Catalase in potato and hydrogen peroxide concentration

Introduction Hydrogen peroxide (H2O2) is a by-product of respiration and is made in all living cells. Hydrogen peroxide is harmful and must be removed as soon as it is produced in the cell. Cells make the enzyme catalase to remove hydrogen peroxide. This investigation looks at the rate of oxygen production by the catalasein pureed potato as the concentration of hydrogen peroxide varies.

The reaction of catalase in the decomposition of hydrogen peroxide is: 2 H2O2 2 H2O + O2

Catalase is used in the food industry for removing hydrogen peroxide from milk prior to cheese production. Another use is in food wrappers where it prevents food from oxidizing. Catalase is also used in the textile industry, removing hydrogen peroxide from fabrics to make sure the material is peroxide-free.

A minor use is in contact lens hygiene - a few lens - cleaning products disinfect the lens using a hydrogen peroxide solution; a solution containing catalase is then used to decompose the hydrogen peroxide before the lens is used again. Recently, catalase has also begun to be used in the aesthetics industry. Several mask treatments combine the enzyme with hydrogen peroxide on the face with the intent of increasing cellular oxygenation in the upper layers of the epidermis.

Aim: To investigate the effect of Catalase on H2O2 and the volume of O2 produced.

Hypothesis: As the concentration of Catalase increases from 2v0.25 cm to 2v2 cm potato cylinders, the volume of O2 will also increase approximately twice.

Variables: a) Dependant: Volume of O2 b) Independent: Temperature of environment c) Controlled: volume of H2O2, mass or volume of disk potatoes, concentration of H2O2, volume of water in the collector.

Apparatus & Chemicals: y y y Hydrogen peroxide, 20 vol Potato Cylinders, fresh, 2v0.25 cm, 2v0.50 cm, 2v1.00 cm, 2v2.00 cm Rubber bung, 1-holed, to fit test tube delivery tube in the hole (connected to 50 cm rubber tubing)

y y y y y

Access t suit le si 4 Test tubes 2 Syri e (5 cm3) 1 Measuri

of water

cyli er, 5 cm3

2 Stopclock/ stopwatch

Heal

& Safety

Wear eye protection and cover clothing when handling hydrogen peroxide. Wash splashes of pureed potato or peroxide off the skin immediately. Be aware of pressure building up if reaction vessels become blocked. Take care inserting the bung in the conical flask it needs to be a tight fit, so push and twist the bung in with care. SAFETY: Wear eye protection and protect clothing from hydrogen peroxide. Rinse splashes of peroxide and pureed potato off the skin as quickly as possible. The tube will get hot during the reaction. So be aware.

Method:

We put the potato cylinders inside the test tubes marked showing the length of the cylinders putted inside them.

Then we prepare the measuring column as shown below.

Before putting the bung on the tube, we insert 5 cm 3 of H2 O2 in the tube.

At the same time that we input the H2 O2 we start measuring the time with stop clock.

y y y y y

We close the tube with the bung immediately as we pour the H2 O2. We take time for 5 minutes. Then we measure the volume of oxygen collected and right it down. Results were recorded onto the table, where it was analysed to draw a graph and make a conclusion. The desk and equipment were cleaned and put where they belong. Results:

Si e of a potato disc (cm) 2*0.25 2*0.50 2*1.00 2*2.00

Oxygen collected per 10 min with 5cm3 of hydrogen peroxide 6.5 6.0 6.5 7.0 7.5 7.0 10.0 8.5 9.0 9.0 11.0 10.0

Mean 6.3 7.2 9.2 10.0

Evaluation: As our group was working individually, we noticed that a human factor has influenced our final results, as a one person was responsible for tube cap removal and the other one had to measure time. At first this minor problem does not seem like it could affect our results significantly, but it lead our group to major issue which due to all this kept collected oxygen bubbles inside the tube and didnt let us to collect oxygen instantly as hydrogen peroxide reacted with catalase. However this issue can be removed by better equipment. I think that the number of repeat results taken shows that the data obtained was reliable, since three repeat experiments is enough to prove the basic idea behind a hypothesis. However, three is a relatively low number of experiments, and so I do not think that numerical conclusions can be calculated from the data for example, it would be unwise to base a formula to find the rate of reaction given a particular concentration on a data set of this si e. One particular weakness of this method of conducting the investigation is that the end point is difficult to determine and open to personal opinion. A better method of conducting the experiment would be use a variable that could be scientifically measured. Another problem is that the otato also includes other enzymes and also solid parts; therefore it was better to turn the potato into mash potato and then concentrate it into liquid form to get a higher rate and lower error. Also with better equipments it would be better to extract pure catalase from the potatos concentrate. Overall, however, the experiment was successful enough to fulfil the aim, since the effect of enzyme concentration of the hydrolysis of gelatin by protease enzymes has been investigated, and reliable conclusions have been drawn. Other investigations could be planned in future to extend the investigation, including looking at the effect

of concentration on individual types of protease enzyme. Therefore, the experiment was appropriate to its purpose and successful. Discussion: The results of our group during this experiment does not entirely agree with our hypothesis that as the volume of potato discs increases in the following sequence of 0.25, .50,1.0 and 2.0cm, the volume of oxygen produced at each stage will double. nalysis: As the size of the potato discs increased, the rising amount of enzyme catalase has produced more and more oxygen as it greater surface area of a potato was reacting with the hydrogen peroxide. Conclusion: according to the results our hypothesis was not right and could be corrected, because even though there has been an increase of collected oxygen from 6.3 to 10cm3 as the surface area increased from 0.5 to 4.0cm, it has not doubled at each stage as it was thought in the beginning. Reference:

http://www-saps.plantsci.cam.ac.uk/prac_enzymes.htm