The document summarizes an experiment to test the reaction rate of the enzyme tyrosinase under different conditions. Tyrosinase is extracted from portobello mushrooms and its reaction with L-DOPA is tested under normal conditions and compared to when exposed to inhibitors, increased temperature, or changes in pH. The experiment involves extracting the enzyme, performing a control reaction, and then experimental reactions under different pH levels, inhibitors, or temperatures to see how these factors affect the reaction rate of tyrosinase.

1. By Quanina Quan UCSD ScienceBridge

2. To extract the enzyme Tyrosinase from a
Portobello mushroom in order to test the
reaction rate of the enzyme in normal
conditions and compare it to conditions where
the enzyme is exposed to inhibitors, increased
temperature, or changes in pH.

3. Tyrosinase is an enzyme that causes
pigmentation in apples, skin, hair, etc.

5. Albinism

6. General Enzyme-Substrate Reaction
Enzyme + Substrate = Product + Enzyme
General Tyrosinase Reaction
Tyrosinase + L-DOPA = Dopachrome + Tyrosinase
=
+

7. 3 Main Parts of the Tyrosinase Enzyme Lab
1. Enzyme Extraction
2. Control Reaction
3. Experimental Reaction

8. 1a. Break open the cell to free Tyrosinase

9. 1b. Filter out the solid chunks of mushroom

10. 4 roles:
- Mixer - Reader
- Timer - Recorder
Tyrosinase + L-DOPA = Dopachrome + Tyrosinase
0.1 ml
ENZ CTL CTL
=
+

11. 3 Different Experimental Trials
- pH
- Inhibitor
- Temperature

12. pH
- Test the pH of L-DOPA
- Test the pH of L-DOPA after you add HCl
- Start the reaction
Tyrosinase + L-DOPA = Dopachrome + Tyrosinase
0.1 ml
ENZ EXP EXP
+
= ?
HCl

13. Inhibitor
- Add Inhibitor
- Start the reaction
Tyrosinase + L-DOPA = Dopachrome + Tyrosinase
0.1 ml
EXP EXP
ENZ
+
= ?
Inhibitor

14. Temperature
-Heat 0.1 ml of L-DOPA in EXP tube for 1 min. in
water
- Add L-DOPA and start the experiment
0.1 ml Tyrosinase ?
EXP EXP
=
EXP
L-DOPA

15. X = time in seconds
Y = mM of Dopachrome
Sample
2.5 Control Enzyme Reaction
2
1.5
1 Control Enzyme
Reaction
0.5
0
10 30 50 80 120 160 240 360

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