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Werdel per 2

Wes Bala

on 18 September 2012

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Transcript of Experiment

Enzyme Activity Introduction We want to find the effect enzyme concentration has on enzyme activity. Purpose 1. Acquire your materials.
2. Fill 1 cuvette with 9.7 mL of distilled water, .2 mL of Hydrogen Peroxide, and .1 mL guaiacol. This is the blank.
3. But the blank into the spectrophotometer and set the spectrophotometer to 0.
4. Take the blank out.
5. Label one test tube "Substrate", one "Mixing", and the rest, "Enzyme 1," "Enzyme 2," etc.
6. Fill the Substrate test tube with 4.7 mL distilled water, .2 mL Hydrogen Peroxide, and .1 mL guaiacol.
7. Fill the Enzyme 1 test tube with 4.0 mL distilled water, and 1.0 mL of turnip peroxidase.
8. Pour the Substrate test tube and Enzyme 1 test tube into the Mixing test tube.
9. Stretch the Parafilm over the opening of the Mixing test tube
10. Swirl around the mixture inside of the test tube to mix it.
11. Pour the mixture into a cuvette.
12. Place the cuvette into the spectrophotometer.
13. Take the initial reading on the spectrophotometer using the "absorbance" scale. Hypothesis Variables, Controls, Constants Materials Procedure Data Others
6 test tubes
5 cuvettes
1 5cc syringe
1 1cc syringe
1 peroxidase beaker
1 pack of lens paper
1 spectrophotometer
1 stop watch
5 parafilm pieces
1 test tube tray If we increase enzyme concentration, then the rate of reaction will also increase. Furthermore, if we decrease enzyme concentration, then the rate of reaction will also decrease, because the enzyme concentration affects the rate of reaction. Independent Variables: Time and Enzyme Concentration

Dependent Variables: Absorbance

Control: 4.0 mL Turnip Peroxidase

Constants : Temperature, pH,
Substrate concentration, salinity, and time. 14. Take a reading every 20 seconds for 5 minutes.
15. After 5 minutes take out the cuvette
16. Place the Blank back into the spectrophotometer and set the Spec. to 0 again.
17. Clean your Substrate test tube and Mixing test tube.
18. Fill the Substrate test tube with the same mixture as before.
19. Fill the Enzyme 2 test tube with the amounts of distilled water and turnip peroxidase in the chart from the Materials list.
20. Mix the Substrate test tube content with the Enzyme 2 test tube contents in the Mixing tube, also using the Parafilm, like before.
21. Pour the mixture into a clean cuvette.
22. Place the cuvette into the spectrophotometer.
23. Take the Initial reading.
24. Take a reading every 20 seconds for 5 min.
25. Repeat steps 15-24 for the rest of the 3 remaining enzyme concentration measurements. Conclusion Rate of Reaction per second Seconds Enzyme Concentration Enzyme number Recorded Data Jason Allison
Wes Bala
Bill Bui
Michelle Bui
Jonathan Hwang Pictures Test tubes Cuvettes Peroxidase Test tube rack Parafilm 5cc syringe beaker of H2O Guiacol 1cc syringe pipette stop watch hydrogen
peroxide There are many abiotic and biotic factors that can affect the rate of an enzyme reaction.

Some abiotic factors include: Temperature, pH, and Salinity
While some biotic factors include: Enzyme Concentration and Substrate Concentration.

Abiotic and Biotic factors can either slow down or speed up a reaction. Only at certain levels of these factors (ex. certain temp., certain substrate concentration) can an enzyme react at its maximum production speed. A. Our hypothesis proved to be correct. As we increased the enzyme concentration in each trial, the rate increased, and when we decreased the concentration, the rate also decreased. The End

Thanks for Watching! Period:2 C. A real life of example of an increase in the rate of reaction by enzyme concentration can be seen though pepsin. As more of the enzyme is present in the stomach, the faster proteins will be broken down and digested. Too little of the enzyme could cause the body to digest at a slower rate. B. Within this experiment you will find that every single time you increase the concentration of the enzyme the rate increases, but when does the rate level out? Another experiment could be done to find out the maximum efficiency of the enzyme.
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