Description: A peroxidase enzyme. which was extracted from a Brassica compestris ( Brassica rapa ) . is tested under assorted conditions in temperature. pH degree. and competitory inhibitor ( hydroxylamine ) .
In order to find the belongingss of an enzyme. a peroxidase enzyme was extracted from a Brassica compestris ( Brassica rapa ) and tested under assorted temperatures. pH degrees. and by a competitory inhibitor ( hydroxylamine ) . The enzyme activity was measured in assorted ways depending on the activity. Temperature effects showed the sum of activity and determined the optimum pH degree. The spectrometer showed the optical density units at 500nm and determined the optimum temperature. The temperatures tested where 4?C ( about refrigerator temperature ) . 23?C ( about room temperature ) . 32?C. and 48?C. The pH degrees tested were ; pH3. pH5. pH7. and pH9. The competitory inhibitor used was Hydroxylamine. These trials resulted in the ability to find the optimum temperature: 32?C because it closest to organic structure temperature. The best pH degree was found to be pH5.
An enzyme is a protein that serves as a accelerator. which is a chemical agent that changes the rate of a reaction without being consumed by the reaction. The enzyme dramatically speeds up reactions by take downing the activation energy barrier. Every enzyme has a alone form because the substrates that bond to it are all different forms. It fits like a lock and cardinal. Once the substrate enters the active site. the enzyme somewhat changes form in order to lock in the substrate and holds it in topographic point. Enzymes allow beings to populate by increasing the rate of a chemical reaction. The peroxidase enzyme catalyzes the reaction of the formation of O. Assorted temperatures and pH degrees will be tested in order to find the optimum environment for the peroxidase enzyme.
Temperature: If the optimum human organic structure temperature is 36?C. so the enzyme’s optimum temperature should be similar.
Boiling: If most proteins are denatured when they are heated to a temperature above 70?C. so the enzyme will be dead when boiled.
pH: Because the beings are highly sensitive to alterations in pH degrees. the peroxidase enzyme excessively will be highly sensitive.
Hydroxylamine: Because hydroxylamine has a similar form to the substrate. H peroxide. it will bond to portion of the enzyme and barricade the substrate from come ining. This would be measured by lessening of optical density units.
MATERIALS AND METHODS
Please see commendations for inside informations: ( Dolphin. 2005 )
*Changes to lab protocol:
page 81. Fixing an infusion incorporating peroxidase ; measure 1. 1 gram alternatively of 10.
page 81. Alternatively of adding 100ml at one time. add 30ml. swot and so add 70ml.
page 81. Standardizing the Amount of Enzyme: label 3 beakers alternatively of 4.
page 87. Alternatively of no control. there was a control of tubing that had a pH5. 2. 0 H2O2mL. 0mL infusion. 0mL hydroxylamine treated infusion. 1. 0mL Guaiacol. and a entire volume of 8mL.
Please refer to included tabular arraies and graphs.
The followers are attached:
Table 1/Figure 1: Volume of Turnip Extract
Table 2/Figure 2: Temperature Graph
Table 3/Figure 3: Temperature Derivative Graph
Table 4/Figure 4: pH Graph
Table 5/Figure 5: pH Derivative Graph
Table 6/Figure 6: Boiled Infusion
Table 7/Figure 7: Hydroxylamine Graph
Based on the graph ( Figure 2 ) . it was determined that the most optimum temperature for the peroxidase enzyme was 32?C. It was besides determined that the enzyme maps to its highest potency at the pH 5 degree. That is shown in Figure 4. In add-on. Figure 6 proved that the enzyme becomes denaturized when heated above 70C. Last. the enzyme inhibitor. hydroxylamine. resulted in a lessening of optical density units.
The hypothesis that was originally stated sing the assorted temperatures needs to be somewhat modified. The optimum temperature is non precisely the same to the human organic structure nevertheless it is comparatively close ( four grades difference ) .
The hypothesis for the consequence of assorted United States Public Health Service on the activity of the peroxidase enzyme was non clear plenty and did non foretell any consequences. Based on the experiment. the best pH for the peroxidase enzyme is pH 5.
Meanwhile. the hypothesis for the consequence of boiling the infusion was right. The consequences obtained in the experiment supported the original hypothesis that when proteins ( which an enzyme is ) are heated to a temperature above 70?C. the enzyme will be dead when it is boiled.
Based on the effects of the hydroxylamine treated infusion. the original hypothesis is once more accepted. Because the inhibitor. hydroxylamine. blocked the substrate of H peroxide from come ining the active site. the optical density units decreased compared to the normal extract optical density units.
Further work that would be needed to prove accounts would include a more assortment of temperatures every bit good as other enzymes and compare the consequences between assorted enzymes.
Dolphin. Warren D. 2005. Biological Investigations. Lab Topic 7 Determining the Properties of an Enzyme. erectile dysfunction. Kent A. Peterson. pp. 79-89.
New York. New york: McGraw-Hill