10-7-0+Chem+of+Enzyme.ppt

Victo Henri proposed that an enzyme combines with its substrate molecule to form the ES complex as a necessary step in enzyme catalysis 1913 Leonor Michaelis and Maud Menten postulated that the enzyme first combines reversibly with its substrate to form ES complex in a relatively fast reversible step First step: E+S ES V∝[ES] Fast step E + S E S E + P k 1 k -1 k 2 k -2 E S E + P k 2 k -2 Second step: Slower than (1) (1) (2) Pre-steady state(前稳态): when the enzyme is first mixed with a large excess of sustrate, there is an initial period called the pre-steady state during which the concentration of the ES complex build up steady state(稳态): the reaction quickly achieves a steady state in which [ES] (and the concentration of any other intermediates) remains approximately constant overtime steady state kinetics (稳态动力学): analysis of the relationship between steady-state rate and [S] is referred to as steady-state kinetics. Michaelis and Menten equation concerned with this kinetics. 二、the relationship between [S] and Enzymatic Reaction Rate can be expressed quantitatively E + S E S E + P k 1 k -1 k 2 k -2 k1 K-1 k2 At early time in the reaction, Vo is determined by the breakdown of ES to give product, which is determined by [ES] Vo = k2[ES] P→0 (negligible) ∵ [S][Et]; [S][ES]; [Et]: total enzyme concentration [Ef]: free enzyme concentration Then [Ef] = [Et]-[ES] Rate of ES formation = d[ES] dt = k1{[Et] – [ES]}[S] Rate of ES breakdown = -d[ES] dt = k-1[ES] + k2[ES] The initial rate of reaction reflects a steady state in which [ES] is constant, i.e the rate of formation of ES is equal to its rate of breakdown. This is called the steady-state assumption Then, k1{[Et] – [ES]}[S] = k-1[ES] + k2[ES] {[Et]-[ES]}[ES] k-1 + k2 [ES] k1 = K-1 + k2 k1 is defined as the Michaelis-Menten constant, Km {[Et]-[ES]}[ES] [ES] Km