Turnip Peroxidase Lab Report

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 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.

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

In the experiment we used Turnip, Hydrogen Peroxide, Distilled Water, and Guaiacol as my substances. On the first activity, Effect of Enzyme concentration of Reaction Rate for low enzyme concentration, we tested three concentrations of the turnip extract, and hydrogen peroxide. For the Turnip Extract I used 0.5 ml, 1.0 ml, and 2.0 ml. For hydrogen peroxide we used 0.1 ml, 0.2 ml, and 0.4 ml. We used a control to see the standard, and used a control for each enzyme concentration used. The control contains turnip extract and the color reagent, Guaiacol. We prepared my substrate tubes separately from the enzyme tubes. My substrate tube

Enzyme can be defined as a protein molecule that is a biological catalyst. (Ophardt, 2003) Catalyst increases the speed of a reaction but does not have to use anything to help increase the speed. An enzyme can be determined by their properties. Enzyme are a substrate specific, substrate connects to an enzyme at the location of an active site. Enzyme is not used in a reaction and enzyme function in a good condition at the optimum temperatures and pH. (Ahmez2005) Peroxidase is a type of enzyme which is used in the experiment.

The data in proves that our hypothesis was correct. When we increased the temperature to 35°C, the the enzyme activity increased because kinetic energy increased, increasing the collisions between the substrate and the enzyme, and thus creating a higher chance of reaction. When we increased the temperature to 45°C, the enzyme activity decreased as the enzyme became denatured,because the atoms in the enzyme had enough energy to overcome the hydrogen bonds between the R groups that give the enzyme its shape From our data, we could conclude that the optimal temperature of turnip peroxidase is around 35°C and around 45°C, it will start to denature.

Peroxidase is a turnip enzyme; it is used in the oxidation of hydrogen peroxide to lower activation energy, speeding up the reaction. The activity of peroxidase is highly dependent on its environment and most importantly the pH level. Peroxidase has been the focus of many recent studies and is believed to possibly reduce swelling among other things. We conducted an experiment testing the effect different levels of pH had on the reaction rate of peroxidase. In the experiment we created different solutions all containing hydrogen peroxide, peroxidase, and guaiacol. However each cuvette contained a different pH level, 2,5,7,or

The time in the water bath was also controlled to ensure that the enzymes were left to react for the same amount of time, making the experiment

The effectiveness of peroxidase was measured in a varying pH environment. The environment’s pH can range from   1-14, 7 being neutral, 7-1 being more and more acidic towards 1, and 8-14 being more and more basic towards 14 (Raven, 2011).

After the substrate solution was added, five drops of the enzyme were quickly placed in tubes 3, 4 and 5. There were no drops of enzyme added in tubes 1 and 2 and in tube 6 ten drops were added. Once the enzyme solution has been added the tubes were then left to incubate for ten minutes and after five drops of DNSA solution were added to tubes 1 to 6. The tubes were then placed in a hot block at 80-90oC for five minutes. They were then taken out after the five minute period and using a 5 ml pipette, 5 ml of distilled water were added to the 6 tubes and mixed by inversion. Once everything was complete the 6 tubes were then taken to the Milton Roy Company Spectronic 21 and the absorbance of each tube was tested.

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.

The purpose of this report is to find out the effect of change in the Temperature, PH, boiling, concentration in peroxidase activity. Peroxidase is an enzyme that converts toxic hydrogen peroxide (H2O2) into water and another harmless compound. In this experiment we use, turnips and horseradish roots which are rich in the peroxidase to study the activity of this enzyme. The activity of peroxidase with change in temperature was highest at 320 Celsius and lowest at 40C. The activity of peroxidase was highest at a pH of 7, while it was lowest at pH of 9.Peroxidase activity was very low and constant with boiled extract, while the activity was moderate

Based on the data, the absorbance when the pH was seven was the highest. It was the lowest in an acidic environment at a pH of three, but slightly higher than the acidic in the basic environment at a pH of eleven. The rate of the reaction could be measured through the absorbance. When hydrogen peroxide breaks down, it produces oxygen gas which can react with the guaiacol to form tetraguaiacol. The solution turns brown and the darker it is, the more oxygen is produced and the greater the absorbance. At the pH of seven, the solution was the darkest, meaning the reaction proceeded quickly and the rate was higher. The reason that peroxidase functioned the best at around the pH of seven is because that is the optimal pH in cells for the enzyme. Enzymes work best at their optimal conditions. They are sensitive to their environment and tiny changes such as changes in pH can cause them to stop functioning. The shape of the enzyme or the active site can be changed so it will not attach to the substrate and become inactive. One

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

This experiment was based on the type of biomolecules known as enzymes. These molecules found in all living things, and are crucial for the removal and decomposition of harmful chemicals, the digestion of molecules in metabolism, and the synthesis of proteins, DNA, RNA, and many other organic molecules. Several key features about enzymes (which are similar to catalysts found in chemistry, only enzymes are organic molecules that are made of proteins or RNA) is their reusability; an enzyme will never be consumed in a reaction, rather it orients the molecules that would ordinarily perform the reaction correctly to minimize the energy needed and helps induce the reaction to take place faster. Most enzymes can catalyze thousands to millions of the same reactions in a second; they are very efficient at the role they perform, Also critical is the substrate, or the molecule that has to undergo the reaction, which will fit into the active site of the enzyme (a very specific geometric interaction) in order for the reaction to occur. Each enzyme is only designed to catalyze one type of reaction between very few substrate molecules; this specialization is what makes the enzymes so efficient. Of course, still being tied to a chemical reaction, the rate of the catalyzed reaction will speed up or slow down depending on several factors; these include temperature, pH, salinity, solvent, and in the case of this experiment, substrate concentration and enzyme concentration.