A Report on Enzyme Reaction in a High Temperature Environment

Introduction

Enzyme’s are a extremely important part of chemical reactions not only within us, but all biological creatures. Enzyme’s are used to facilitate and accelerate the chemical reactions that are necessary for not only for us humans to survive, but other creatures like animals and plants to carry out the necessary reactions to produce the materials needed for cellular activity to take place. In order for a accelerated reaction to occur, a substrate will bind to the active site of the enzyme which essentially acts to “accelerate” the reaction speed of an already occurring chemical reaction (George). Without these enzyme’s in the world to accelerate and move along otherwise slow chemical reactions, biological life here on earth would cease to exist. They are key to making reactions take place at speed that are required to sustain life. These enzymes are unique because they do not just disappear once they are used to accelerate a reaction. They allow a reaction to occur at a quicker rate without undergoing permanent alteration while still allowing the reaction to occur at a much faster rate (Tabatabai). This means that once an enzyme is present in a system in order to accelerate a chemical reaction, it can not be used up and will not dissipate unless it comes into the presence of a chemical such as an inhibitor which would therefore slow down the reaction rate between the enzyme and the catalyst. These enzyme inhibitors work to essentially slow the rate at which an enzyme and substrate can bond at the active site. There are several kinds of enzyme inhibitors, nonspecific simply work to slow down any enzyme enhanced reaction, specific inhibitors work only to slow the work of a single type of enzyme or to slow a certain chemical reaction (Enzyme Inhibitors 2015). These inhibitors are often found in poisons and other toxic substances that can kill us and other biological creatures as they are able to slow or bring to a stop chemical reactions that are vital for biological life to sustain itself. Enzymes are very sensitive to environmental conditions and work best under certain circumstances. Overall temperature will accelerate a reaction until a certain point where the reaction will then fall off. When temperature becomes to hot, the enzyme can denature and thus reduce its potency which will slow down any further reaction (Introduction to Enzymes). Studying and understanding enzymes and the processes through which they act, is the first step to understanding how us and other animals are able to survive and thrive on this earth.

Hypothesis:

Through experimentation, I expect the enzyme reaction to work the best in a higher temperature environment, with a neutral pH and balanced amount of enzyme inhibitors to maximize the reaction between the enzyme and the substrate.

Methods and Materials:

There are several different types of experiments that will be conducted over the course of this lab. With each experiment, there will be specific variables that will be changed in order to test the reactions associated with enzymes and substrates at different pH levels, temperature, inhibitors, and concentrations of both substrate and enzyme. In order to accurately test the amount of change in color and therefore the amount of reaction of the substrate and enzyme, a spectrometer machine will be used in order to accurately measure and numerically measure the amount of chance in the cold row the sample. For the first test which will be the controlled reaction test, simply water and substate will be added to different test tubes and results recorded. For the second test which is the control, one of the reactants will be missing while the other conditions will remain the same in order to provide a point of comparison for the future tests to see the difference that changes or variables will have on the reaction. For the third experiment, the specificity of the substate to the enzyme was tested in order to show that the enzyme is designed to react only with a certain enzyme and will not react freely in the presence of any enzyme. The next several tests would test further variables when it comes to enzyme reactions. First the temperature of the enzyme and substrate was adjusted and tested at different temperatures to see at which temperature the enzyme and substrate would work the best at. The next experiment focused on changing the concentration of the enzyme concentration to attempt to find the correct ratio of enzyme to substrate formula to establish what will be the most effective amount. The same would be conducted for the next test but with a varying amount of substrate in this test to show the most important part of and enzyme accelerated reaction. For the final test, inhibitors were added which are used to slow the reaction and their affect on the reaction was recorded in order to learn more about their affects in the reaction process.

Results:

Tube Contents of Tube Initial Color End Color (at 5 min) A400 (at 5 min)

1 10 drop of enzyme

10 drop of substrate yellow tint more yellow tint 0.174

2 10 drop substrate clear clear 0.015

3 10 drop enzyme

10 drop sucrose clear clear 0.005

Table 1: Characteristics of the Enzyme Reactions

Table 2: Effects of Temperature on Enzyme Activity

Tube Temperature (C) Initial Color End Color (at 5 min) A400 (At 5 min)

1 0 yellow tint clear yellow 0.225

2 20 slight yellow tint yellow tint 0.598

3 34 yellow tint dark clear yellow 0.594

4 60 yellow tint dark clear yellow 0.135

5 100 clear practically clear 0.590

Table 3: Effect of Enzyme Concentration on Enzyme Activity

Tube Contents of Tube Initial Color End Color (at 3 min) A400 (at 3 min)

1 3 drops of enzyme clear yellow tint 0.76

2 9 drops of enzyme yellow tint dark yellow 0.305

3 27 drops of enzyme yellow tint dark yellow 0.412

Table 4: Effect of Varying the Substrate Concentration

Tube Contents of Tube Initial Color End Color A400 (at 5 min)

1 3 drop of substrate

10 drop of enzyme slightly yellow darker yellow tint 0.207

2 9 drop of substrate

10 drop of enzyme slightly yellow darker yellow tint 0.211

3 27 drop substrate

10 drop of enzyme slightly yellow darker yellow tint 0.206

Table 5: Effect of pH on Enzyme Activity

Tube pH of Tube Initial Color End Color (at 5 min) A400 (at 5 min)

1 3 clear clear 0.01

2 5 clear light yellow 0.78

3 7 light yellow darker yellow 0.261

4 9 clear clear 0.042

5 11 clear clear 0.063

Table 6: Effect of Inhibitors on Enzyme Activity

Tube Contents of Tube Initial Color End Color (at 5 min) A400 (at 5 min)

1 1 drop pnentithiorn clear clear 0.023

2 20 drop pnentithiorn clear clear 0.010

3 20 drop pnentithiorn clear clear 0.014

4 1 drop tyrosine light yellow light yellow 0.342

5 10 drop tyrosine light yellow light yellow 0.596

6 20 drop tyrosine light yellow light yellow 0.285

7 DI H20 + sol light yellow light yellow 0.3

Discussion:

The results of this experiment are in line with what we expected before the start of this experiment. Enzyme reactions are very much affected by the conditions of the surrounding environment. Enzymes work best in certain ranges of pH, temperature, concentration and other environmental factors. Change these factors too much and the enzyme’s either accelerate or could possibly completely use their ability. With our results in line for the most part with expected results, I believe our experiment was overall a success and can be marked up as one.

Conclusion:

This lab proved much of what I had already learned and understood about enzymes and how they operate to accelerate and enhance chemical and biological reactions. Enzyme’s have very specific conditions under which they work the best and through the many experiments conducted in this examination of enzymes. For one, a balanced pH is necessary for quick processing of substrate by an enzyme. At the same time it is also important that there is more enzyme then substrate in the mixture in order to maximize the reaction speed by allowing there to be plenty of enzyme to process through the substrate at a quick rate. At the same time, the speed of the reaction is also affected by temperature, and through our experimentation, 20 degrees Centigrade is the butter zone for substrate-enzyme reaction. A higher or lower temperature will affect the speed and amount of reaction in either a negative or positive manner.