1. During the test of an enzyme catalyzed reaction following data were recorded: T(C) 30 30 30 0.033 0.02 30 0.01 30 49.6 49.6 49.6 49.6 49.6 0.005 0.1 0.033 0.01 0.0067 S 0.1 0.005 (mmol/ml) V 2.63 1.92 1.47 0.96 0.56 5.13 3.7 1.89 1.43 1.11 (mmol/ml- min) Eo (g/l) 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.6 (i) Determine the Michalis-Menten constant for the reaction at 30C and 49.6C. (ii) When same enzyme is immobilized, does immobilized enzyme show diffusion limitations? T(C) Vmax (mmol/min 106) Soluble Immobilized 0.8 25 0.62 35 1.42 1.4 45 3.6 3 55 8 6.2 65 16 11 (iii) also determine the activation energy (AE cal/ g.mole) for the soluble and immobilized enzyme reaction? (5+5+15=25)