BIO 110 Enzyme Action Lab - Part I

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Mars Hill College *

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110

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Biology

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May 2, 2024

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docx

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Lab Enzyme Action - Part I Pre-lab questions 1. Enzymes belong to what class of biologically important molecules? u Enzymes belong to a class of biologically important molecules called proteins. u Amino Acids 2. What is a hydrolytic enzyme? u It is an enzyme that breaks lipids, carbohydrates, proteins, fats, and nucleic acids into their simplest units. u A hydrolytic enzyme is an enzyme that carries out an hydrolysis reaction. 3. What hydrolytic enzyme will be used in this lab? u Enzyme Amylase 4. What is the substrate of this enzyme and how will it be determined in the lab if enzyme activity has occurred? u Starch is the substrate enzyme. It turns dark blue in the presence of iodine as it
reacts with after a break down u to glucose through heating u Starch is the substrate enzyme. It turns dark blue in the presence of iodine as it reacts with after a break down u to glucose through heating u Starch is the substrate enzyme. It turns dark blue in the
presence of iodine as it reacts with after a break down u to glucose through heating u Starch is the substrate enzyme. It turns dark blue in the presence of iodine as it reacts with after a break down u to glucose through heating u Starch is the substrate enzyme. It turns dark blue in the presence of iodine as it reacts with after a break down to glucose through heating. u The substrate of this enzyme is starch amylose which is breaking down. If there is not substrate or starch detected, then the enzyme activity has occurred. Because it is the enzyme job to break down the substrate starch.
And if it is not detected then it means that the enzyme activity has taken place. 5. What type of dilution will you be completing in Part I? u We Will be completing a two fold serial dilution in part 1. Objectives: 1. Define the terms substrate and product of an enzyme-catalyzed reaction. 2. Observe a digestion reaction using amylase and determine when the reaction is complete. 3. Describe how the shape of a protein is influenced by its physical environment. 4. List at least 3 factors in the environment that could affect the activity of an enzyme. 5. Develop a hypothesis regarding the activity of the amylase enzyme with respect to a particular environmental condition. (SLO) 6. Set up a well-controlled experiment to test your hypothesis. (SLO) Introduction The action of enzymes is required in order for chemical reactions to occur at an appreciable rate in cells. In the absence of cellular enzymes, glucose would not be available or usable in fueling reactions, the formation and breakdown of ATP would not occur, and macromolecules would not be formed or recycled to monomers, to name a few important reactions in cells. Simply put, life would not exist. In this lab we will conduct investigations on amylase, a hydrolytic enzyme present in human saliva that normally acts to breakdown the biomolecule starch (the substrate of the reaction) to simpler sugars (the product of the reaction) during the process of food digestion. We will use this enzyme as a model system to examine the effect of environmental changes on amylase activity with the goal of developing a better understanding of the importance of protein structure on the function of an enzyme. Chemical reaction of starch hydrolysis with the release of the disaccharide maltose facilitated by the enzyme amylase:
Part I: Serial dilution to determine optimal amylase activity 1. Watch the video and follow the workflow below for instruction on how to complete a 2-fold serial dilution of the “stock saliva”. Start by making a 1:2 dilution by transferring 2ml of the “stock saliva” to a test tube containing 2ml of H2O. Label this tube 1:2. Continue the serial dilution until you reach a 1:32 dilution (transfer 2 ml of the 1:2 saliva dilution to 2ml of H2O (1:4 dilution), ect).
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