Purpose:

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.

Materials:

Distilled water, Buffer solution, Tube, Pipette, Oil, Alcohol, Water bath, Beaker, Enzyme (Candida rugosa lipase)

Procedure:

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.

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