In this experiment we determined the rate of enzyme activity using free and immobilized enzyme. And our aim in this experiment, notice the difference between them.
Distilled water, Buffer solution, Tube, Pipette, Oil, Alcohol, Water bath, Beaker, Enzyme (Candida rugosa lipase)
Immobilized enzyme: 0.012 g Immobilized enzyme (Candida rugosa lipase) was taken and 1 ml. olive oil was added. Then 3 ml. 25 mM KP buffer was added. This solution was placed water bath at 37 C for 30 min. After immobilized enzyme had been centrifuged, supernatant was taken. And 5 ml. alcohol (ethanol) and a few drops phenolphthalein were added. The time was recorded which is taken from the centrifuged to alcohol added. After that it was titrated with 0.05 M KOH. Finally titrated volume was recorded. Free enzyme: 0.2 ml. free enzyme was weighted then 1 ml .olive oil and 3 ml. 25 mM KP buffer were added. It was placed water bath at 37 C for 30 min. After that 5 ml ethanol and a few drops phenolphthalein were added and titrated with 0.05 M. KOH. Finally titrated volume was recorded.