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The effect of enzyme concentrations on the reaction time of Urease active meal.

Extracts from this document...

Introduction

Title: The effect of enzyme concentrations on the reaction time of Urease active meal. Aim: My aim of this investigation is to investigate the effect of enzyme Urease concentrations on reaction time. Urease is the enzyme used to breakdown urea into carbon dioxide and ammonia. I have chosen to carry out this investigation because there are many different areas of biology linked with this investigation that are of interest to me. In addition, enzymes are one of the most important and fascinating molecules found in the body and I would enjoy learning more about the fascinating compound for my own pleasure. Introduction: Enzymes are biological organic catalysts. They are specialised globular proteins with a complete tertiary structure that give it the property of being specific for one biochemical reaction that takes place inside every living cell. Many reactions that take place inside living organisms are very slow without the presence of an enzyme. Enzymes act as biological catalysts that allow reactions to take place rapidly in conditions that are found inside living cells without being chemically used up or changed themselves. Each different type of enzyme has a unique three-dimensional structure that enables the enzyme to catalyse one type of reaction. They may affect one particular biochemical reaction strongly but leave a similar reaction unaffected. In an enzyme catalysed reaction, the reactance, also known as the substrate, binds to the enzyme at specific points known as the active site to form an enzyme-substrate complex. The precise shape of the acitive site is important inorder for the substrate to combine to the enzyme as the substrate is the complementary shape to the active site. The diagram below represents this: The overall three - dimensional shape of a polypeptide chain is referred to as the enzymes tertiary structure. In globular proteins, the polypeptide chains are tightly folded to form a spherical shape. Many globular proteins are folded so that their hydrophobic groups are on the inside of the molecule and the hydrophilic groups face outwards, making these proteins suitable in water. ...read more.

Middle

- to contain standard solutions accurately * 200cm3 volumetric flask &stopper (x2) - to contain standard solutions accurately * 1000cm3 volumetric flask &stopper (x1) - to contain standard solutions accurately * Funnel - for accurate manipulation of solutions * Test tubes ( 8 x 12) - reaction vessel * Test tube rack - to hold test tubes * Labels - to avoid any confusion with solution and prevent anomalies in my results * Syringe (x4) - to measure and transfer small amounts of solutions accurately * Stop clock - to measure the time Chemicals: * Distilled water and wash bottle - to rinse out impurities and dilute solutions * 3% Urease solution - source of enzyme * 2% urea solution - source of substrate * pH5 buffer solution - to maintain pH of solution * pH7 buffer solution - to maintain pH of solution * pH7.5 buffer solution - - to maintain pH of solution * Phenolphthalein indicator - to indicate end point of the reaction * Bromothymol Blue indicator - to indicate end point of the reaction Risk assessment Below is a list of all the chemicals I will be using for my experiment and the risks involved with them: * Urea - toxic * Ammonia - highly toxic * pH5 solution - weak alkali * pH7 buffer solution - * pH7.5 buffer solution - * Urease enzyme - * Phenolphthalein indicator - irritant to eyes, skin and respiratory system. * Bromothymol Blue indicator - Safety Extreme care must be taken when handling all the chemicals mentioned above, as they can be hazardous not only to myself but to other people around me. To ensure the safety and minimise risk from the nature of the hazards exposed to others, and myself I will: * Make sure that protective clothing i.e. safety glasses; lab coat and protective gloves are worn throughout the experiment. * Keep hazardous chemicals away from naked flames, as the chemicals I am using are flammable. ...read more.

Conclusion

in minutes, of enzyme concentration using Bromothymol blue indicator Experiment 2 - using Bromothymol blue Test tube number Rough 1 2 3 4 5 6 7 8 9 10 11 12 1 11.33 11.31 1.32 12.06 11.52 11.42 11.43 11.42 11.24 11.53 11.37 11.48 11.36 2 7.06 6.29 6.52 6.23 6.42 6.27 6.16 6.14 6.26 6.34 6.25 6.32 6.46 3 4.32 4.50 4.53 4.53 4.42 4.29 4.38 4.41 4.42 4.16 4.35 4.59 4.51 4 2.43 2.53 2.44 2.39 2.46 3.05 2.43 2.46 2.47 2.58 3.03 3.01 2.58 5 2.15 2.25 2.23 2.22 2.18 2.17 2.19 2.20 2.21 2.23 2.19 2.16 2.21 6 2.08 2.03 1.56 1.53 2.03 1.41 1.56 2.03 2.04 1.52 1.49 2.05 2.06 7 1.23 1.16 1.13 1.14 1.10 1.21 1.15 1.20 1.17 1.18 1.09 1.22 1.23 8 1.02 10.53 0.54 0.49 1.03 1.06 0.48 0.46 0.57 0.53 0.47 0.51 0.55 Analysis Below is a table showing the reaction time converted into seconds. Experiment 1: phenolphthalein Test tube number Rough 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 Experiment 2 - using Bromothymol blue Test tube number Rough 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 Below is a table showing the average of my results in seconds, the reaction time in seconds, and the weighted reaction time. Average = 1 + 2 + 3 + 4 + 5 + 6 + 7 + 8 + 9 + 10 + 11 + 12 12 weighted reaction time = average reaction time (secs) X conversion factor Experiment 1: phenolphthalein Test tube number Reaction time (secs) Conversion factor Weighted reaction time (secs) 1 0.0667 2 0.1333 3 0.2000 4 0.2667 5 0.4000 6 0.5333 7 0.6667 8 1.0000 Experiment 2: Bromothymol blue Test tube number Reaction time (secs) Conversion factor Weighted reaction time (secs) 1 0.0667 2 0.1333 3 0.2000 4 0.2667 5 0.4000 6 0.5333 7 0.6667 8 1.0000 ...read more.

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