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1. The document describes using the equilibrium approach and quasi-steady state approach to derive a rate expression for a reaction sequence involving an enzyme (E), substrate (S), and products (P1 and P2). 2. It then provides kinetic constants for the enzyme fumarase and uses the quasi-steady state approach to calculate (a) the Michaelis constant Km and (b) the initial rate of product formation for a given enzyme and substrate concentration. 3. The key steps are deriving rate expressions v = k5(ES)2 for the reaction sequence using both approaches, and then applying the expressions and kinetic constants for fumarase to calculate Km = 45 x 10-6 M

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alfi hendry
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1K views

CXX

1. The document describes using the equilibrium approach and quasi-steady state approach to derive a rate expression for a reaction sequence involving an enzyme (E), substrate (S), and products (P1 and P2). 2. It then provides kinetic constants for the enzyme fumarase and uses the quasi-steady state approach to calculate (a) the Michaelis constant Km and (b) the initial rate of product formation for a given enzyme and substrate concentration. 3. The key steps are deriving rate expressions v = k5(ES)2 for the reaction sequence using both approaches, and then applying the expressions and kinetic constants for fumarase to calculate Km = 45 x 10-6 M

Uploaded by

alfi hendry
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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1.

Consider the following reaction sequence:

Develop a suitable rate expression for production formation [ v = k5 (ES)2 ] by using:


(a). the equilibrium approach, and
(b). the quasi-steady-state approach

Answer

(a). equilibrium approach


k1 [S][E] = k2 [ES]1 ..…(1)
k3 [ES]1 = k4 [ES]2 .…(2)

[E0] = [E] + [ES]1 + [ES]2 ..…(3)

From (2)

𝐾4
[ES]1 = (𝐸𝑆)2 .…(4)
𝐾3

𝐾2 𝐾4 [𝐸𝑆]2
[E] = .…(5)
𝐾1 𝐾3 [𝑆]
From (1) + (4)
Substitute (4) and (5) into (3)

𝑘2 𝑘4 +𝑘1 𝑘4 (𝑆)+𝑘1 𝑘3 (𝑆)


(E)0 = [ ](ES)2
𝑘1 𝑘3 (𝑆)
𝑘1 𝑘3 (𝐸0 )(𝑆)
(Es)2 =
𝑘2 𝑘4 +𝑘1 𝑘4 (𝑆)+𝑘1 𝑘3 (𝑆)
𝑘5 (𝐸0 )(𝑆) 𝑟2 𝑟4
Rate = dimana km1 = , Km2 = ,
𝑘𝑚2 +(𝑘𝑚1 +𝑆)+𝑘1 𝑘3 (𝑆) 𝑟1 𝑟3

b. Quasi-stedy-state approach
𝑑 (𝐸𝑆)1
= 𝑘1 (𝐸)(𝑆) − 𝑘2 (E𝑆)1 + 𝑘4 (𝐸𝑆)2 − 𝑘3 (E𝑆) ...(6)
𝑑𝑡
𝑑 (𝐸𝑆)2
= 𝑘3 (𝐸𝑆) − 𝑘4 (E𝑆)2 − 𝑘5 (𝐸𝑆)2 …(7)
𝑑𝑡

From (7)
𝑘4 + 𝑘3
(Es)1 = (ES)2 …(8)
𝑘3

From (6) + (8)

𝑘2 𝑘4 +𝑘2 𝑘5 +𝑘3 𝑘5
(E)0 = [ ](ES)2 …(9)
𝑘1 𝑘3 (𝑆)
𝑘2 𝑘4 +𝑘2 𝑘5 +𝑘3 𝑘5 +𝑘1 𝑘4 (𝑆)+𝑘1 𝑘3 (𝑆)+𝑘1 𝑘3 (𝑆)
(E)0 = [ ](ES)2
𝑘1 𝑘3 (𝑆)
𝑘1 𝑘3 (𝐸0 )(𝑆)
(ES)2 = [ ]
𝑘2 𝑘4 +𝑘2 𝑘5 +𝑘3 𝑘5 +𝑘1 𝑘4 (𝑆)+𝑘1 𝑘3 (𝑆)+𝑘1 𝑘3 (𝑆)
𝑘1 𝐸0 𝑆
Rate = 𝑘 𝑘
(𝑘𝑚2 5⁄𝑘3)(𝑘𝑚1 +𝑆) + 5 ⁄𝑘 +𝑆
1
𝑘 𝑘
dimana Km1 = 𝑘2 and Km2 = 𝑘4
1 3
2. The enzyme, fumarase, has the following kinetic constants:

a. What is the value of the Michaelis constant for this enzyme?


b. At an enzyme concentration of 10-6 M, what will be the initial rate of product formation at
a substrate concentration of 10-3 M?

Answer
Asumsi quasi stady state

𝑑 (𝐸𝑆)
= k1 [S][E] – k-1 [ES] - k2 [ES] = 0 …..(1)
𝑑𝑡
k[S][E] = k-1 [ES] + k2 [ES]
k[S][E] = [ES] (k1 + k2)
𝑘1 [𝑆][𝐸]
[ES] = …..(2)
𝑘2 +𝑘−1

NERACA MASSA ENERGI TOTAL

[E] = [E0] – [ES] …..(3)


Subtitusi persamaan 1 dan 2
𝑘1 [𝑆]([𝐸0 ]−[𝐸𝑆]
[ES] =
𝑘2 +𝑘−1

𝑘1 [𝑆][𝐸0 ]−𝑘1 [𝑆][𝐸𝑆]


[ES] =
𝑘2 +𝑘−1
[ES] (k2 +k-1) = 𝑘1 [𝑆][𝐸0 ] − 𝑘1 [𝑆][𝐸𝑆]

[ES] (k2 +k-1) + 𝑘1 [𝑆][𝐸𝑆] − 𝑘1 [𝑆][𝐸0 ]

[ES] (k2 +k-1 + 𝑘1 [𝑆]) = 𝑘1 [𝑆][𝐸0 ]

[ES] (k2 +k-1 + 𝑘1 [𝑆]) = 𝑘1 [𝑆][𝐸0 ]

: 𝑘1
(𝑘2 +k−1 + 𝑘1 [𝑆])
[ES] = [S][E0]
𝑘1
[𝑆][𝐸0 ]
[ES] = (𝑘2 + k−1 + 𝑘1 [𝑆])
𝑘1

[𝑆][𝐸0 ]
[ES] = 𝑘 …..(4)
2 k−1 +[𝑆]
+
𝑘1

Laju Pengukuran Produksi


𝑑[𝑝]
V = rp = = k2 [ES] …..(5)
𝑑𝑡
Subtitusi persamaan 4 dan 5
𝑑[𝑝] [𝑆][𝐸0 ]
V= = k2 = 𝑘2 + k−1
𝑑𝑡 +[𝑆]
𝑘1

Berdasarkan rumus yang ada


𝑑[𝑝] 𝑉𝑚 . [𝑆]
V= = k2 =
𝑑𝑡 km + [𝑆]

Diperoleh
𝑘2 + k−1
Km =
𝑘1
Dan
Vm = k2 [E0]
A. Nilai Dari Konstanta Michaelis (Km)

𝑘2 + k−1
Km =
𝑘1
2 4
10 +(4,4 ×10 )
= 9
10
(44+1)×103
=
109
= 45 x 10-6

B. Pada [E0] =10-6 M


[S] = 10-3 M
Vm =k3 [E0]
Vm = 10 -3
𝑉𝑚 [𝑆]
Vp =
𝑘𝑚+[𝑆]

10−3 [10−3 ]
=
(45 ×10−6 )+10−3

10−6
=
(45 ×10−6 )+1000 ×10−6

10−6
=
(45 ×1000) ×10−6

1
=
1045

= 0,00095 atau 9,4 x 10-4

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