A specified amount of lipase Candida Rugosa (optimally 1 g)
was measured into a flask and 10 ml of water was added.
The mixture was stirred at 200 rpm using a magnetic stirrer
for about 15 min. To this mixture, 0.5 ml of a 0.5 M NaF solu-
tion and the activated carbon was added. The flask was re-
moved from the stirrer and left sealed at room temperature
for 24 h. The flask was then uncapped and was incubated in
a water bath at 37 C for about 48 h. The powder was washed
with 100 ml of distilled water in a 250 ml flask for 1 h at a mix-
ing speed of 500 rpm. The mixture was then filtered. The acti-
vated carbon with the lipases was dried again at 37 C for 48 h.
The granular carbon was stored at 4 C until use. Based on the
degree of immobilization tests, about 95% of the enzyme was
immobilized in this procedure. The actual enzyme loading was
determined at 500 mg of lipase Candida Rugosa 10 g of acti-
vated carbon.