Discussion
Fold purification was calculated from specific activity. From sample A to B there was the addition of MgCl2 which denatured some proteins that were removed in the pellet. The second step of purification was even more successful with a 4.1 fold increase in purification, it was achieved by denaturing the non-recombinant E.Coli proteins. The Second step was much more beneficial to the total purification but the combination of the two steps achieved an even better purification of 4.6 fold. From this assay we can see that the correct enzyme has been successfully purified. Considering the SDS-PAGE results we can confidently say that the purification steps were effective at removing a large proportion of E.Coli proteins. However, not all E.Coli proteins have been removed as shown by the SDS-PAGE gel photo and further purification steps would be needed to have a pure Arginase protein. The increase in specific activity was also shown in the SDS-PAGE gel as the band in sample C was much thicker than in sample A and B showing that there was much more Arginase present in sample C. …show more content…
When they are bound, they can't have any effects on the body and they are removed from the body (Edta, 2016). So this means that adding EDTA has removed all the metal ions that were free in solution. As the specific activity of the Arginase is zero when EDTA is added it shows that Arginase required metal ions in solution to be active. This means that Arginase is a metallo-enzyme as it requires a metal ion to be
Fold purification was calculated from specific activity. From sample A to B there was the addition of MgCl2 which denatured some proteins that were removed in the pellet. The second step of purification was even more successful with a 4.1 fold increase in purification, it was achieved by denaturing the non-recombinant E.Coli proteins. The Second step was much more beneficial to the total purification but the combination of the two steps achieved an even better purification of 4.6 fold. From this assay we can see that the correct enzyme has been successfully purified. Considering the SDS-PAGE results we can confidently say that the purification steps were effective at removing a large proportion of E.Coli proteins. However, not all E.Coli proteins have been removed as shown by the SDS-PAGE gel photo and further purification steps would be needed to have a pure Arginase protein. The increase in specific activity was also shown in the SDS-PAGE gel as the band in sample C was much thicker than in sample A and B showing that there was much more Arginase present in sample C. …show more content…
When they are bound, they can't have any effects on the body and they are removed from the body (Edta, 2016). So this means that adding EDTA has removed all the metal ions that were free in solution. As the specific activity of the Arginase is zero when EDTA is added it shows that Arginase required metal ions in solution to be active. This means that Arginase is a metallo-enzyme as it requires a metal ion to be