Lab Section 13154
Title: The Temperature Effect on the Interactions Between Porcine Pancreas
Alpha-Amylase and 1% Starch Solution
Enzymes are virtually the additional power source that allows the continuation of life. The incredible powers of enzymes are conducted by complex protein molecules that are biological catalysts (Ophardt, 2003). Enzymes can vary vastly in the biological world by being categorized for performing different functions. Specifically, the porcine pancreas alpha-amylase (PPA) enzyme performs a complex process that catalyzes the breakdown of starch into sugars by breaking the starch chains into two or three glucose units (Goodsell, 2006).
Since porcine pancreas alpha-amylase breaks down starch, it is widely present in plants, mammalian tissues, and microorganisms (Buisson, Duee, Haser, and Payan,
1987). PPA is an extremely complex enzyme that is composed of minute chemical performances conducted within the different structures of enzymes. The different components that essentially construct PPA begin with a single polypeptide chain of
496 residues that contains five disulfide bridges and two free thiol groups (Buisson,
Duee, Haser, and Payan, 1987). With 496 residues compact into an infinitesimal enzyme, the functions of these residues may vary from binding and/or catalyzation process and calcium ion interactions (Buisson, Duee, Haser, and Payan, 1987).
Although enzymes have hundreds of residues cohesively working together to perform its functions, specific conditions can affect its performance. Temperature is a major influence to the activities of enzymes. An increase in temperature causes all 3 or
Lab Section 13154
4 structures to denature, which can permanently terminate enzyme activities (Clark,
2007). In low temperature environments, the process of enzyme activity will dramatically slow down and if presented below -65 degrees Celsius, the group of atoms and functions within the protein will cease (More, Daneil, and Petach, 1995). Since both high and low temperature changes are detrimental to enzyme activities, the optimum temperature specifically for porcine pancreas alpha-amylase, is 37 degrees
Celsius (Rudeekulthamrong and Kaulpiboon, 2012).
Since it is hypothesized that the increase or decrease in temperature around the porcine pancreas alpha-amylase’s environment will detrimentally effect its original functions, an experiment will be conducted to formally assist the expectations. It is predicted that if the porcine pancreas alpha-amylase were placed into 80, 37, 22 and 4 degrees Celsius, then all of the porcine pancreas alpha-amylase performances will be hindered or terminated except for the optimum temperature of 37 degrees Celsius that will suffice.
Materials and Methods:
A set of eight standard test tubes were prepared and labeled 1-4 for the first four tubes and 1A-4A for the last four tubes. Using a clean 5-mL calibrated pipette, 2 mL of 1% w/v starch solution from the Carolina Biological Supply Company were inserted into the first four test tubes. With a new and clean 5-mL calibrated pipette, each of the four test tubes were mixed with an additional 4 mL of distilled water and 1 mL of WWR International/Micro Essential Laboratories 6.8 buffer.
Lab Section 13154
As for the second set of the four test tubes that were labeled 1A-4A, each were inserted 1 mL of 1% w/v solution alpha-amylase from porcine pancreas (Type VI-B, greater than or equal to 10 units/mg solid) that was obtained from Sigma-Aldrich with a clean 1-mL calibrated pipette. After the mixtures, test tubes 1 and 1A were placed in an 80 degrees Celsius water bath. Test tube 2 and 2A were placed in a warm 37 degrees Celsius water bath while test tube 3 and 3A were placed on a test tube rack at a 22 degrees Celsius room temperature. The last two test tubes were placed in a 0 degrees Celsius beaker of crushed ice. All of the test tubes were placed in the different