Studying enzymes under different temperatures

The experiment was directed to research the effects temperature has on enzymes and the reaction times of breaking down starch at different temperatures. The role of the enzyme can be disturbed by temperature, PH, salinity, enzyme and substrate concentration, and the presence of inhibitors or activators. There were two enzymes brought into the experiment conducted, one was Bacterial Amylase and the other was Fungal Amylase. Eight test tubes for each amylase (Fungal and Bacterial) are needed, four containing starch and for containing amylase. Each test tube with starch and with amylase is placed into designated water baths temperatures of 0, 25, 65, and 85 degrees Celsius and left for a certain amount of time to see the reactions time to break down starch when combined. The time the test tubes are placed in the water baths are 0, 2, 4, 6, 8, and 10 minutes. At each minute a pipette is placed in the test tube and 2-3 drops of the solution of the test tube is placed on the spot plate slot designated at each time mixed with 2-3 drops of Iodine. Depending on the color at each minute is the effect the temperature had on the reaction time of the breaking down of starch. Each result should be recorded to combine the results of both Bacterial and Fungal Amylase. This would allow for conclusion of the experiment.

INTRODUCTION

Cells require enzymes to survive and function. Enzymes are catalysts, a catalyst is a substance that increases the rate of a chemical reaction without itself undergoing any permanent chemical change. The enzyme lab is to see how the rate of enzyme activity is affected due to the changes in temperature, salinity, and pH levels. If the temperature is too high or too low the enzyme will start to denature. Denaturing means that the enzyme will start to unfold or lose its shape and therefore stop working because it can no longer bind to its active site. In this lab the study of specifically Bacillus lichenformis and Aspergillus oryzae, which are enzymes, will be studied at different temperatures. By studying the enzymes under different temperatures combined with starch will demonstrate at which temperature the enzyme works best. When starch is broken down the product will be yellowish coloring. When the starch combined with the enzyme is not broken down the product will be black/blue. Bacillus lichenformis is commonly found in soils, plant material, or bird feathers (ducks and sparrows) around the world. It is used in the industry to secrete proteins from the enzyme creating antibiotics. The enzyme thrives in temperatures of 50 degrees Celsius or higher. It can survive in bad environments by turning into a spore (Mai, 2011). The other enzyme being used for starch breakdown is Aspergillus Oryzae. This enzyme is commonly found in the ground as it is a type of mold. This tends to thrive in temperatures of 30 to 45 degrees Celsius. It is used in the industry but more commonly towards the Asian culture for food fermentation (Nanavatv, 2015). In the lab 4 tubes for each enzyme will be put into water baths of 0,25,65, and 85 degrees Celsius and then be combined with the starch and some iodine to see how the enzyme reacts to each temperature.

METHODS

Procedure

In this experiment two spot plates were used, one for each enzyme and labeled on the y-axis with time and the x-axis with temperature of water bath it was placed in. The times used for the experiment were intervals of two from zero to ten. The temperatures used were 0,25,65, and 85. Four of the test tube is labeled by Fungal or Bacterial starch and the other four for each group are labeled Fungal or Bacterial amylase. For each starch test tube a solution of 5mL containing 1.5% starch was placed into each set. The amylase test tubes will not be containing starch. When all done place each test tube into their pertaining water bath temperatures. To not get confused it is recommended to do the experiment first with Bacterial tubes, the next time around with the Fungal tubes. To start off the experiment the test tubes must acclimate to their water bath temperature so an interval of 5 minutes should be provided before starting experiment. Each time the stopwatch beeps 2-3 drops of iodine should be placed in spot plate spot which corresponds to the tube and water bath temperature and a pipette should be used to transfer some solution to the spot plate without removing the test tube from its water bath. This should be done each time when the stop watch beeps until all spot plate spaces are full. A different pipette should be used for each solution. After the first round the starch solution should be mixed with the amylase solution test tube and continue the process along. Record each change seen on spot plate for experiment results at the end of each transfer from test tube to spot plate. (Alberte, Jose, Pitzer, Calero, 2012)

Source: Essay UK - https://www.essay.uk.com/essays/science/studying-enzymes-under-different-temperatures/


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