Cellular Metabolism

In this experiment, the naturally occurring peroxidase is extracted from homogenized turnip (Brassica rapa) pulp (Coleman 2016). Its role in the environment is to remove toxic hydrogen peroxide during metabolic processes where oxygen is used (Coleman 2016). The goal of this experiment is to evaluate the change of absorbency of turnip peroxidase within a metabolic reaction utilizing oxygen. Any change noted is indicative of the peroxidase removing hydrogen peroxide. Within this experiment, the extract will be prepared, the amount of enzyme will be standardized, and the effect of changing the optimal conditions will be observed. If the enzyme concentration is increased then the rate of the reaction decrease. If the pH of solutions used is increased

The type of peroxidase is used is called turnip peroxidase. Turnip peroxidase is made up of Guaiacol and hydrogen peroxide. The reactants to the product are turnip peroxidase or called tertraguaiacol and water. The color of the react is brown. In the experiment was conducted there were baseline experiment, temperature, pH, 10X substrate, Inhibitor, and half the amount of enzyme.

The experiments were performed in the science lab 1.226 at the University of Texas Rio Grande Valley, Edinburg on October 2, 2017. The experiments were performed in a two-day process due to lack of time. Instructions were given by our TA on where to find the substances (guaiacol under the fume hood, turnip extract, peroxide, and distilled water were placed on our lab tables in dropper bottles, along with the spectrophotometer) and were told to get started. In activity 1 we will be testing 3 concentrations of an enzyme (0.5 ml, 1.0 ml, and 2.0 ml of turnip extract). To quantify the rate of reaction in turnips, guaiacol will be used as the color reagent. Guaiacol is oxidized when it encounters peroxide, allowing light at 470 nm to be absorbed and allowing us to measure the absorbance. In the first activity from experiment day 1, three test tubes were obtained and two clean cuvettes from our lab TA, and placed in a test tube rack on our lab tables. We used one of the test tubes to make the control, another to make the substrate and the last one to make the enzyme. We did this process 3 times to test the effects of the low enzyme concentration, medium enzyme concentration, and high enzyme concentration on the enzyme reaction rate. For the low enzyme concentration, on the control test tube we added 1.0 ml of guaiacol, 0.5 ml turnip extract, 0 ml of peroxide and 8.5 ml of distilled water, getting a total volume of 10 ml in the test tube. For the low enzyme concentration, on the

Turnips and horse radish roots are rich source of this enzyme. In this experiment, we would carry out a reaction between hydrogen peroxide and guaiacol which is colorless dye, using peroxidase as a catalyst, to produce water and an oxidized form of guaiacol which is brown. The formation of brown color would serve as an indicator that the breakdown of Hydrogen Peroxide took place. The enzyme activity would be directly proportional to the brown color intensity. The color intensity would be measured using a spectrophotometer and standardized to find the corresponding concentration for each absorbance unit.

Background and Introduction: Enzymes are proteins that process substrates, which is the chemical molecule that enzymes work on to make products. Enzyme purpose is to increase the rate of activity and speed up chemical reaction in a form of biological catalysts. The enzymes specialize in lowering the activation energy to start the process. Enzymes are very specific in their process, each substrate is designed to fit with a specific substrate and the enzyme and substrate link at the active site. The binding of a substrate to the active site of an enzyme is a very specific interaction. Active sites are clefts or grooves on the surface of an enzyme, usually composed of amino acids from different parts of the polypeptide chain that are brought together in the tertiary structure of the folded protein. Substrates initially bind to the active site by noncovalent interactions, including hydrogen bonds, ionic bonds, and hydrophobic interactions. Once a substrate is bound to the active site of an enzyme, multiple mechanisms can accelerate its conversion to the product of the reaction. But sometimes, these enzymes fail or succeed to increase the rate of action because of various factors that limit the action. These factors can be known as temperature, acidity levels (pH), enzyme and/or substrate concentration, etc. In this experiment, it will be tested how much of an effect

However, the rate of reaction only increases for a certain period of time until there is lesser substrate molecules than the enzyme molecules. The increase of enzyme concentration does not have effect if there are lesser substrate molecules than enzyme molecules initially.

The purpose of this experiment is to learn the effects of a certain enzyme (Peroxidase) concentration, to figure out the temperature and pH effects on Peroxidase activity and the effect of an inhibitor. The procedure includes using pH5, H202, Enzyme Extract, and Guaiacol and calibrating a spectrophotometer to determine the effect of enzyme concentration. As the experiment continues, the same reagents are used with the spectrophotometer to determine the temperature and pH effects on Peroxidase activity. Lastly, to determine the effect of an inhibitor on Peroxidase, an inhibitor is added to the extract. It was found that an increase in enzyme concentration also caused an increase in the reaction rate. The reaction rate of peroxidase increases at 40oC. Peroxidase performed the best under pH5 and declined as it became more basic. The inhibitor (Hydroxy-lamine) caused a decline in the reaction rate. The significance of this experiment is to find the optimal living conditions for Peroxidase. This enzyme is vital because it gets rid of hydrogen peroxide, which is toxic to living environments.

Enzymes are types of proteins that work as a substance to help speed up a chemical reaction (Madar & Windelspecht, 104). There are three factors that help enzyme activity increase in speed. The three factors that speed up the activity of enzymes are concentration, an increase in temperature, and a preferred pH environment. Whether or not the reaction continues to move forward is not up to the enzyme, instead the reaction is dependent on a reaction’s free energy. These enzymatic reactions have reactants referred to as substrates. Enzymes do much more than create substrates; enzymes actually work with the substrate in a reaction (Madar &Windelspecht, 106). For reactions in a cell it is

ABSTRACT: Enzymes are biological catalysts, and responsible for most of the chemical reactions taking place in living organisms. Enzymes speed up a chemical reaction and are not consumed during the experiment. Three experiments were performed during this lab. Experiments were performed to examine the specificity of an enzyme, effect of temperature on enzyme, and the effect of an inhibitor on an enzyme. First experiment was to demonstrate the specificity of an enzyme.

Factors include temperature, pH, inhibitors and activators-all of which will be tested and observed for in this lab. The rate of enzyme-catalyzed is affected by concentrations of both substrate and enzyme. Increasing the temperature on a reaction increases its molecular movement. The rate of an enzyme-catalyzed reaction increases with temperatures but only up until the point of optimum temperature-the highest point before the eventual decline. Below the optimum temperature, the hydrogen bonds and hydrophobic interactions that make the enzyme its given shape can no longer be supported by its flexibility.

The purpose of this experiment was to record catalase enzyme activity with different temperatures and substrate concentrations. It was hypothesized that, until all active sites were bound, as the substrate concentration increased, the reaction rate would increase. The first experiment consisted of five different substrate concentrations, 0.8%, 0.4%, 0.2%, 0.1%, and 0% H2O2. The second experiment was completed using 0.8% substrate concentration and four different temperatures of enzymes ranging from cold to boiled. It was hypothesized that as the temperature increased, the reaction rate would increase. This would occur until the enzyme was denatured. The results from the two experiments show that the more substrate concentration,

Enzyme catalysis is dependant upon factors such as concentration of enzyme and substrate, temperature and pH. These factors determine the rate of reaction, and an increase in temperature or pH above the optimum will

The purpose of this lab report is to investigate the effect of substrate concentration on enzyme activity as tested with the enzyme catalase and the substrate hydrogen peroxide at several concentrations to produce oxygen. It was assumed that an increase in hydrogen peroxide concentration would decrease the amount of time the paper circle with the enzyme catalase present on it, sowing an increase in enzyme activity. Therefore it can be hypothesised that there would be an effect on catalase activity from the increase in hydrogen peroxide concentration measured in time for the paper circle to ride to the top of the solution.

In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or

The purpose of this lab is to explore the effects of enzyme concentration, substrate concentration, temperature, and inhibitors on reaction rate, respectively. To test each of these factors, four activities were completed. The enzyme from Turnip Extract and the substrate Peroxide were tested. The turnip extract was tested at the following concentrations: .5ml, 1.0ml, and 2.0ml. Peroxide was tested at the following concentrations: 0.1ml, 0.2ml, and 0.4ml. In order to understand the effect of temperature on reaction rate the following temperatures were tested: 4C, 23C, 37C, and 60C. To achieve the desired temperatures, an ice bath and water baths were utilized. Lastly, the effect of the inhibitor Hydroxylamine was tested in the following amounts: 0 drops, 1 drop, and 5 drops. In the experiment, Guaiacol was used to determine the rate of reaction by the absorbance measured. When Guaiacol is oxidized it has a brownish color, the presence of the brownish color indicates Peroxide (substrate of peroxidase) has undergone the reaction and has been reduced to water. By this, the reaction rate can be determined by the color change or absorbance over time. For each activity, a spectrophotometer was used to measure the absorbance. After that, the data for each activity was translated to graph where linear regression was used to analyze the effects. The results of the enzyme concentrations on reaction rate are the following: low enzyme concentration (0.5 ml) had a reaction rate of