Lecture 1 - enzyme kinetics

Enzyme kinetics case study genetic engineering of

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Unformatted text preview: onstants for selected Codon 222 mutants Codon 222 Kcat (S-1) Km (M) Kcat / Km (M-1 S-1) Met (wild-type) Cys Ser Leu 50 84 27 5 1.4 x 10-4 4.8 x 10-4 6.3 x 10-4 2.6 x 10-4 36x104 20x104 4x104 2x104 Enzyme Kinetics! Two measures of enzyme power ! 1. Kcat = reflection of the organic chemistry of the active site measure of efficieny Kcat = Vmax E Kcat/Km = Enzyme rate constant (higher the rate constant - the better the enzyme performs, greater power) Higher the Kcat, the lower the turnover time, and the more powerful the E. concentration at half the Vmax value measure of proficiency 2. Km = measure of how [S] affects E-function Km = independent of enzyme concentration, analogous to infinity ..describes the binding of the substrate to the enzyme (based on the chemical properties of the active sites - not enzyme concentration) Enzymes with lower Km value more powerful. Enzyme Kinetics! Case Study - Genetic Engineering of Subtilisin! Table II: Kinetic Constants for selected Codon 222 mutants Codon 222 Kcat (S-1) Km (M) Kcat / Km (M-1 S-1) Met (wild-type) Cys Ser Leu 50 84 27 5 1.4 x 10-4 4.8 x 10-4 6.3 x 10-4 2.6 x 10-4 36x104 20x104 4x104 2x104...
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