Enzyme Function Lab Summary Assignment

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Feb 20, 2024

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Angel Nguyen BIOLOGY 181 Lab 3, Part 1: Effect of pH and Temperature on α -amylase Activity ENZYME LAB SUMMARY ASSIGNMENT (Please download the file on your desktop, complete, and submit by the due date) INTRODUCTION The purpose of this lab is to become familiar with enzyme function , and to understand how changes in pH and temperature values affect enzymatic activity . In this laboratory, we will investigate the effects of temperature and pH on the activity of α -amylase extracted from the three different organisms (sources) listed below : • Bacillus licheniformis (bacterium), whose α -amylase functions optimally at high temperatures (65-100°C) and pH 6, 8 • Aspergillus oryzae (microscopic fungus), whose α -amylase functions optimally at 37°C and pH 5,6 (acidic) • Pig pancreas, whose α -amylase functions optimally 37˚C, and pH 7 (neutral). OBJECTIVES: • Understand the procedure for identifying the source of the three unknown α - amylase (A, B, or C) • Identify the source of three different α -amylase (A, B, or C) by determining the optimal pH and temperature values of each α -amylase (A, B, or C) • Calculate concentration of maltose (mg/mL) produced by α -amylase A, B, or C at optimal conditions of pH and temperature TASKS: • Identify the source of three different α -amylase (A, B, or C) by determining the optimal pH and temperature values of each α -amylase (A, B, or C) • Analyzing Results and Identifying the Source of Enzyme A, B, and C Question 1 (3 points): What is the source of enzyme A? Why? Is it bacterium Bacillus licheniformis ? Is it the microscopic fungus Aspergillus oryzae ? Is it pig pancreas? To answer the question, match the optimal values of pH and temperature (in pink in the tables) with those of bacterium, fungus, or pig pancreas spelled out in the “Introduction” section The source of Enzyme A is Aspergillus oryzae. Aspergillus oryzae derived Enzyme A as the enzyme made by a particular filamentous fungus. For Enzyme A the optimal pH levels were 5 and 6. The optimal temperature was 37˚C therefore the souce of Enzyme A is Aspergillus oryzae

Question 2 (3 points): What is the source of enzyme B? Why? Is it bacterium Bacillus licheniformis ? Is it the microscopic fungus Aspergillus oryzae ? Is it pig pancreas? To answer the question, match the optimal values of pH and temperature (in pink in the tables) with those of bacterium, fungus, or pig pancreas spelled out in the “Introduction” section. The source in Enzyme B is Bacillus licheniformis as it is referring to a particular enzyme that bacterium produces. A species that is know for this is subtilisin a type of serine protese which is produced by Bacillus licheniformis. The optimal pH for Enzyme B was at 6 of 0.22 and 8 at 0.24 the optimal temperature was at 65-100°C therefore Enyme B is Bacillus licheniformis. Question 3 (3 points): What is the source of enzyme C? Why? Is it bacterium Bacillus licheniformis ? Is it the microscopic fungus Aspergillus oryzae ? Is it pig pancreas? To answer the question, match the optimal values of pH and temperature (in pink in the tables) with those of bacterium, fungus, or pig pancreas spelled out in the “Introduction” section. The source of Enzyme C is a pigs pancreas. The reason is that it is referring to the "pancreatic lipase". In humans, pigs, and other mammals, the pancrease produces the enzyme known as pancreatic lipase. The main objective of that is convert dietary fats also known as lipids into simpler molecules such as fatty acids and glycerol so that the small intestine can absorb easily. In the experiment the optimal pH levels was 7 of 0.41 and the temperature at 37˚C, therefore Enzyme C was pigs pancreas. • Calculate concentration of maltose (mg/mL) produced by α -amylase A, B, or C at optimal conditions of pH and temperature using the maltose standard curve generated in lab 2 Exercise 4 (3 points): Calculate the concentration of maltose of Enzyme A, B, and C, optimal pH and temperature values Table 8. Enzyme A, B, and C production of maltose (mg/mL) at optimal pH and temperature values Enzyme optimal pH and temperature absorbance @ 540nm (abs units) concentration of maltose (mg/mL) Enzyme A optimal pH 0.66 1.89 Enzyme A optimal T 0.65 1.86 Enzyme B optimal pH 0.24 0.69 Enzyme B optimal T 0.95 2.73 Enzyme C optimal pH 0.41 1.18 Enzyme C optimal T 0.27 0.77 Example (Enzyme A optimal pH, 0.66): x (concentration of maltose mg/mL) = = 1.89 mg/mL of maltose Question 5 (3 points): Once you have identified the source of enzyme A, B, and C,

please compare the optimal temperature of each enzyme and answer the following questions: Why enzyme A optimal temperature is 37°C? Why enzyme B optimal temperature is 65°C - 100°C? Why enzyme C optimal temperature is 37°C? Recall that enzymes are adapted at the conditions of pH and temperature of the cell environment where they operate!! Optimal temperature is the temperature at which the enzyme is most active and produces more produces in little time. At high temperatures, the enzymes are denatured, while at low temps the enzymes are not active. The pig pancreatic (Enzyme C) has a maximum temperature of 37°C because the organisms is a warm body/blooded mammal and maintains a core temp of about the same. Same can be said with Enzyme B as Bacillus licheniformis has an optimal temperature at 65°C - 100°C becase this organism is an extremophile it thrives at high temps between 41-122 celcius. As for Enzyme A it also habitats at the temperature of 37 celcius. Thank You for Completing Lab 3, Part 1!

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