Factors affecting the rate of catalase activity The extent at which environmental factors affect the rate of catalase activity was discovered in this lab. The assay system, in which a filter paper disc was dipped into the enzyme and submerged using a stirring rod in a test tube filled with 20mL of hydrogen peroxide, was used to test several enzyme factors. As the saturation of hydrogen peroxide increased the rate of reaction increased as well. When the enzyme concentration increased the rate of catalase activity increased too. When catalase was subjected to an increase of temperature changes, the rate of reaction increased. Once the protein denatured around 100ºC the catalase activity decreased. Catalase operated with a high efficiency …show more content…
Add 20ml of 3% H2O2 to a test tube 2. dilute catalase 50u/mol: 2.5 ml 100u/mol catalase and 2.5 ml of H20 3. use tweezers to dip paper disc into catalase 4. place the paper disc onto the stirring rod and submerge to the bottom of the test tube 1. Add 20ml of 3% H2O2 to a test tube and measure depth 2. dilute the catalase 20u/mol: 1ml 100u/mol catalase and 4 ml of H20 3. use tweezers to dip paper disc into catalase 4. place the paper disc onto the stirring rod and submerge to the bottom of the test tube 1. Add 20ml of 3% H2O2 to a test tube and measure depth 2. use tweezers to dip paper disc into H20 3. place the paper disc onto the stirring rod and submerge to the bottom of the test tube Part C: Temperature (assay technique) 1. create a water bath of 0ºC, a room temperature bath 23ºC, 37ºC, and boiling water bath 100ºC 2. place 5ml of catalase at 100u/mol into 4 test tubes and set one in each water bath 3. allow test tube to sit for 5 min in the respective water baths 4. add 20ml of 3% hydrogen peroxide to 4 test tubes 5. used tweezers to dip 4 pieces of filtered paper discs into separate catalase baths 6. place disc on the bottom of the stirring rod and submerge each filter paper into a hydrogen peroxide test tube Part D: pH (assay technique) 1. add 3% 20ml hydrogen peroxide to 3 separate test tubes 2. diluted catalase using 0.1 HCL and 0.1 mol NaOH to pH 1, 7, and 11.5 3.
Fill a test tube about 1/3 full with cold tap water for use in step 34.
Lab six requires students to observe the effects of pH and enzyme concentration on catecholase activity. Enzymes are organic catalysts that can affect the rate of a chemical reaction depending on the pH level and the concentration of the enzyme. As pH comes closer to a neutral pH the enzyme is at its greatest effectiveness. Also at the absorbance of a slope of 0.0122 the enzyme is affected greatly. The pH effect on enzymes can be tested by trying each pH level with a pH buffer of the same pH as labeled as the test tube and 1mL of potato juice, water, and catechol. This is all mixed together and put in the spectrophotometer to test how much is being absorbed at 420nm. As the effect on enzyme concentration can be tested almost the same way. This part of the exercise uses different amounts of pH 7-phosphate buffer and potato juice, and 1mL of catechol mixed together in a test tube. Each substance is put in the spectrophotometer at a wavelength set tot 420nm. The results are put down for every minute up to six minutes to see how enzyme concentration affects reaction rate. The results show that the pH 8 (0.494) affects the enzyme more than a pH of 4 (0.249), 6 (0.371), 7 (0.456), and 10 (0.126). Also the absorbance is greatest at a slope of 0.0122 with test tube C that has more effect on the reaction rate, than test tube A, B, and D.
Submerge the graduated cylinder in the plastic tub so that it is completely filled with water. Hold the open end of the graduated cylinder and move it vertically upside-down where the open end of the graduated cylinder is still submerged in the plastic tub. Clamp the graduated cylinder the ring stand of the lab table to keep it in place. perforate a hole in the top of the rubber cork for the solution container. Cut a straw the length of about four inches. place the straw inside of the rubber cork hole. Set up your timer for two minutes.
The hypothesis is that catalase activity will increase exponentially with higher concentrations of hydrogen peroxide until all catalase active sites are filled, in which case the
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.
1 ml of water should be added to the first test tube and make a note. In the second test tube, 1 ml of methyl alcohol should be added. In the third test tube, 1 ml of hexane must be added. Lastly, the fourth test tube will be a control.
lab bench. Place a beaker from the drawer on the stir plate. Drag the bottle of NH3 to the 5 mL graduated cylinder (the smallest one) by the sink and fill the cylinder by dropping the bottle on the cylinder. Now drag the 5 mL graduated cylinder to the beaker on the stir plate and add the 5 mL of
The purpose of this investigation is to discover the effect of pH on the activity of catalase, an enzyme which plays the integral role of converting hydrogen peroxide into water and oxygen, and discover which pH level it will work at the most efficient rate (the optimum). The original hypothesis states that that the optimum would be at a pH is 7, due to the liver, where catalase usually resides, being neutral. The experiment consists of introducing the catalase to hydrogen peroxide, after exposure to certain solutions; hydrogen peroxide, water and hydrochloric acids, all containing the adjusted pH, and measuring the height of froth formed, an observable representation of the activity of the enzyme. The final data indicated that
An Investigation on the rate of reaction of the enzyme Catalase on the substrate Hydrogen peroxide.
The Effects of Varied Temperatures, pH Values, Enzyme Concentrations, and Substrate Concentrations on the Enzymatic Activity of Catecholase
The first experiment begun by filling a 600-ml beaker, almost to the top, with water. Next, a 10-ml graduated cylinder was filled to the top with water. Once water was added to the beaker and graduated cylinder, a thumb was placed over the top of the graduated cylinder. This would ensure that no water was let out and no bubbles were let into the graduated cylinder. Next, it was turned upside down and fully submerged into the beaker. Then, a U-shaped glass tube was attained. The short end of the glass tube was placed into the beaker with the tip inside of the graduated cylinder. Next, a 50-ml Erlenmeyer flask was received. After, 10-ml of substrate concentration and 10-ml of catalase/buffer solution were placed into the flask. A rubber stopper was then placed on the opening of the flask. After adding these, the flask was held at the neck and spun softly
5. Wet the paper with distilled water to hold it in place in the funnel. Transfer all the solution and the precipitate from the beaker using a rubber policeman. Wash the precipitate with two or three 5-mL portions of distilled water. Do this by adding each portion to the beaker in which you did the precipitation to transfer any remaining
4.Measure 35mL of warm water and add them into each of the 4 test tubes at about roughly the same time. It is essential that the water is warm. Do not seal the test tube.
The independent variable in this investigation is pH. Each individual enzyme has it’s own pH characteristic. This is because the hydrogen and ionic bonds between –NH2 and –COOH groups of the polypeptides that make up the enzyme, fix the exact arrangement of the active site of an enzyme. It is crucial to be aware of how even small changes in the
Step 1 and 2 was repeated by using distilled water by replacing the test solution.