Denise Petite Anne A.

Siojo

September 10, 2013

Enzymes

Abstract

The effect of temperature on catalase and amylase were tested. They functioned well at optimum temperature. The enzyme treated above the optimum temperature denatured and no longer functioned. The enzyme exposed at room temperature worked slowly. Temperature affects the functionality of enzymes. Introduction

Enzymes are biological catalysts that carry out the thousands of chemical reactions that occur in living cells. They are proteins that act as catalysts for biological reactions. Enzymes, like all catalysts, speed up reactions without being used up themselves. They do this by lowering the activation energy of a reaction. All biochemical reactions are catalyzed by enzymes. Since enzymes are proteins, they can be denatured in a variety of ways, so they are most active under mild conditions. Most enzymes work best at optimum temperature and pH. The optimum temperature is usually around body temperature (37C). Substrate concentrate and inhibitors also affect the condition of the enzyme. Enzymes are also very specific they only act on one substrate or one class of related substrate molecules. The reason for this is that the active site of the enzyme is complementary to the shape and polarity of the substrate. Typically, only one kind of substrate will fit into the active site. Above or below the optimum conditions, the enzymes the may work slowly. Some may be denatured and cannot fully function. Catalase and amylase are examples of an enzyme. The latter is responsible for hydrolyzing starch. It can be found in human saliva, where it begins the chemical process of digestion.In its presence, a sample of starch will be hydrolyzed to shorter polysaccharides, dextrins, maltose, and glucose. Catalase on the other hand accelerates the breakdown of hydrogen peroxide (a common end product of oxidative metabolism) into water and oxygen. In this experiment, the effect of temperature on catalase and amylase is tested.

Methods and Materials

Preparation and Action of Catalase

A small potato was pared and chopped into a fine pulp. The pulp was mixed with 50mL water and was set aside for 15 minutes. It was strained through a piece of cheesecloth. Then it was filtered. 15 drops of the filtrate was placed in a test tube and was boiled in a water bath. It cooled then added with 5 drops of 5% hydrogen peroxide. The procedure was repeated using unboiled filtrate instead.

Action of Amylase

In different test tubes, 15 drops of starch solution was place on each. They were marked as test tube 1, 2, and 3 respectively. Test tube 1 was heated to boiling, added with 5 drops of saliva, and the boiling continued for one more minute then it was cooled. On test tube 1, an mL of saliva was added on it. It was placed in a beaker preheated to 37oC for 30 minutes. And lastly test tube 3 was added with 1 mL saliva and was kept on room temperature. All three test tubes were added with Fehlings reagent with iodine.

Results and Discussion

In the unboiled filtrate, bubbles formed upon adding hydrogen peroxide. This indicates the presence of oxygen. Catalase breaks down hydrogen peroxide to oxygen and water. The catalase on the unboiled filtrate hydrolyzed the hydrogen peroxide. In the experiment where the filtrate was boiled, when it was added with hydrogen peroxide it didnt form bubbles. This is because the catalase didnt function. The temperature was already above the optimum temperature. Unfolding of the protein happened thus the catalase in the boiled filtrate denatured. Denaturation of protein also happened in the experiment using amylase. Amylase breaks down starch into simpler sugars. In test tube 1, upon adding Fehlings reagen with iodine the solution turned blue. This indicates the presence of starch. Starch
reacts with iodine to produce the blue starch-iodine complex. Amylase didnt function

because it was heated to boiling causing the enzyme to be denatured. In test tube 2, where it was preheated to 37OC, when added with Fehlings reagent, the solution turned blue-green with red precipitate. This indicates a positive result with Fehlings reagent. Starch had been hydrolyzed to simple sugars. The amylase in test tube 2 worked well because it was treated with the optimum temperature. The result in test tube 3, was also positive with Fehlings reagent. The color of the solution seen was purple. The rate of reaction is slow when the temperature is below the optimum temperature. A little amount of starch may still be present.

Conclusion The speed and functionality of chemical reactions is affected by temperature. Enzymes work best at an optimum temperature. Above the optimum temperature, the enzymes are denatured. Bonds holding the structure together will be broken and the active site loses its shape and will no longer work. At lower temperature, the activity of the enzyme is also low.

References

Effect of Catalase on Hydrogen Peroxide. Retrieved on 7 September 2013 from <http://www.sciencegeek.net/Biology/biopdfs/Lab_Catalase.pdf>

Enzymes.

Retrieved

on

September

2013

from

<http://courses.chem.psu.edu/chem112/materials/enzymes.pdf>