Experiments of Immobilized Enzymes Using Liver Homogenate

Abstract

In the present study, I immobilized the liver homogenate from a pig on the alginate gel beads. A large quantity of the homogenate-immobilized beads were prepared easily with alginate and calcium chloride solutions.

The beads were milky white and the suspension was clear and not colored by the blood pigments for at least 4 days. So, the beads and their suspension were suitable for detection of color developed by enzyme reaction. When the beads were kept in dark at 5°C for 3 days, the catalase activity on the beads suspended in 0.15 mol/l sodium chloride solution retained 84% of the original activity, and that in 0.1 mol/l Tris buffer solution (pH7.4) retained 75%. Thus, the beads were no problem for experiments when they were used within 1-2 days after preparation.

I measured successfully the activity of catalase, alcohol dehydrogenase and alginase using the beads; the catalase activity was measured in various temperatures by trapping 02 evolved into a special test tube which was newly devised in the present study. Alcohol dehydrogenase activity was detected by the coluring of beads developed with INT reagent. Urea formed by alginase was decomposed into ammonia by added urease, then the latter was detected with Nessler’s reagent on filter paper.

It was found that the enzyme-immobilized beads were more suitable than homogenate or small piece of liver for some experiments in biology in high school.