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Effect Of Temperature On Enzyme Activity
Role of the experiment
The enzyme is extracted from turnips by homogenization in buffer. The broken cells release Peroxidase and other enzymes in the buffer. The enzyme is separated from the cellular debris by filtration or centrifugation. The presence of peroxidase is then tested by use of H202 and a compound known as Guaiacol, which if peroxidase is present, it will breakdown peroxide and the oxygen released will react and produce a brown compound. In this experiment, five test tubes are used. The first tube, labeled 0, will be the control. All the ingredients that are required for the reaction except the extract will be placed in tube 0. The successive test tubes will have the extract (Peroxidase). The test tubes will then be placed in water baths heated to different temperatures (20-50o) and thereafter observed. The rate of brown pigment formation will indicate whether the rate is affected by varying temperature.
The reaction for the conversion of hydrogen peroxide to water and Oxygen occurs spontaneously, but slowly in the absence of Peroxidase. Thus in the control, the rate of brown pigment formation is very slow. However, in the heated water baths, the rate of color formation increases with increase in temperature, but in the 500test tube it will stop after some minutes.
The rate of enzyme-catalyzed reactions increases with increase in temperature range. A 10 degree rise in temperature will increase the activity of enzymes by 50-100%. However, most enzymes are adversely affected by high temperatures. Most animal enzymes lose their structure and become denatured at temperatures above 40o. For any enzymatic experiment to be a success thus, temperatures below 40oare to be maintained.