BME 20100 Lecture Notes - Lecture 6: Biocatalysis, Enzyme Kinetics, Isozyme
27 Nov 2017
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Kinetics: study of the rate at which compounds react. (2) record rate of substrate disappearance/product formation. (3) plot initial velocity vs. substrate concentration. Both absorb at 260nm that release fluorescent product. Fluorimetric assays - uses naturally fluorescent molecules or synthetic substrates. Luminescence reactions - releases light upon enzymatic activity. Immunochemical assays - monoclonal antibodies to specific enzymes. K(cat) - turnover number, how many substrate molecules one enzyme molecule. K(m) - michaelis constant, approximate measure of a substrate affinity for an. During steady state, vmax occurs when enzyme is in es complex and is can convert per sec enzyme dependent on breakdown of that complex. Specificity constant-rate constant for the conversion es to ep. Rate can"t be faster than diffusion-controlled encounter of e and s. Kinetic mechanism: order of binding of s and release of p. When 2 or more reactants are involved, enzyme kinetics allows to distinguish between different mechanisms. All s must bind to e before p is released.
