...LAB #3: An Enzyme in Plant and Animal Tissues BACKGROUND INFORMATION Liver and other living tissues contain the enzyme catalase. This enzyme breaks down hydrogen peroxide (H2O2), which is a harmful by-product of cellular respiration if it builds up in cells. In this lab, you will perform reactions with the enzyme catalase. OBJECTIVES: 1. Investigate the enzyme catalase in various tissues. 2. Measure the effect of changes in temperature on reaction rates of an enzyme-catalyzed reaction in a controlled experiment. INTRODUCTION: What would happen to your cells if they made a poisonous chemical? You might think that they would die. In fact, your cells are always making poisonous chemicals. They do not die because your cells use enzymes to break down these poisonous chemicals into harmless substances. Enzymes are proteins that speed up the rate of reactions that would otherwise happen more slowly. The enzyme is not altered by the reaction. You have hundreds of different enzymes in each of your cells. Each of these enzymes is responsible for one particular reaction that occurs in the cell. In this lab, you will study an enzyme that is found in the cells of many living tissues. The name of the enzyme is catalase (KAT-uh-LAYSS); it speed up a reaction that breaks down hydrogen peroxide, a toxic chemical, into 2 harmless substances--water and oxygen. The reaction is as follows: 2H2O2 à 2H2O + O2 This reaction is important to cells because hydrogen peroxide...
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...Christian Rodriguez Mrs. Gentry AP Biology September 27, 2015 Lab Abstract Enzymes are proteins produced by living cells. In an enzyme, the substrates bind with the active site of an enzyme, breaking them down into other molecules. Enzymes acts as catalyst which lower the reaction energy and allows it to take place in less time and at lower temperatures. Enzymes can be used in a reaction without actually being used up; therefore, they can be used reversibly. Temperature, salt concentration, substrate concentration, enzyme concentration, and pH can affect enzyme reactions. If the pH is too basic, the enzyme gains (H+) and denaturalizes; likewise if the pH is too acidic, then the enzyme loses (H+) and becomes denaturalized. A major benefit...
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...Enzyme Lab INTRODUCTION: What would happen to your cells if they made a poisonous chemical? You might think that they would die. In fact, your cells are always making poisonous chemicals. They do not die because your cells use enzymes to break down these poisonous chemicals into harmless substances. Enzymes are proteins that speed up the rate of reactions that would otherwise happen more slowly. The enzyme is not altered by the reaction. You have hundreds of different enzymes in each of your cells. Each of these enzymes is responsible for one particular reaction that occurs in the cell. In this lab, you will study an enzyme that is found in the cells of many living tissues. The name of the enzyme is catalase (KAT-uh-LAYSS); it speeds up a reaction which breaks down hydrogen peroxide, a toxic chemical, into 2 harmless substances--water and oxygen. The reaction is: 2 H2O2 ----> 2 H2O + O2 This reaction is important to cells because hydrogen peroxide (H2O2) is produced as a byproduct of many normal cellular reactions. If the cells did not break down the hydrogen peroxide, they would be poisoned and die. In this lab, you will study the catalase found in liver cells. You will be using chicken or beef liver. It might seem strange to use dead cells to study the function of enzymes. This is possible because when a cell dies, the enzymes remain intact and active for several weeks, as long as the tissue is kept refrigerated. MATERIALS: 1molar HCl solution 1molar NaOH...
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...Title: What is the effect of temperature on Enzyme activity? Purpose: Enzymes are proteins that speed up chemical reactions in cells. They break down molecules called substrates. Each enzymes have only one substrate that breaks down. Enzymes are produced in the cells of the body and affect the rate of almost all the chemical reactions which take place in living organisms. The rate of enzymes activity is influenced by temperature, pH, and substrate concentration. The purpose of this lab was to determine the affects of enzyme activity under specific temperature changes, pH values and substrate concentration. Since heat increases the rate of most chemical reactions, the addition of heat causes faster molecular movement. Most enzymes active in living tissue becomes denatured, their secondary or tertiary protein structure breaks down, at the temperature above 40 degrees C. In the effects of pH, it is expected that the changes in pH would have an effect on the action of enzymes. Lemon juice helps keep the apple from growing, because its full of ascorbic acid (Vitamin C) and it has a low acidic pH level. But extreme high levels of low pH values can result in a complete loss of enzymes activity thus leaving the apple to brown. The effects of substrate concentration, an enzyme substrate complex is formed when a substrate fits into active of an enzyme. The velocity, the rate of speed, at which the enzymes works will increase until it reaches a maximum. If the substrate...
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...Enzymes activity plays an important role on living organisms. Its activity can be affected by different factors, including environmental and molecular elements. Enzyme inhibition by some molecules can be competitive or noncompetitive according to the binding mechanism. The aim of this study was to determine the type of inhibition occurred by the phenylthiourea (PTU) on the catechol oxidase enzyme. Different substrate concentrations, 0.5, 1, and 1.5 mL, and presence (1mL) and absent of PTU was analyzed. Products were measured at 380 nm. Results showed product only when PTU was not present, except for in one tube. A PTU noncompetitive inhibition is proposed based on the inability to obtained product even when the substrate concentration is increased....
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...Biotechnology is a field of applied biology that involves the use of living organisms and bioprocesses in engineering, technology, medicine and other fields requiring bio products. Father of the term biotechnology is ‘Karl Ereky’ and Father of Biotechnology was ‘Louis Pasteur’. DNA, or deoxyribonucleic acid, is the hereditary material in humans and almost all other organisms. Nearly every cell in a person’s body has the same DNA. Ribonucleic acid or RNA, is one of the three major macromolecules (along with DNA and proteins) that are essential for all known forms of life. A pipette (also called a pipet, pipettor or chemical dropper) is a laboratory instrument used to transport a measured volume of liquid. Pipettes come in several designs for various purposes with differing levels of accuracy and precision, from single piece glass pipettes to more complex adjustable or electronic pipettes. Lab dish washing Cleaning laboratory glassware isn't as simple as washing the dishes. Here's how to wash your glassware so that you won't ruin your chemical solution or laboratory experiment. You can rinse the glassware with the proper solvent, then finish up with a couple of rinses with distilled water, followed by final rinses with deionized water Water Soluble Solutions (e.g., sodium chloride or sucrose solutions) Rinse 3-4 times with deionized water then put the glassware away. Water Insoluble Solutions (e...
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...What is the function of enzymes in a living system? Enzymes speed up metabolic reactions necessary for life. Without them certain vital processes would not take place and the body would be unable to function. Difference enzymes work better under different conditions. Where in a human body might it be beneficial to have enzymes work in very acidic environments? In areas, like the stomach, that have a pH of two would benefit by having enzymes that function well in acidic environments. An example of such an enzyme is pepsin. There is a large amount of catalase found in a human liver. Does the liver break down more hydrogen peroxide in the summer or winter? Explain your answer. More hydrogen peroxide will be broken down in the summer compared to the winter because higher body temperatures equals more enzyme activity. Many enzymes end with “ase”. Come up with your own enzyme, then name and explain what this enzyme does. Draw the enzyme and the substrate in the space provided below along with the enzyme-substrate complex. My enzyme would be olestrase. It would break down the lipid olestra and make it usable for the human body. Recent advances have allowed humans to mass-produce certain enzymes. Research one such enzyme and explain how this enzyme has been used to benefit society. Coenzyme Q10 (ubiquinone) is a naturally occurring substance which has properties potentially beneficial for preventing cellular damage during myocardial ischemia and reperfusion. It plays...
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...Introduction Enzymes are protein molecules that catalyze chemical reactions in all living organisms. Enzymes allow living organisms to carry out complex chemical activities at low temperatures, but can’t cause a reaction that hasn’t occurred in their absence. Also, enzymes are thought to speed up reactions by bringing reacting molecules together to increase the chances that a reaction will occur (Worthington Biomedical Corporation, 2015). Each enzyme has a specific active site where the substrates attach. Many factors can affect enzyme activity such as temperature, pH, and the presence of inhibitors (John W. Kimball, 2014). The purpose of this lab was to examine factors affecting the enzyme function of peroxidase. In the 19th century French chemist Louis Jacques discovered catalysts. Catalysts are substances that enable a chemical reaction without participating in it, which led to specifically peroxidases. The structure of peroxidase is a very large enzymatic protein, and has complex molecules with complicated shapes involving multiple folding’s. The activity of peroxidase is dependent on pH. It exhibits maximum activity at a pH between 6.5 and 7.0. The activity of the enzyme is reduced when pH levels are increased. Peroxidase promotes the oxidation of various compounds naturally of peroxides, where hydrogen peroxide is reduced to form water (Wikimedia Foundation, 2015). Also peroxidases break compounds down into harmless substances by adding donor molecules. During this lab, the donor...
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...Factors Affecting Enzyme Activity Analysis of Enzyme Activity: Catalase and Tyrosinase Introduction to Neurotransmitters: Acetylcholinesterase Abstract: A series of three labs were combined to observe the effects of some common biological enzymes: Catalase, Tyrosinase, and Acetylcholinesterase (AChE). Enzymes are catalytic proteins, that when present in a chemical reaction, are able to lower the action potential needed to create the reaction without being destroyed or altered themselves in the process. In Part A, my hypothesis stated that when Catalase is combined with H2O2 the rate of conversion to water and oxygen gas should double when 5-10 drops of enzyme is added and quadruple when 10-20 drops are added. In Part B, my hypothesis stated that increases in enzyme concentration or buffer pH the substrate of the final product will yield increased substrate, also, if the substrate concentration is increased then the enzyme will be less diluted, the buffer pH will increase, or there will be a temperature increase. In Part C, my hypothesis stated that tacrine will have an inhibitory effect on AChE, and that those effects will increase as the level of concentration increases. In all three labs I postulated that increases in temperature and concentration levels and would increase the rates and decrease time to form chemical reactions. We setup each lab with a series of increased concentrations and a control trial using DiH2O. We observed the results using the...
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...effect of temperature, pH, and substrate concentration on Enzyme activity? Purpose: Enzymes are proteins that speed up chemical reactions in cells. They break down molecules called substrates. Each enzymes have only one substrate that breaks down. Enzymes are produced in the cells of the body and affect the rate of almost all the chemical reactions which take place in living organisms. The rate of enzymes activity is influenced by temperature, pH, and substrate concentration. The purpose of this lab was to determine the affects of enzyme activity under specific temperature changes, pH values and substrate concentration. Since heat increases the rate of most chemical reactions, the addition of heat causes faster molecular movement. Most enzymes active in living tissue becomes denatured, their secondary or tertiary protein structure breaks down, at the temperature above 40 degrees C. In the effects of pH, it is expected that the changes in pH would have an effect on the action of enzymes. Lemon juice helps keep the apple from growing, because its full of ascorbic acid (Vitamin C) and it has a low acidic pH level. But extreme high levels of low pH values can result in a complete loss of enzymes activity thus leaving the apple to brown. The effects of substrate concentration, an enzyme substrate complex is formed when a substrate fits into active of an enzyme. The velocity, the rate of speed, at which the enzymes works will increase until it reaches a maximum. If the...
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...non-reproductive cloning Reproductive Cloning: production of offspring which are genetically identical to the mother or the other offspring Non- Reproductive Cloning: Use of stem cells in order to generate replacement cells, tissues and organs which may be used to treat particular diseases 2.2.2- Describe the production of natural clones in plants using the example of vegetative propagation in elm trees Vegetative Propagation: The production of structures in an organism that can grow into new individual organisms These offspring contain the same genetic information as the parent so they are clones of the parent. In Elm Trees: The elm tree is adapted to reproduce asexually following damage to the parent plant. This happens by separation of some body part of the plant body and its development into a new plant. In an elm tree this occurs by: Root Suckers/ Basal Sprouts are removed from a tree in autumn and are grown in a nursery bed. They appear within 2 months of destruction. The suckers grow from meristem tissue in the trunk which is close to the ground (area of least damage) 2.2.3 – Describe the production of artificial clones of plants from tissue culture Tissue Culture: Large scale cloning 1- A small piece of tissue from the plant is taken to be cloned. It is called an explant 2- In aseptic condition, the explant is placed on a nutrient growth medium which contains sucrose, nutrients and growth hormones 3- Undifferentiated cells...
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...Studies VII Practical 9 Cell Biology Studies IX Practical 10 Cell Biology Studies X - Experiment Description Page Writing of Lab Reports Identification of Biomolecules 5 13 Identification of Unknown Carbohydrate Solutions and Investigation of Action of Saliva and HCl in Carbohydrate Solution at Two Different Temperatures Investigation of the Effects of Catalase Concentration on Hydrogen Peroxide Decomposition 20 Synthesis of Starch Using an Enzyme Extracted from Potato Tuber Investigation of the Effects of Different Catalytic Conditions on Hydrogen Peroxide Decomposition Microscopy 27 Practical 6 Cell studies II Practical 7 Cell studies III Extraction of Cell Organelles by Cell Fractionation Determination of Solute Potential of Potato Cell Sap 47 Practical 8 Cell studies IV Effects of Different Treatments on Stained Potato Cells 64 Practical 9 Energetics I Respiration of Germinating Beans 67 Microscopic Examination of Cells at Various Stages of Plant Mitosis and Meiosis DNA, Mitosis and Meiosis Modelling 71 Respiration of Yeast 93 Practical 3 Enzyme studies I (Experiment 1) Optional: Practical 3 Enzyme studies I (Experiment 2) Practical 4 Enzyme studies II Practical 5 Cell studies I - - Practical 10 Energetics II Lab manual version 6_201505 FHSB1214 Biology I & FHSC1214 Fundamentals of Cell...
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...Studies VII Practical 9 Cell Biology Studies IX Practical 10 Cell Biology Studies X - Experiment Description Page Writing of Lab Reports Identification of Biomolecules 5 13 Identification of Unknown Carbohydrate Solutions and Investigation of Action of Saliva and HCl in Carbohydrate Solution at Two Different Temperatures Investigation of the Effects of Catalase Concentration on Hydrogen Peroxide Decomposition 20 Synthesis of Starch Using an Enzyme Extracted from Potato Tuber Investigation of the Effects of Different Catalytic Conditions on Hydrogen Peroxide Decomposition Microscopy 27 Practical 6 Cell studies II Practical 7 Cell studies III Extraction of Cell Organelles by Cell Fractionation Determination of Solute Potential of Potato Cell Sap 47 Practical 8 Cell studies IV Effects of Different Treatments on Stained Potato Cells 64 Practical 9 Energetics I Respiration of Germinating Beans 67 Microscopic Examination of Cells at Various Stages of Plant Mitosis and Meiosis DNA, Mitosis and Meiosis Modelling 71 Respiration of Yeast 93 Practical 3 Enzyme studies I (Experiment 1) Optional: Practical 3 Enzyme studies I (Experiment 2) Practical 4 Enzyme studies II Practical 5 Cell studies I - - Practical 10 Energetics II Lab manual version 6_201505 FHSB1214 Biology I & FHSC1214 Fundamentals of Cell...
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...BIOL 3380 Name:_____________________________________ Circle Session: T-PM W-AM W-PM R-AM R-PM F-AM F-PM Experiment 9 – Pre-lab Homework Enzyme Kinetics of LDH This pre-lab homework assignment is due at the beginning of your lab session. You are provided with the following portion of a protocol: • Determine concentration of enzyme stock solution, if unknown, by taking an A280 nm reading of a 1:100 dilution (in water). Use a total volume of 1 ml in the cuvette. • Dilute some of the enzyme stock with buffer A to make a 4 mg/ml solution. • Serially dilute the 4 mg/ml solution with buffer A to make working solutions of 400 µg/ml and 40 µg/ml. • Prepare 30 µl of each working solution for every sample The PI of the lab gives you a tube of enzyme and tells you the following before disappearing into the office to write more grant proposals: ➢ There is 50 µl of enzyme stock solution. The enzyme is expensive to purify, so follow the protocol exactly, using as little of the stock solution as possible. ➢ The concentration of the stock solution is currently not known, but a 1 mg/ml concentration of the pure enzyme has an A280 nm of 2.0. ➢ You’ll be performing the assay on 12 samples. ➢ Make enough of each working solution so that you have at least 400 ul to work with when you do the assay (to cover any waste and/or inefficiencies in pippetting). Using the spectrophotometer to read the absorbance at 280 nm, you get...
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...INSTRUCTOR GUIDE Human Anatomy & Physiology Laboratory Manual MAIN VERSION, Eighth Edition Update CAT VERSION, Ninth Edition Update FETAL PIG VERSION, Ninth Edition Update ELAINE N. MARIEB, R.N., Ph.D Holyoke Community College SUSAN T. BAXLEY, M.A. Troy University, Montgomery Campus NANCY G. KINCAID, Ph.D Troy University, Montgomery Campus PhysioEx™ Exercises authored by Peter Z. Zao, North Idaho College Timothy Stabler, Indiana University Northwest Lori Smith, American River College Greta Peterson, Middlesex Community College Andrew Lokuta, University of Wisconsin—Madison San Francisco • Boston • New York Cape Town • Hong Kong • London • Madrid • Mexico City Montreal • Munich • Paris • Singapore • Sydney • Tokyo • Toronto Editor-in-Chief: Serina Beauparlant Project Editor: Sabrina Larson PhysioEx Project Editor: Erik Fortier Editorial Assistant: Nicole Graziano Managing Editor: Wendy Earl Production Editor: Leslie Austin Composition: Cecelia G. Morales Cover Design: Riezebos Holzbaur Design Group Senior Manufacturing Buyer: Stacey Weinberger Marketing Manager: Gordon Lee Copyright © 2009 Pearson Education, Inc., publishing as Pearson Benjamin Cummings, 1301 Sansome St., San Francisco, CA 94111. All rights reserved. Manufactured in the United States of America. This publication is protected by Copyright and permission should be obtained from the publisher prior to any prohibited reproduction, storage in a retrieval system, or transmission in any form or by any means...
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