Enyzme activity lab report PDF

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Enzymatic activity-exploration lab report Dependence of catalase activity on temperature Aim Aim of this experiment is to investigate the dependence of catalase activity on rise of the temperature. Background Catalase is an enzyme that breaks hydrogen peroxide into water and oxygen. Hydrogen peroxide is a common byproduct of metabolic reactions occurring in an environment where water and oxygen are present, but it is toxic to cells. Cells produce hydrogen peroxide (H2O2) as a toxic by-product of normal cellular reactions. The enzyme catalase quickly breaks down hydrogen peroxide into water and oxygen. In other words, catalase protects cells from the toxic effects of hydrogen peroxide. All aerobic cells produce catalase. One molecule of catalase enzyme may work on 40 million molecules of hydrogen peroxide per second.

In this experiment we are going to investigate the dependence of catalase activity on temperature. Enzyme activity increases as temperature increases, often doubling with 10°C rise. This happens because of collisions between substrate and active site. They start to happen more frequently at higher temperatures due to faster molecular motion. But at too high temperature enzymes denature and stop working. Research question How does the catalase activity depend on the temperature? Variables Table 1: Table with types of variables

Type of variable Independent variable Dependent variable

Variable Temperature(°C) Catalase activity

Table 2: Control variable table

Controls Amount of yeast extract

Method of control Always the same amount of

Why? I am going to use the same

yeast used, no additives

Calibration of instruments

Each test preformed with the same calibration

Same hydrogen peroxide percentage and amount

Always the same percentage and amount of hydrogen peroxide used

amount of yeast because if I added more or less yeast it would affect the enzyme activity and I would not get valid results Comparing the samples with the same calibration gives us precise results and ability to compare them side to side Always the same percentage and amount of hydrogen peroxide used because if we used less or more results would vary and be invalid

Hypothesis My hypothesis is that catalase activity will rise with the rise of temperature. Null hypothesis Null hypothesis is that catalase activity would not depend or be affected with rise of temperature. Materials              

Ruler (precision to 0.1cm) Thermometer (precision to 0.1°C) Bunsen burner** Hydrogen peroxide solution(30%)* Extract of yeast(5x1 cm of yeast for each test tube) Wax pencil Crushed ice 6 clean test-tubes Beaker(250 ml) Water Stand Clamp Wire gauze Tripod

Procedure 1. Put on all the protective equipment. 2. Mark test tubes with wax pencil on 1cm and 2cm, for that use the ruler( precision to 0.1cm). One test tube mark only with 1cm so you can measure, 1 cm of hydrogen peroxide solution will be later added to the yeast.

3. Place Bunsen burner on the table and put the stand next to it. 4. Above the Bunsen burner put the tripod and lay wire gauze on top of it. 5. Take the beaker(250mL) and put crushed ice in it, and then place it on the top of the wire gauze. 6. Take clean marked test tube and fill it with 1cm of yeast and put the clamp on the stand. Attach the test tube to the clamp. 7. Put thermometer in crushed ice. After that put the test tube with the yeast in beaker with crushed ice and wait 3 minutes. 8. After 3 minutes add the 1 cm of hydrogen peroxide and wait till the reaction is finished. 9. After it is finished take the ruler and precisely measure how much foam has grown and write it in the raw table data below. 10. Repeat steps 6.-9. But instead of crushed ice use, 15°C, room temperature, 50°C, 70°C water temperature. 11. After usage, wash everything that was used. Risk assessment *30-50% Hydrogen peroxide is an irritant of the eyes, mucous membranes, and skin. Use protective googles and gloves. **You could be aware that glass is going to be hot and be careful with handling Bunsen burner and hot glass, use gloves and folded paper to grab the hot glass.

Figure 1: Sketch of the apparatus

Table of raw data Temperature (±0.1°C)

Height of foam(±0.1cm)

Trial 1 Trial 2 Trial 3 Trail 4 Trial 5

Bibliography 1. “Brent Cornell.” BioNinja, ib.bioninja.com.au/standard-level/topic-2-molecularbiology/25-enzymes/enzyme-experiments.html. 2. Libretexts. “3.4: Catalase.” Biology LibreTexts, Libretexts, 7 Sept. 2020, bio.libretexts.org/Bookshelves/Ancillary_Materials/Laboratory_Experiments/Micr obiology_Labs/Microbiology_for_Allied_Health_Students:_Lab_Manual/3.04:_Cat alase.

3. Libretexts. “6.2: Catalase.” Biology LibreTexts, Libretexts, 14 July 2020, bio.libretexts.org/Bookshelves/Ancillary_Materials/Laboratory_Experiments/Gen eral_Biology_Labs/Book:_Unfolding_the_Mystery_of_Life__Biology_Lab_Manual_for_NonScience_Majors_(Genovesi_Blinderman__Natale)/6:_Enzymes/6.2:_Catalase....