James Lee Enzyme Kinetics Solution

BK10110302 V.
PRASARNTH RAAJ VEERA RAO BIOPROCESS

Solution 2.1
Total volume = 44 + 5 + 1= 50ml
From the graph the equation obtained
y=0.033 + 0.03a 1
m= 0.033 mol / ml . min
a) Activity of the - glucosidase

0.033 x 50= 1.65 mumol / min
i)
= 1.65mumol/min
0.1 mg/ml x 0.1ml
= 165 units/mg protein

ii)
= 1.65 mumol/min
1ml of enzyme
= 1.65 units/ml of enzyme
b) Initial rate of reaction 0.033 mumol/ mL.min
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
BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Solution 2.2
a) Michaelis-Menten approach
The rate of product formation.
d[p]
dt
Since the enzyme is preferred,
Make E as the subject,
Since forward reaction = backward reaction.
( )
ubstitute
into
:
[(
(
Make
) (
)( )
)] ( )
)( )
)( )
as a subject:
(
)( )
)( )

(
)( )
)( )
[(
(
(
(
ub
into
)( )
)( )
)( )
)( )( )
)( )( )
as a subject,
(
)(
( )( ))
)
(
into
)( )( )
)( )( )
)(
)
(
ub
)( )
)
Make
)( )]
( )( ))
)( )( )
( )
( )( )

d[p]
dt
Since
)( )( )
)
( )
( )( )
)( )( )
)
( )
( )( )
)
( )( )
)
( )
( )( )
b) Since [ ]
[ ]
d[p]
dt
( )( )
( )( )

Solution 2.3
(a) E+S k1
(ES)1
(ES)1 k3 (ES)2
(ES)2 k2 E+P
V=
[ ]
= k5 [ES] 2
[E0] = [E] + [ES] + [ES]2

[E] = [E0]-[ES]-[ES]2
k2 = [E] [S]
k1 [ES]1
k2 [ES]1 = [Eo] [S] [ES]1 [S]
k1
[ES]1 ( k2 + [S] ) = [E0] [S] [ES]2 [S]
k1
[ES]1 = [E0] [S] [ES]2 [S]
k2 + [S]
k1
k4 = [ES]1
k3
[ES]2
k4 [ES]2 = [E0] [S] [ES]2 [S]
k3
k2 + [S]
k1
[ES]2 ( k2 k4 + k4 [S] ) = [E0] [S] [ES]2 [S]
k1k3
k3
[ES]2 ( k2 k4 + k4 [S] + [S] )= [E0] [S]
k1 k3
k3
[ES]2 = [E0] [S]
k2 k4 + k4 [S] + [S]
k1 k3
k3

V= d [P] = k5 [E0] [S]
dt
k2 k4 + k4 [S] + [S]
k1 k3
k3
=
Vm [S]
k2 k4 + k4 [S] + [S]
k1 k3
k3

Solution 2.4
a) Michaelis-Menten approach
The rate of product formation.
d[p]
dt
Make E as the subject,
Since forward reaction = backward reaction.
( )
ubstitute
into
:
[(
Make
) (
)( )
)] ( )
)( )
)( )
as a subject:
(
)( )
)( )

(
)( )
)( )
[(
(
(
(
ub
into
)( )
)( )
)( )
)( )( )
)( )( )
as a subject,
(
)(
( )( ))
)
(
into
)( )( )
)( )( )
)(
)
(
ub
)( )
)
Make
)( )]
( )( ))
)( )( )
( )
( )( )

d[p]
dt
Since
)( )( )
)
( )
( )( )
)( )( )
)
( )
( )( )
)
( )( )
)
( )
( )( )
b) Briggs-Haldane approach
The rate of product formation,

d(p)
dt
Make as a subject,

Substrate consumption,
d(
( )( )
dt
d(
dt
ubstitute
into
)(
)(
)(
)( )
( )
( )
( )
( )
ubstitute
)( )
( )( )
)( )
( )(
( ))
( )
( )
( )(
( )
( )(
( )(
(
( ))
into
)( )
:
(
)(
)(
)(
( )
)( )
( )(
(
( ) ( )
( )
( ) ( )(
(
( )
( ))
)
( ))
( )
)( )
)(
)(
( )
( ) ( )
( ) ( )
(
ubstitute
)(
( ))
( ) ( )(
( )
)(
( )
( )
( ))
( ) ( )
( ) ( )
(
( ) ( )
( )
)(
( )
( ))
into
d(p)
dt
( ) ( )
(
( ) ( )
( )
)(
( )
( ))
v
d(p)
dt
v
(
( ) ( )
( ) ( )
( )
)(
( )
( ))

Solution 2.5
Lineweaver- Burk Plot
x-intercept= - 1
km
y-intercept= 1/ V more
Equation obtained y= 0.0172 x + 3.6342
y-intercept = 3.6342= 1/ V max
V max = 0.275
x-intercept , y= 0
0.0172x + 3.6342=0
0.0172x = -3.6342
x= -211.291
x= -1/km
km = 1/211.291
= 0.00473
Longmuir Plot
Equation obtained y= 3.3133x + 0.0191
1/Vm = m = 3.3133
Vm=0.302
y-intercept= km/Vm = 0.0191
Km = 0,0191x 0.302
= 0.00577
Eadie-Hofstee Plot
Equation obtained y= -0.0043x + 0.2645
-Km = m = -3.3133
Km=0.302

y-intercept= Vm = 0.2645
Non-Linear Regression Procedure
From the graph, Vm=0.2
Vmax = 0.1,
Km=0.0032
Data for Graph plot :

Langmuir Plot
s
s/v
0.0032 0.028829
0.0049 0.033108
0.0062 0.043357
0.008 0.048193
0.0095
0.0475
Lineweaver-Burk Plot
1/s
1/v
312.5
9.009009
204.0816 6.756757
161.2903 6.993007
125
6.024096
105.2632
5
Eadie-Hofstee Plot
v/s
v
34.6875
0.111
30.20408 0.148
23.06452 0.143
20.75
0.166
21.05263
0.2
Non-Linear Regression Plot
S
v
0.0032
0.111
0.0049
0.148
0.0062
0.143
0.008
0.166
0.0095
0.2
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
Lineweaver Burk Plot

10
9
8
7
6
5
4
3
2
1
0
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
Non-Linear Regression Procedure

0.25
0.2
0.15
0.1
0.05
0
0
0.002
0.004
0.006
0.008
0.01

Solution 2.6
Rate of product formation

( )
Enzyme is preserved,
d( )
dt
(
Assumptions:
[ ]small,
neg igib e
( )( )
)(
)(
(
(
into
))( )
)
( )
(
(
)
)
( ) )
( )
( )
(
( )
d(p)
dt
(
(
( )
into
( )
(
( )( )
(
)
Substitute
v
( )( )
Substitute equation
(
()
( )
( )
)
)

Dividing
[(
with the value of
( )
( )
]/
v s
( )

Solution 2.7
a) FCs0 - FCs + rSv = V
For Batch reactor F=0
rSv = V
[ ]
[ ]
[
]
[
= 60mol/m3.min
b) Equation obtained y = 6.3852x + 59.571

m = Vmax
= 6.3852
y- intercept = - Km = 59.571
Km = - 59.571
c) FCs0 - FCs + rSv = 0

FCs0 - FCs = - rSv = rpv
FCs0 - FCs =
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

Data :
Cs t t/ln(Cso/Cs) (Cso-Cs)/ln(Cso/Cs)
1 1
0.175322
52.42135
5 5
1.221197
72.0506
10 10
2.940141
85.26409
20 20
7.385387
103.3954
Graph :
(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

Solution 2.8
a) Km =0.01 mol/L
Cso = 3.4 x 10 -4 mol/L
Cs = 0.9 x 3.4 x 10-4
= 3.06 x 10-4 mol/L
t= 5minutes
[ ]
[ ]
( 3.4x 10-4 3.06 x 10-4) = Vmax (3.06 x 10-4)

S
0.01 + (3.06 x 10-4)
6.8 x 10-6 = Vmax ( 0.03)
Vmax = 2.27 x 10-4 mol/L-min
b) 6.8 x 10-6 x 15 = 1.02 x 10-4 mol/L

Solution 2.9
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
b) Plug - Flow @ Stead State

Km ln
+ (Cs0 - Cs )
0.03 ln
+ (10 - 0.5 ) = 780t
= rmax t
9.95899 = 780t
t = 0.0123hr
t = V/F = 0.0123
V = 0.0123 10
= 0.123liter

Solution 2.10
Km = 10g/L
rmax = 7g/L.min
a)
F=0.5L/min
Cs0=50g/L
1L
F=0.5L/min
Cs1=?g/L
1L
F=0.5L/min
Cs2=? g/L
CSTR @ Steady State

F (Cs0 - Cs ) = rpv
0.5 (50 Cs1) =
)
s
(1)
(25-0.5Cs1)(10+ Cs1)=7Cs1
250+25Cs1-5Cs1-0.5Cs12=7Cs1
0.5Cs12-13Cs1-250=0
Cs1=38.86g/L
0.5 (38.86 Cs2) =
)
s
(1)
(19.43-0.5 Cs2)(10+ Cs2)=7 Cs2

194.3+19.43Cs2-5Cs2-0.5Cs22=7Cs2
0.5Cs22-7.43Cs2-194.3 =0
Cs2=28.49g/L

b)
F=0.5L/min
Cs0=50g/L
2L
F=0.5L/min
Cs1=?g/L
CSTR @ Steady State

F (Cs0 - Cs ) = rpv
0.5 (50 Cs1) =
)
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.

Solution 2.11
a) k1 [E] [S] = k2 [ES]
[ES] = k2
k1 [E] [S]
k1 [E] [S]
k2
k3 [E] [P] = k4 [EP]

[EP] = k3
k4 [E] [P]
k5 [ES][P] = k6 [ESP]
[ESP] = k5 [ES] [P]
k6
k7 [EP] [S] = k8 [ EPS ]
[EPS] = k7 [EP] [ S ]
K8
= k7 k3 [ S ]
k8 k4 [E] [P]
From,
[ESP] = k5 [P] k2
k6k1 [E] [S]
[E0] = [E] + [ES] + [EP] + [ESP] + [EPS]
[E0] = [ES] + [ESP] + [E] + [EP] + [EPS]
[E0] = [ES] + [ESP] + [E] + [EP] +
][ ]
[ ]
[E0] = [ES] + [ESP] + [E] + [EP] + (

[ ][ ]
[E0] = [ES] + [ESP] + [E] +
[ ]
[E0] = [ES] + [ESP] + [E] [

[E0] = [ES] +
[E0] = [ES] {1 +
][ ]
][
[ ]
[ ]
+(
[ ]
(
[ ]
[
[ ]
)]
[ ]
(
[ ]
)]}
)]

[ES] =
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([ ] [ ])
(
[ ]
[ ]
)

Y = mx+c
[ ]
)
]
n ([
Y=
M=
[
X= (
] [ ]
C=
So we can plot a graph of
[ ]
)
]
n ([
vs (
] [ ]

Solution 2.14
Rate:
rp = k9CES +k10CEIS1 +k10CEIS2
---- 1
Enzyme balance:
CEo = CE + CES
---- 2
CEo = CEIS1 + CES + CE
---- 3
CEo = CEIS2 + CEI + CE
---- 4
The equilibrium reaction equations are as follows:

CE Cs / CES = k2/k1
---- 5
CECI / CEI = K4/K3
---- 7
CESCI /CEIS1 = K6/K5
---- 6
CEICS / CEIS2 = K8/K7
---- 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
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 }
CE = (K4/K3) CEI
CEI = K8/K7CS CEIS2
---- 10

From Equation 4,
CEo = CEIS2 + CEI + [(K4/K3)CI]CEI
= CEIS2 + [1 + (K4/K3)CI ]CEI
= CEIS2 + [1 + (K4/K3)CI ]( K8/K7)CS CEIS2
CEo = {1 + [1 + (K4/K3)CI ]( K8/K7)CS } CEIS2
CEIS2 = CEo / {1 + [1 + (K4/K3)CI ]( K8/K7)CS }
---- 11
From Equation 1, since rp = k9CES +k10CEIS1 +k10CEIS2,

By substituting Equation 9, 10 & 11 into Equation 1,
Therefore,
rp = k9 CEo /[( k2/k1)CS +1] + k10 CEo/ {1 + [1 + (k2/k1) Cs ]( K6/K5)CI } + k10 CEo / {1 + [1 + (K4/K3)CI
]( K8/K7)CS }

Solution 2.15
a) Based on the graphs
The y-intercept in Lineweaver Burk plot is almost the same.
Y-intercept => 3.8266; 3.6342
Whereas in Langmuir Plot
Two equations obtained
Y = 2.9883x + 0.0489
Y = 3.3133x + 0.0191
When y=0
X=
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
b) Y-intercept = 1/Vmax = 0.00489

Vmax = 1/0.00489 = 204.5 mol /L.min
Km/Vmax = 2.9883
Km=2.9883*204.5
=611mol/L
0.008
0.01

Solution 2.16
(a) E + S
ES + S
k5 E + P
(E0 ) = (E) +(ES) + (ESS)
(E) = (E0 ) (ES) (ESS) -------V=
=
(
( )
= k5 (ES) ------
)( )
)
) = (ES)(S)/ (k4 / k3)
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
(ES) ( (k2/k1) + (S)( ) + (S)2 ) =

(ES) =
(E0)(S)
(E0)(S) / (k2/k1) + (S)( ) + (S)2 --------
3
V=
=
( )
k5 (E0) (S) / (k2/k1) + (S)( ) + (S)2
Vm(S) / (k2/k1) + (S)( ) + (S)2
(b) At low substrate concentration,

1/ Vm = 3.1209
Vm = 0.3204
Km/Vm = 106.07
Km / 0.3204 = 106.7
K1m = 33.98
At high substrate concentration,
1/ Vm = 3.0574
Vm = 0.3271
1/ K1. Vm = 0.0032
1/ Km(3.0574) = 0.0032
Km = 102mol/L
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
b) Find rp for each F and s

Equation obtained y= 0.0391x + 0.1641
M= 1/Vmax = 0.0391
Vmax= 25.57 m mol/L.min
Km/ Vm = 0.1641
Km= 4.197
Solution 2.18
[SO]1 = 0.1 mol/L
[S0]2 = 0.3mol/L
[ ][ ]
[
]
[ ][
]
]
[ ][
[ ][
]
]
[
[ ][
[
]
]
[ ][ ]
[
]
V1 =
= k5[ES1]
=3.5 [ES1] --V2 =
= k6[ES2]
=2.8 [ES2] ---
[E0] = [E] + [ES1] + [ES2]

[ ][
[E0] = [E] + [ES1] +
[E0] = [ES1] + [E] (1+

[E0] = [ES1] +
[E0] = [ES1] {1 +
[
[ES1] =
[
[
]
(1+
(1+
]
(
)}
[E0] = 0.05 mol/L

Vmt = [S1]0 [S2] + K ln
[ ]
[ ]
3.5 [ES1] t = 0.1 [S1] +0.0714 ln [
3.5 [ES1] t + [S1] = 0.1 +0.0714 ln 0.1 - 0.0714 ln [S1]

3.5 [ES1] t + [S1] + 0.0644 = -0.0714 ln [S1]
ln[S1] =
[S1] =
---
[E0] = [E] + [ES1] + [ES2]

[E0] = [E] +
[ ][
[E0] = [E] (1+

[E0] =
[
]
[E0] = [E2] [
+ [ES2]
(1+
)+ [ES2]
[
(1+
Vmt = [S1]0 [S2] + KMln
)+ [ES2]
]
)+ 1]
]
2.8[ES1] t = 0.3 [S2] + 0.2207ln [
2.8[ES1] t + [S2] = 0.3 + 0.2207ln 0.3 0.2207ln [S2]

2.8[ES1] t + [S2] 0.0343 = 0.2207ln [S2]
ln[S2] =
[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

Solution 2.20
a. Write the kinetic model.
Since the Michaelis constant KM is not affected by the presence of the inhibitor (which has shown on
the given table); then this enzyme reaction is noncompetitive inhibition reaction.
Kinetic Model:
k 1, k 2
E S
ES
k 3, k 4
E I
EI
k 5, k 6
EI S
EIS
k 7 ,k 8
ES I
ESI
k9
ES
EP
b. Derive the rate equation. State the assumptions.

Assumptions:
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.
The two equilibrium reactions,
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)
The enzyme balance gives
[ E0 ] [ E ] [ ES ] [ EI ] [ ESI ]
(2)
k 9 [ ES ]
rP
[ E0 ] [ E ] [ ES ] [ EI ] [ ESI ]
(3)
Divide (1) by (2),

Applied Law of mass action,
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)
Substitute (4), (5), (6) into (3),
[ 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
Multiply numerator and denominator by Ks,
rP
rPmax
[S ]
K [ I ] [ S ][ I ]
K S [S ] S
KI
KI

Rearranging,
rP
rPmax
[S ]
K [I ]
[ S ][ I ]
KS S
[S ]
KI
KI
rP
rPmax
[S ]
K S (1
[I ]
[I ]
) [ S ](1
)
KI
KI

James Lee Enzyme Kinetics Solution

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James Lee Enzyme Kinetics Solution

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BK10110302 V.

PRASARNTH RAAJ VEERA RAO BIOPROCESS

a) Activity of the - glucosidase

= 165 units/mg protein

b) Initial rate of reaction 0.033 mumol/ mL.min

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Since the enzyme is preferred,

Make E as the subject,

Since forward reaction = backward reaction.

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

[E0] = [E] + [ES] + [ES]2

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Since the enzyme is preferred,

Make E as the subject,

Since forward reaction = backward reaction.

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

The rate of product formation,

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Data for Graph plot :

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Lineweaver Burk Plot

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Non-Linear Regression Procedure

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

Rate of product formation

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

with the value of

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

b) Equation obtained y = 6.3852x + 59.571

c) FCs0 - FCs + rSv = 0

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

( 3.4x 10-4 3.06 x 10-4) = Vmax (3.06 x 10-4)

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

b) Plug - Flow @ Stead State

+ (10 - 0.5 ) = 780t

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

CSTR @ Steady State

0.5 (38.86 Cs2) =

(19.43-0.5 Cs2)(10+ Cs2)=7 Cs2

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

CSTR @ Steady State

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

k3 [E] [P] = k4 [EP]

[E0] = [ES] + [ESP] + [E] + [EP] + (

[E0] = [ES] + [ESP] + [E] +

[E0] = [ES] + [ESP] + [E] [

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

BK10110302 V.PRASARNTH RAAJ VEERA RAO BIOPROCESS

CEo = CEIS1 + CES + CE

CEo = CEIS2 + CEI + CE

The equilibrium reaction equations are as follows:

CECI / CEI = K4/K3

CESCI /CEIS1 = K6/K5