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Shoumit Sarkar

An experiment investigating activity of enzymes at different temperatures Enzymes are mainly proteins, that catalyze (i.e., increase the rates of) chemical reactions. In enzymatic
reactions, the molecules at the beginning of the process are called substrates, and the enzyme converts them into different molecules

Aim
To test whether temperature has an effect on the reaction between amylase and hydrogen peroxide.

Hypothesis
As enzymes have an optimum temperature, the enzymes would work best and have the highest rate of reaction only at that temperature. They should work best at 37-400 C.

Background Information
Enzymes are biological catalysts. Enzymes consist of various types of proteins that work to drive the chemical reaction required for a specific action or nutrient. Enzymes can either launch a reaction or speed it up. The chemicals that are transformed with the help of enzymes are called substrates. In the absence of enzymes, these chemicals are called reactants.

Shoumit Sarkar

Enzymes have active sites where they come into contact with particular substrates. The catalytic properties of enzymes are a cyclic process. Once a substrate has come into contact with the active site of an enzyme, it is modified by the enzyme to form the end product. Once the process is complete, the enzyme releases the product and is ready to begin the process with new substrates. Enzymes are never wasted and always recycled.

Amylase is an enzyme that breaks starch down into sugar. Amylase is present in human saliva, where it begins the chemical process of digestion. Foods that contain much starch but little sugar, such as rice and potato, taste slightly sweet as they are chewed because amylase turns some of their starch into sugar in the mouth. The pancreas also makes amylase (alpha amylase) to hydrolyse dietary starch into disaccharides and trisaccharides which are converted by other enzymes to glucose to supply the body with energy. Plants and some bacteria also produce amylase.

Apparatus
3 beakers 6 test tubes Test tube rack Spotting Tile Starch solution Amylase solution Iodine Hot water baths

Shoumit Sarkar

Method

Solution 27 degrees

Place 5ml starch solution in a test tube Place 5ml amylase in another test tube Heat in water bath for 5 minutes o 27, 38, 60 degrees Add amylase to starch o Shake and put back in water bath for another 5 minutes. Place several drops in the white tile Solution 37 degrees Add one drop iodine to each one of them Observe results

Solution 60 degrees

Observations and Conclusions

The solution which had been placed in the 27 degree water bath had changed into a brownish yellow colour when iodine solution was added. This suggests that there was no starch remaining in the solution, and that amylase was functioning. The starch had broken down into protein. The temperature was below optimum temperature so the enzymes were not denatured but they would have taken longer than usual to function. The iodine did not turn blue black suggesting that there was no starch left and all the starch had indeed turned into protein. The solution which had been placed in the 37 degree water bath had changed into a brownish yellow colour also, but it had a little bit of blue in it, showing that there was very little or perhaps no starch was remaining in the solution. The amylase had also functioned. This temperature is very close to the optimum temperature and the rate of reaction of the enzymes would have been the fastest at this temperature. Just like earlier, we can see that the solution had mostly not turned blue black suggesting that the majority of the starch was broken down into proteins. A minority of the starch hadnt turned into protein and this explains the slight blue-black color found in the solution. The solution which had been placed in the 60 degree water bath had changed into a dark brown colour and had heavy shades of blue in it. This tells us that the solution had some starch in it and also suggests that the amylase either did not function properly, or had very little amounts of amylase in it. Compared to earlier, these enzymes were put in a solution with a very high temperature. The enzymes would obviously have been denatured and they would not have functioned. This explains the blue black color. The starch would have obviously not broken down into proteins. This experiment could have been extended by using a wider range or temperatures. Eg. 15-1000