enzyme lab
DESCRIPTION
TRANSCRIPT
Enzyme Student Training
By Quanina Quan UCSD ScienceBridge
Objective
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.
The Enzyme Tyrosinase
Tyrosinase is an enzyme that causes pigmentation in apples, skin, hair, etc.
The Enzyme Tyrosinase
The Enzyme Tyrosinase
Albinism
The Reaction
General Enzyme-Substrate Reaction
Enzyme + Substrate = Product + Enzyme
General Tyrosinase ReactionTyrosinase+ L-DOPA = Dopachrome + Tyrosinase
+ =
The Lab
3 Main Parts of the Tyrosinase Enzyme Lab
1. Enzyme Extraction2. Control Reaction3. Experimental Reaction
The Lab – Enzyme Extraction
1a. Break open the cell to free Tyrosinase
The Lab – Enzyme Extraction
1b. Filter out the solid chunks of mushroom
The Lab – Control Reaction4 roles:- Mixer - Reader- Timer - Recorder
Tyrosinase+ L-DOPA = Dopachrome + Tyrosinase
+=
0.1 mlENZ CTL CTL
The Lab – Experimental Trials
3 Different Experimental Trials
- pH- Inhibitor- Temperature
The Lab – Experimental TrialpH
Tyrosinase+ L-DOPA = Dopachrome + Tyrosinase
+=
0.1 ml
- Test the pH of L-DOPA- Test the pH of L-DOPA after you add HCl- Start the reaction
HCl
?ENZ EXP EXP
The Lab – Experimental Trial
Inhibitor- Add Inhibitor- Start the reaction
Tyrosinase+ L-DOPA = Dopachrome + Tyrosinase
+=
0.1 ml
Inhibitor
?EXP EXPENZ
The Lab – Experimental TrialTemperature
-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
L-DOPA
=
?
EXP EXP
EXP
Graphing!
X = time in secondsY = mM of Dopachrome
10 30 50 80 120
160
240
360
0
0.5
1
1.5
2
2.5
Sample Control Enzyme Reaction
Control Enzyme Reaction
Why Study Enzyme Reaction Rates?
Myasthenia Gravis Dog Video Cone sea snails Video
(toxin inhibiting receptors)