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    CXX

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    alfi hendry
    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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    CXX

    Uploaded by

    alfi hendry
    1K views5 pages
    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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    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

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    1K views5 pages

    CXX

    Uploaded by

    alfi hendry
    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

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
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    The text discusses how to derive rate expressions for an enzyme-catalyzed reaction using the equilibrium approach and quasi-steady state approach. It also shows the steps to derive the Michaelis-Menten equation.

    Using the equilibrium approach, the rate expression is derived by setting up equilibrium expressions and substituting them into the total enzyme concentration equation. Using the quasi-steady state approach, differential rate equations are set up and solved to derive the rate expression.

    The steps shown are: 1) Write the differential rate equation for ES formation. 2) Assume quasi-steady state for ES. 3) Substitute the quasi-steady state expression into the total enzyme and substrate equations. 4) Isolate [ES] and substitute into the rate equation to obtain the Michaelis-Menten equation.

    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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