This lab involves using a Biofuel Enzyme Kit to create our own biofuels. Through the use of certian enzymes, we will be able to create an alternate fuel source that is more efficient and better for the enviornment than burning petroleum products. Ezymes are generally proteins and what they do is speed up the reaction rate. Though enzymes are very effective, changes in salinity and pH of the substance can decrease the enzyme's effectiveness greatly. In this lab, we are trying to extract the cellulose from plant cells and harvest that for energy. In order to do this, we need to use the enzyme, cellobiase.
Procedure
Reaction Rates with/without Enzymes
1. Find your 15ml conical tube of Stop Solution and label it that
2. Find cuvettes and label 5 of them E1-E5
3. Put 500 microliters of stop solution into each cuvette
4. Label 2 empty conical tubes, Control and Enzyme Reaction
5. Pipet 2ml of 1.5 mM substrate into 15 ml conical tube labeled Enzyme Reaction
6. Pipet 1ml of 1.5mM Substrate into Control conical tube
7. Pipet 500 microliters of buffer into Control conical tube and gently mix.
8. After stirring remove 500 microliters of solution and add it to cuvette labeled start
9. Pipet 1mL of enzyme into 15mL conical tube, gently mix then START TIMER
10. At times indicated extract 500 microliters from solution and add it to correct cuvette containing the stop solution
E1=1min
E2=2min
E3=4min
E4=6min
E5=8min
11. After enzyme samples collected, remove 500 microliters of solution from Control and add it to cuvette labeled End
12. Analyze samples and clean supplies, Hey you've just created a Biofuel!
Mushroom extracts Increase Reaction Rate??
1. Weigh out approx. 1 gram of mushroom
2. Add mushroom to mortar and pestel, and for every gram of mushroom, 2 mL of extraction buffer will need to be added.
3. Using pestel, grind mushroom to "slurry"
4. Add at least 250 microliters of Mushroom extract into a 1.5 mL microcentrifugal tube and place tube in centrifuge for 2 minutes
5. Label cuvettes 1-6.
6. Pipet 500 microliters of stop solution into each cuvette
7. Label 15mL conical tube with type of mushroom and then pipette 3 mL of substrate into the tube
8. Pipette 250 microliters of enzyme extract into 15 mL conical tube of Substrate. START TIMER
9. At times remove 500 microliters of mushroon/enzyme into cuvette with stop solution.
1=1min
2=2min
3=4min
4=6min
5=8min
10. Analyze result, and clean materials that can be reused. Your done!
8. After stirring remove 500 microliters of solution and add it to cuvette labeled start
9. Pipet 1mL of enzyme into 15mL conical tube, gently mix then START TIMER
10. At times indicated extract 500 microliters from solution and add it to correct cuvette containing the stop solution
E1=1min
E2=2min
E3=4min
E4=6min
E5=8min
11. After enzyme samples collected, remove 500 microliters of solution from Control and add it to cuvette labeled End
12. Analyze samples and clean supplies, Hey you've just created a Biofuel!
Mushroom extracts Increase Reaction Rate??
1. Weigh out approx. 1 gram of mushroom
2. Add mushroom to mortar and pestel, and for every gram of mushroom, 2 mL of extraction buffer will need to be added.
3. Using pestel, grind mushroom to "slurry"
4. Add at least 250 microliters of Mushroom extract into a 1.5 mL microcentrifugal tube and place tube in centrifuge for 2 minutes
5. Label cuvettes 1-6.
6. Pipet 500 microliters of stop solution into each cuvette
7. Label 15mL conical tube with type of mushroom and then pipette 3 mL of substrate into the tube
8. Pipette 250 microliters of enzyme extract into 15 mL conical tube of Substrate. START TIMER
9. At times remove 500 microliters of mushroon/enzyme into cuvette with stop solution.
1=1min
2=2min
3=4min
4=6min
5=8min
10. Analyze result, and clean materials that can be reused. Your done!
Results/Discussion
From my observations, the solution in the cuvettes got more yellow as the time got longer. This is due to more time the enzyme has to react with cellulose reaction. Other than that, I thought that this lab was somewhat fun, but almost pointless because we didn't get to do anything with the finished product. But anyway, it was interesting to see how enzymes can break through plant cells to collect that vital energy: glucose.
Great work! A
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