Toothpickase Lab.docx

Regina FortuneRebecca JohnsonIvette RustandKhadijah Isaacsperiod 711-5-2013ToothPickase LabObservations - We are using toothpicks to simulate enzymes, substrates, and active sites. We are seeing how temperature and substrate concentration affect enzyme function. We are also measuring the rate of product formation in an enzyme-facilitated reaction.Questions - How does temperature and substrate concentration affect enzyme function? What is the rate of product formation in an enzyme-facilitated reaction?Hypothesis - Part A - As time progresses the amount of toothpicks that are broken will increase but the rate in which the toothpicks break would decrease. Part B - The higher concentration of toothpicks, the more toothpicks broken. Also, as concentration increases, the rate in which the toothpicks are broken will increase because it will be easier to find toothpicks instead of paper clips. Part C - It will take a longer amount of time to break the toothpicks after using the cold water. Independent variable - Part A:The amount of time within each interval Part B: concentration of toothpicksPart C: Use of cold waterDependent variable -Part A: The amount of toothpicks broken within each interval Part B: The amount of toothpicks broken within each interval. Part C: The amount of time it takes to break the toothpicksControls - Part A: The bowl, the amount of toothpicks (whole or broken) within the bowl Part B: The bowl, the amount of toothpicks, amount of paper clips, time spent breaking toothpicksPart C: The amount of toothpicks used.Materials Part A: 100 toothpicks per team ,bowl, clock/watch with a second hand ,PencilPart B: 1 box of toothpicks per team, 100 paper clips, clock/watch with a second hand, PencilPart C: 10 toothpicks per team, ice and ice bucket, clock/watch with a second hand, PencilProcedure Part A: 1. Count out 100 unbroken toothpicks into a bowl on your desk.2. Have one person in the group serve as the timer, have one person serve as the recorder, and have another person in your group act as the enzyme or toothpick-ase.3. The person acting as the enzyme is to break toothpicks without looking at the bowl and all of its products (broken toothpicks). All broken toothpicks must remain in the bowl along with the unbroken toothpicks, & you cannot re-break a broken toothpick!.4. The experiment is conducted in 10 second intervals.5. WITHOUT LOOKING AT THE BOWL, break as many toothpicks as you can in 10 second intervals and record this on the data table. Broken toothpicks should be kept in the bowl with unbroken toothpicks because products & reactants mix in metabolic reactions. DO NOT BREAK TOOTHPICKS ALREADY BROKEN! Remember when counting, two halves equal a whole broken toothpick! 6. Do another 10 seconds of breaking (total of 20 seconds now), and then count & record the number of toothpicks broken.7. Do another 10 seconds (thirty seconds total now) more of breaking and count and record the number of toothpicks broken.8. Continue breaking toothpicks for these total time intervals ( 60, 120, and 180 seconds). REMEMBER TO ALWAYS THROW BROKEN TOOTHPICKS BACK IN THE PILE (because products & reactants stay mixed in reactions), BUT DONT RE-BREAK THEM(the enzyme has already acted on the substrate! 6. Graph the number of toothpicks broken as a function of time (10, 20, 30, 60, 120, & 180 seconds.) Be sure to title your graph and to label the x and y-axis.

Part B: Remove the broken toothpicks from the shallow bowl. Place 100 paper clips in the empty bowl. The paper clips represent a solvent in which the toothpicks are dissolved. Different concentrations are simulated by mixing different numbers of toothpicks in with the paper clips.For the first trial, place 10 toothpicks in the bowl with the paper clip. Mix them up. The enzyme has 20 seconds to react (break as many toothpicks as possible). Remember the enzyme breaks the toothpicks without looking at the bowl and all of the products (broken toothpicks) must remain in the bowl. Remember toothpicks can only be digested once; do not break toothpicks already broken! Record the number broken at a concentration of 10.Remove the broken toothpicks and repeat with concentrations of 20, 30, 40, 50, 60, 70, 80, 90, and 100 toothpicks, each time mixing them with the 100 paper clips.Graph the results.Discuss your results and explain why the rates were different at different concentrations. Summarize the effect of substrate concentration on enzyme action.Part C:1. Select 10 toothpicks. Time how long it takes to break the 10 toothpicks as fast as you can.2. Place your hands in the pail of iced water for 10 minutes. Repeat step 1.3. Calculate the rate of enzyme action in toothpicks per second. Compare the two rates.4. Discuss your results and explain why the rates were different at different temperatures. Summarize the effect of temperature on enzyme action.DataData Table #1Total Time (seconds)Number of toothpicks broken

104

20(additional 10 sec)5

30(additional 10 sec)5

40(additional 10 sec)12

50(additional 10 sec)14

Graph #1

Data Table #2

Time (seconds)Toothpick ConcentrationNumber of toothpicks broken

20104

20206

20307

20407

20508

Graph #2

Data #3Time (seconds)# of toothpicks brokenRate (toothpicks/second)

Before Cold Water3510.29

After Cold Water6510.15

ConclusionPart A: As time increased from 10 seconds to 50 seconds, the number of toothpicks broken increased from 3 to 16. However, as the time increased from 10 seconds to 50 seconds, the rate went from breaking .4 toothpicks per second to breaking .28 toothpicks per second. This data supports the hypothesis that as time increases, the number of toothpicks broken will increase but the rate at which they are broken will decrease. This was mainly due to the fact that as time increased, more toothpicks were broken and it took longer to sort through the broken ones to find an unbroken toothpick. This part of the lab demonstrates the rate at which an enzyme functions. As time passes, the amount of substrates an enzyme binds to increases but the rate at which it works decreases. Part B: As the concentration of toothpicks increased from 10 toothpicks to 50 toothpicks, the amount of toothpicks broken increased from 4 to 8. This data supports our hypothesis that as concentration increases, the amount of toothpicks broken will increase and the rate at which they are broken will increase. This increased rate is seen when .2 toothpicks were broken per second at 10 toothpick concentration and .4 toothpicks were broken per second at a 50 toothpick concentration. This data suggests that as substrate concentration is increased, the rate at which enzymes function is also increased because it is easier to find substrates to bind with. Part C: In this experiment, it took 35 seconds to break 10 toothpicks before putting the enzymes, or the breakers hands, in cold water. After putting the breakers hands in cold water, they were able to break 10 toothpicks in 65 seconds. This supports our hypothesis that it will take longer to break the toothpicks after using the cold water. This suggests that enzymes function at a slower rate when the temperature is drastically lowered. Questions1.What happens to the reaction rate as the supply of toothpicks runs out? The reaction rate decreases, because the amount of broken toothpick increases while the amount of unbroken toothpicks decreases. This makes it harder to find the unbroken toothpicks because they are surrounded by all of the unbroken ones. 2. What would happen to the reaction rate if the toothpicks were spread out so that the "breaker" has to reach for them?The reaction rate would decrease because it would be much more difficult to find and break the toothpicks. It would also be much more slower since the time adds on and we have to search for them.

3. What would happen to the reaction rate if more toothpicks (substrate) were added? The reaction rate would increase because there are more toothpicks to break ,it would also help because there is a slight chance we won't take a broken one instead there are more unbroken ones.4. What would happen to the reaction rate if there were two "breakers" (more enzymes)?The reaction rate would be much more higher because we would be doing twice as much work as for one person breaking. 5. What happens if the breaker wears bulky gloves (active site affected) when picking up toothpicks?It would be much harder to find and break and due to the size being so big and therefore affecting the time it takes (concentration) and how many we brake. 6. Explain what would happen to an enzyme-facilitated reaction if temperature were increased. Be sure to include the effect if temperature were increased to 100C. Increasing temperature of an enzyme-facilitated reaction would slightly speed up the reaction because molecules would move around more quickly. But notice that word "slightly". Higher temperatures denature enzymes. If the temperature were increased to 100 C, the protein (enzyme) would have been denatured or destroyed long before it got that hot. 7. What is the optimal temperature (C) for enzymes functioning in the human body?Optimal temperature for enzyme action in the human body should be normal body temperature - 37 C.enzymes usually function best at body temperature (that's what they're designed for) or 37 degrees C. The work faster at higher temperatures, but if they are heated up to much, the proteins within the enzymes will denature, causing them to be biologically inactive and useless.