Turnip Peroxidase Lab Report

Most enzymes work best at body temperature, higher temps will cause the enzyme to no longer work properly

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 preparation for the experiment started by gathering the solutions of enzyme Peroxidase, substrate hydrogen peroxide, the indicator guaiacol and distilled water. Two small spectrometer tubes and three large test tubes with numbered labels. In addition, one test tube rack, one pipet pump and a box of kimwipes were also gathered. Before the experiment, the spectrometer must be set up to use by flipping the power switch to on. Following, the machine was warmed up for 10 minutes and the filter lever was moved to the left. In addition, I set the wavelength to 500 nm with the wavelength control knob. Before the experiment, I had to create the blank solution by pipetting 0.1 ml of guaiacol, 1.0 ml of turnip extract and 8.9 ml water into tube #1. Following the creation of the blank, a control 2% solution was created.

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

One of the best-studied peroxidases is horseradish peroxidase (HRP), which has a heme-iron co-factor. In most heme-peroxidases the iron atom in the active center undergoes a reversible change of its oxidation state. The reaction proceeds in three distinct steps. In first step, the resting state high-spin Fe(III) is present, which is oxidized by hydrogen peroxide to form an unstable intermediate called compound I (Co-I) with Fe(IV), releasing water in the process. Compound I is not a classical enzyme–substrate complex, but rather a reactive intermediate with a higher formal oxidation state (­5 compared with ­3 for the resting enzyme). Thus, compound I is capable of oxidizing a range of reducing substrates. This reactive intermediate oxidizes

3. We measured 2.5 mL of turnip peroxidase (the enzyme) and 10 mL of neutral buffer (pH 7) with a syringe and disposed it into test tube ENB.

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.

Students will be observing normal catalase reaction, the effect of temperature on enzyme activity, and the effect of pH on enzyme activity in this experiment. The enzymes will all around perform better when exposed in room temperature than when it is exposed to hot and cold temperatures. This is based on the fact that the higher the temperature, the better the enzymes will perform, but as the temperature reaches a certain high degree, the enzymes will start to denature, or lose their function.

Enzymes are catalysts that lower the activation energy required to start a biological reaction and affects the rate of the rection. The enzyme can become denatured when its surroundings change in temperature or pH. Molecules in the environment could also affect the enzymatic activity. Inorganic substances known as cofactors and organic molecules known as coenzymes can enhance or inhibit the enzymes activity. The cofactors or coenzymes can act to activate or inhibit the enzymatic activity. Peroxidase is a catalyst that catalyze substrate oxidation when a peroxide is present. The indicator guaiacol is easily oxidized by peroxidases. Guaiacol is used to indicate if the peroxidase enzyme is present in a solution. Turnip peroxidase breaks down hydrogen peroxide into water and

The purpose of this study is to determine the effects of temperature variance in the rate of the enzymatic activity of the enzyme peroxidase, which converts hydrogen peroxide into water (Moore and Vodopich, 77). Materials and

As the temperature increases, so will the rate of enzyme reaction. However, as the temperature exceeds the optimum the rate of reaction will decrease.

Hypothesis: I believe the rate of reaction will speed up as the temperature increases until it reaches about 37oC, which is the body temperature, where it will begin to slow down and stop reacting. I believe this will occur because enzymes have a temperature range at which they work best in and once the temperature goes out of this range the enzyme will stop working.

By observing the constructed graph, it is shown that the average rate of change initially increased as the temperature increased, but when temperatures increased past 50°C, the average rate of the reaction began to decrease until the 60°C point. therefore, the height of the reaction initially increased until the optimal point which was around 50°C and further decreased as the temperature was increased. This graph is very similar to what was predicted as the hypothesis. These results outline the effect of temperature on enzyme activity, and it is obvious that changes in temperature do have an effect on the enzyme catalase.

Enzymes are macromolecules that act as a catalyst, and it’s a chemical agent that accelerates the reaction without being consumed by the feedback or the results (Campbell and Reece, 2005). After the adjustment by the enzymes, the chemical movement through the pathways of metabolism will become awfully crowded because many chemical reactions are taking a long time (Campbell and Reece, 2005). There are two kinds of reactions in nature. The first one is Catabolic reaction and the second one is Anabolic reaction. Catabolic reactions are large molecules that are broken up into smaller molecules (Ahmed, 2013). Anabolic reactions are small molecules that join to make larger molecules, like polymerization (Ahmed, 2013). If you