )
as a subject:
(
)( )
)( )
)( )
)( )
[(
(
(
(
ub
into
)( )
)( )
)( )
)( )( )
)( )( )
as a subject,
(
)(
( )( ))
)
(
into
)( )( )
)( )( )
)(
)
(
ub
)( )
)
Make
)( )]
( )( ))
)( )( )
( )
( )( )
Since
)( )( )
)
( )
( )( )
)( )( )
)
( )
( )( )
)
( )( )
)
( )
( )( )
b) Since [ ]
[ ]
d[p]
dt
( )( )
( )( )
(ES)1
(ES)1 k3 (ES)2
(ES)2 k2 E+P
V=
[ ]
= k5 [ES] 2
Vm [S]
k2 k4 + k4 [S] + [S]
k1 k3
k3
d[p]
dt
( )
ubstitute
into
:
[(
Make
) (
)( )
)] ( )
)( )
)( )
as a subject:
(
)( )
)( )
)( )
)( )
[(
(
(
(
ub
into
)( )
)( )
)( )
)( )( )
)( )( )
as a subject,
(
)(
( )( ))
)
(
into
)( )( )
)( )( )
)(
)
(
ub
)( )
)
Make
)( )]
( )( ))
)( )( )
( )
( )( )
Since
)( )( )
)
( )
( )( )
)( )( )
)
( )
( )( )
)
( )( )
)
( )
( )( )
b) Briggs-Haldane approach
Make as a subject,
d(
( )( )
dt
d(
dt
ubstitute
into
)(
)(
)(
)( )
( )
( )
( )
( )
ubstitute
)( )
( )( )
)( )
( )(
( ))
( )
( )
( )(
( )
( )(
( )(
(
( ))
into
)( )
:
(
)(
)(
)(
( )
)( )
( )(
(
( ) ( )
( )
( ) ( )(
(
( )
( ))
)
( ))
( )
)( )
)(
)(
( )
( ) ( )
( ) ( )
(
ubstitute
)(
( ))
( ) ( )(
( )
)(
( )
( )
( ))
( ) ( )
( ) ( )
(
( ) ( )
( )
)(
( )
( ))
into
d(p)
dt
( ) ( )
(
( ) ( )
( )
)(
( )
( ))
v
d(p)
dt
v
(
( ) ( )
( ) ( )
( )
)(
( )
( ))
Km=0.0032
Kinetic Parameters
Vmax
Km
Langmuir
0.2750
0.0047
Lineweaver-Burk
0.0191
0.0057
Eadie-Hofstee
0.2645
0.0043
Non-Linear Regression 0.2000
0.0032
Type of Plot
Langmuir Plot
0.06
y = 3.3133x + 0.0191
0.05
0.04
0.03
0.02
0.01
0
0
0.002
0.004
0.006
0.008
0.01
y = 0.0172x + 3.6342
50
100
150
200
250
300
350
Eadie-Hofstee Plot
0.25
0.2
0.15
y = -0.0043x + 0.2645
0.1
0.05
0
0
10
15
20
25
30
35
40
0.002
0.004
0.006
0.008
0.01
Enzyme is preserved,
d( )
dt
(
Assumptions:
[ ]small,
neg igib e
( )( )
)(
)(
(
(
into
))( )
)
( )
(
(
)
)
( ) )
( )
( )
(
( )
d(p)
dt
(
(
( )
into
( )
(
( )( )
(
)
Substitute
v
( )( )
Substitute equation
(
()
( )
( )
)
)
( )
( )
]/
v s
( )
[ ]
[
]
[
= 60mol/m3.min
F = 0.0001m3/min
V = 0.0003m3
( FCSo - FCs ) (Km + Cs)
= Vmax CsV
2
FCSo Km + FCSo Cs - FKm Cs FCs = Vmax CsV
(0.0001 (300)(200) + 0.0001(300)Cs 0.001(200)Cs 0.001Cs2 = 100 (0.0003)Cs )
6 + 0.03Cs 0.02Cs 0.001Cs2 = 0.03Cs
0.0001Cs2 + 0.02Cs 6 = 0
Cs=165mol/m3
(Cso-Cs)/ln(Cso/Cs)
120
y = 6.3852x + 59.571
100
80
(Cso-Cs)/ln(Cso/Cs)
60
Linear ((CsoCs)/ln(Cso/Cs))
40
20
0
0
[ ]
[ ]
Km = 0.03mol/L
rmax = 13mol/L min 60 = 780mol/L hr
F=10L/Hr
Cs=10mol/L
F=10L/Hr
Cs=0.5mol/L
CSTR
a) V = ?
CSTR @ Stead State
FCs0 - FCs + rSv = 0
F (Cs0 - Cs ) = rpv
(
10 (10 0.5) =
V = 0.129 liter
+ (Cs0 - Cs )
0.03 ln
= rmax t
9.95899 = 780t
t = 0.0123hr
t = V/F = 0.0123
V = 0.0123 10
= 0.123liter
F=0.5L/min
Cs0=50g/L
1L
F=0.5L/min
Cs1=?g/L
1L
F=0.5L/min
Cs2=? g/L
)
s
(1)
(25-0.5Cs1)(10+ Cs1)=7Cs1
250+25Cs1-5Cs1-0.5Cs12=7Cs1
0.5Cs12-13Cs1-250=0
Cs1=38.86g/L
)
s
(1)
F=0.5L/min
Cs0=50g/L
2L
F=0.5L/min
Cs1=?g/L
)
s
(2)
(25-0.5Cs1)(10+ Cs1)=14Cs1
250+25Cs1-5Cs1-0.5Cs12=14Cs1
0.5Cs12-6Cs1-250=0
Cs1=29.15g/L
Since in the Cs in two reactor system is less than Cs in one reactor system, therefore two reactor
system is more efficient than one reactor system as it indicates more substrates have been
consumed to form products.
k1 [E] [S]
k2
][ ]
[ ]
[ ]
[E0] = [ES] {1 +
][ ]
][
[ ]
[ ]
+(
[ ]
(
[ ]
[
[ ]
)]
[ ]
(
[ ]
)]}
)]
V=
[ ]
b)
Given:
[ ]
[ ]
KSP =
KSP =
[ESP] =
[ ]
[ ]
[ ]
)]
[ ]
)]
KPS =
[ ][ ][ ]
[
]
KPS =
[ ][ ][ ]
[
]
[ ][ ][ ]
[EPS] =
[ ][ ][ ]
][ ]
]
][ ]
]
[ESP] = [EPS]
KS KSP = KP KPS
=
[ ]
=
=
[
[ ]
[ ]
[ ]
]
[ ]
[ ]
[ ]
]
]
[ ]
[ ]
c)
Ks=Kps
Kp=Ksp
[ESP]=[EPS]
[ ]
(
( )
[
[ ]
[ ][ ]
)[
[ ]
][ ]
]
[ ]
[ ]
[ ]
][ ]
[ ])
[ ](
[ ]
[ ]
[ ]
[ ])
[ ]
[ ]
[ ])
(
(
Compare with
[ ]
[ ]
Hence, Vmax = (
Km=
d)
[ ])
[ ]
(
[ ])
[ ]
[ ]
[ ]
[ ]
*
[
[ ]
[ ]
[ ]
[ ]
[ ]
[ ]
[ ]
[ ]
[ ]
n(
[ ]
)
[ ]
[ ]
+
[ ] [ ]]
[ ] [ ]
[ ]
[ ]
[ ]
n(
[ ]
)
[ ]
n([ ] [ ])
(
[ ]
[ ]
)
n ([
Y=
M=
[
X= (
] [ ]
C=
So we can plot a graph of
[ ]
)
]
n ([
vs (
] [ ]
---- 1
Enzyme balance:
CEo = CE + CES
---- 2
---- 3
---- 4
---- 5
---- 7
---- 6
---- 8
By rearranging Equation 5,
CE = (k2/k1) Cs CES
From Equation 2,
CEo = [(k2/k1)CE + 1] CES
CES = CEo /[( k2/k1)CS +1]
---- 9
By rearranging Equation 6,
CES = [(K6/K5)CI ] CEIS1
From Equation 3,
CEo = CEIS +CES + (k2/k1) Cs CES
= {CEIS1 + [1 + (k2/k1) Cs]( K6/K5)CI }CEIS1
= {1 + [1 + (K2/K1) Cs ]( K6/K5)CI } CEIS1
CEIS1 = CEo/ {1 + [1 + (k2/k1) Cs ]( K6/K5)CI }
By rearranging Equation 7,
CE = (K4/K3) CEI
By rearranging Equation 8,
CEI = K8/K7CS CEIS2
---- 10
---- 11
X=
X = -0.016
X = -0.005
In Line weaver Burk Plot and Langmuir Plot both indicates its a competitive inhibitor
Data :
Lineweaver
1/s
1/Vo
312.5
9.009009
204.0816 6.756757
161.2903 6.993007
125
6.024096
105.2632
5
Langmuir
s
s/Vo
0.0032 0.028829
0.0049 0.033108
0.0062 0.043357
0.008
0.048193
0.0095
0.0475
1/Vi
16.94915
14.08451
10.98901
9.009009
8
S/Vi
0.054237
0.069014
0.068132
0.072072
0.076
Lineweaver-Burk Plot
20
18
16
14
12
10
8
6
4
2
0
y = 0.0439x + 3.8266
1/Vo
1/Vi
y = 0.0172x + 3.6342
Linear (1/Vo)
Linear (1/Vi)
100
200
300
400
Langmuir Plot
0.09
0.08
y = 2.9883x + 0.0489
0.07
0.06
s/Vo
y = 3.3133x + 0.0191
0.05
S/Vi
0.04
Linear (s/Vo)
0.03
Linear (S/Vi)
0.02
0.01
0
0
0.002
0.004
0.006
0.008
0.01
ES + S
k5 E + P
(E0 ) = (E) +(ES) + (ESS)
(E) = (E0 ) (ES) (ESS) -------V=
=
(
( )
= k5 (ES) ------
)( )
)
K2 / k1 = (E)(S) / (ES)
K2/k1 (ES) = (E0)(S) (ES)(S)
(ES)((k2/k1) + (S)) = (E0)(S) (ES)(S)2 /
(ES)( (k2/k1) + (S)( ) ) =
(E0)(S) (ES)(S)2
(E0)(S)
3
V=
=
( )
Solution 2.17
V= 5L
Cso = 100 mmol/L
F = 1 L/hr
Cs = 10m mol/L
a) F (s0 FCs = rp V
1(100-10) = rp (5)
Rp = 18 m mol/ L.min
Solution 2.18
[SO]1 = 0.1 mol/L
[S0]2 = 0.3mol/L
[ ][ ]
[
]
[ ][
]
]
[ ][
[ ][
]
]
[
[ ][
[
]
]
[ ][ ]
[
]
V1 =
= k5[ES1]
= k6[ES2]
[ES1] =
[
[
]
(1+
(1+
]
(
)}
[ ]
[ ]
[S1] =
---
[ ][
[
]
[E0] = [E2] [
+ [ES2]
(1+
)+ [ES2]
[
(1+
)+ [ES2]
]
)+ 1]
]
[S2] = e
---
As [S1] increases, [ES1] also increases as in eq.3. [P1] also increases as in eq.1. This also occurs in
[S2]. As [S1] increases, [ES1] also increases as in eq.4. [P2] also increases as in eq.2
Solution 2.19
Data :
s
6.7
3.5
1.7
s/v
22.33333
14
10.625
Langmuir Plot
25
y = 2.3722x + 6.2429
20
15
10
5
0
0
1/Vm = 2.3722
Vmax = 0.4215 mumol/L.min
Km/Vm = 6.2429
Km = 6.2429(0.4215)
=2.63mumol/L
The dissociation constant for the first equilibrium reaction is the same as that of the third
equilibrium reaction.
The dissociation constant for the second equilibrium reaction is the same as that of the
fourth equilibrium reaction.
k
k2
K S 6 K IS
k1
k5
k
k4
K I 8 K SI
k3
k7
If the slower reaction, the product formation step, determines the rate of reaction according to
Michaelis-Menten assumption, the rate can be expressed as:
rP k 9 [ ES ]
(1)
[ E0 ] [ E ] [ ES ] [ EI ] [ ESI ]
(2)
k 9 [ ES ]
rP
[ E0 ] [ E ] [ ES ] [ EI ] [ ESI ]
(3)
Ks
K 2 [ E ][S ]
[ E ][S ]
[ ES ]
K1
[ ES ]
KS
(4)
KI
K 4 [ E ][ I ]
[ E ][ I ]
[ EI ]
K3
[ EI ]
KI
(5)
KI
k8 [ ES ][ I ]
[ ES ][ I ]
[ ESI ]
k7
[ ESI ]
KI
(6)
[ E ][ S ]
KS
rP
[ E ][ S ] [ E ][ I ] [ ES ][ I ]
[ E0 ]
[E]
KS
KI
KI
k9
[ E ][ S ]
KS
rP
[ E ][ S ] [ E ][ I ] [ E ][ S ][ I ]
[ E0 ]
[E]
KS
KI
KS KI
k9
Eliminate [E],
[S ]
KS
rP
[ S ] [ I ] [ S ][ I ]
[ E0 ]k 9
1
KS KI KS KI
Substitute rPmax [ E0 ]k 9
rP
rPmax
[S ]
KS
[ S ] [ I ] [ S ][ I ]
1
KS KI KS KI
rP
rPmax
[S ]
K [ I ] [ S ][ I ]
K S [S ] S
KI
KI
rP
rPmax
[S ]
K [I ]
[ S ][ I ]
KS S
[S ]
KI
KI
rP
rPmax
[S ]
K S (1
[I ]
[I ]
) [ S ](1
)
KI
KI