Page 1 Chapter 5 (part 3) Enzyme Kinetics (cont.) Page 2 Chapter 5 (part 3) Enzyme Kinetics (cont.) Michaelis-Menton E + S ES E + P â€¢V max â€¢K m â€¢K cat â€¢K cat /K m k 1 k -1 k cat â€¢Vo = Vmax [S] Km + [S] â€¢V max â€¢K m â€¢k cat â€¢k cat /K m Page 3 Chapter 5 (part 3) Enzyme Kinetics (cont.) Michaelis-Menton E + S ES E + P â€¢V max â€¢K m â€¢K cat â€¢K cat /K m k 1 k -1 k cat â€¢Vo = Vmax [S] Km + [S] â€¢V max â€¢K m â€¢k cat â€¢k cat /K m How do you get values for V max , K m and k cat ? â€¢ Can determine K m and V max experimentally â€¢ Km can be determined without an absolutely pure enzyme â€¢ K cat values can be determined if V max is known and the absolute concentration of enzyme is known (V max = k cat [E total ] Page 4 Chapter 5 (part 3) Enzyme Kinetics (cont.) Michaelis-Menton E + S ES E + P â€¢V max â€¢K m â€¢K cat â€¢K cat /K m k 1 k -1 k cat â€¢Vo = Vmax [S] Km + [S] â€¢V max â€¢K m â€¢k cat â€¢k cat /K m How do you get values for V max , K m and k cat ? â€¢ Can determine K m and V max experimentally â€¢ Km can be determined without an absolutely pure enzyme â€¢ K cat values can be determined if V max is known and the absolute concentration of enzyme is known (V max = k cat [E total ] Lineweaver-Burke Plots (double reciprocal plots) â€¢Plot 1/[S] vs 1/V o â€¢L-B equation for straight line â€¢X-intercept = -1/Km â€¢Y-intercept = 1/Vmax â€¢Easier to extrapolate values w/ straight line vs hyperbolic curve Page 5 Chapter 5 (part 3) Enzyme Kinetics (cont.) Michaelis-Menton E + S ES E + P â€¢V max â€¢K m â€¢K cat â€¢K cat /K m k 1 k -1 k cat â€¢Vo = Vmax [S] Km + [S] â€¢V max â€¢K m â€¢k cat â€¢k cat /K m How do you get values for V max , K m and k cat ? â€¢ Can determine K m and V max experimentally â€¢ Km can be determined without an absolutely pure enzyme â€¢ K cat values can be determined if V max is known and the absolute concentration of enzyme is known (V max = k cat [E total ] Lineweaver-Burke Plots (double reciprocal plots) â€¢Plot 1/[S] vs 1/V o â€¢L-B equation for straight line â€¢X-intercept = -1/Km â€¢Y-intercept = 1/Vmax â€¢Easier to extrapolate values w/ straight line vs hyperbolic curve B B B B B B B 0 0.05 0.1 0.15 0.2 0.25 012345 678910 Vo [S] [S] Vo 0.5 0.075 0.75 0.09 20.152 40.196 60.21 80.214 10 0.23 V max Km Km ~ 1.3 mM Vmax ~ 0.25Read More

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