Essay --

The plebeian extract was retrieved by homogenizing the bovine vim tissue. The total enzyme in the muscle tissue domiciliate be illustrated by the total application present in the crude extract, which was 5500 200 units. beside the 40% ammonium ion sulfate step was per anatomyed. Ammonium sulfate was tardily added to a beaker that contained the crude extract until a 40% saturation take was reached. After centrifugation the pellet (which contained low soluble proteins and other cellular debris) was cast aside and the supernatant was kept. The total protein was 124 6 mg and the specific activity was 4500 three hundred units. This resulted in a specific activity of 36 4 units/mg. The knuckle under was metrical to be 82 9% and the purification factor was 1.4 0.3. there was not much of an increase in the excellence, that there was a decrease in the total protein and the specific activity as compared to the crude extract which suggests that that unwanted protein was re moved during this purification step. Performing the 60% ammonium sulfate step further purified the 40% supernatant. The ammonium sulfate and was added to the 40% supernatant until a 60% saturation level was reached. This was then centrifuged, but this time the pellet was kept while the supernatant (which contained material such as nucleic acids and sugars) was discarded. The total protein recovered was 56.4 0.4 mg and the total activity was 4140 50 units. This produced a specific activity of 73 1 units/mg. The yield was calculated to be 76 4% and the purification factor was 2.8 0.3. Each of these locomote was consistent with the trends present in the theoretical values data because the purity increased. This indicates that the unwanted material was removed so that the 60% pellet can be us... ...olecular weight). By using the protein concentration and enzyme concentration of E10 and E11 the specific activity values can be calculated. The specific activity of E10 is 497.68 u nits/mg and E11 is 516.6 units/mg. The m form is assumed to have a lower binding proportion (lower Km) so this would elute out first thus we can reason that the m form is present in the E10 fraction and the h form is present in the E11 fraction. The isozymes are assumed to have the same molecular weight but varying charge strength. With this information an ion reciprocation towboat can be performed in order to help separate the isozymes. For an anion exchange column the beads would be positive. By making one of the isozymes much negative it would bind to the beads while the other isozyme would elute out. A gel column can then be performed to confirm the comportment of different isozymes.