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ENYZYME LAB Effects of Temperature and pH on Enzyme Function Anatomy & Physiology Instructor Terry Wiseth Northland College Click To Go To Next Slide.

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Presentation on theme: "ENYZYME LAB Effects of Temperature and pH on Enzyme Function Anatomy & Physiology Instructor Terry Wiseth Northland College Click To Go To Next Slide."— Presentation transcript:

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2 ENYZYME LAB Effects of Temperature and pH on Enzyme Function Anatomy & Physiology Instructor Terry Wiseth Northland College Click To Go To Next Slide

3 ENZYME LAB Directions in performing the lab In the performance of this virtual lab understanding how the buttons will work for you should be helpful. Navigating the lab will be performed by clicking on the various buttons available in a menu you will find in the lower right hand corner. Click on the YELLOW button below to go to the next slide and learn how to navigate. Click To Go To Next Slide Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

4 ENZYME LAB Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click on each of the buttons that you see below to view description of how each button works Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

5 ENZYME LAB Clicking this button will allow you to proceed to the next slide. You will use this button often. Click on each of the buttons that you see below to view description of how each button works Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

6 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will allow you to go to the previous slide in the lab. Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

7 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will allow you to go to the slide that you have just came from. Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

8 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will allow you to quit the lab and go back to your desktop. Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

9 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will bring you to the lab index. From the lab index you can link to either Lab #1 or Lab #2. Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

10 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will bring you to Lab #1 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

11 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will bring you to Lab #2 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

12 ENZYME LAB Click on each of the buttons that you see below to view description of how each button works Clicking this button will bring you to the Enzyme Lab Tutorial. The lab tutorial provides more information about enzymes. Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click Here If You Understand How To Use All Of These Buttons Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

13 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Click To Go To Last Slide Viewed This button is utilized to return to the slide that you linked from initially. This would be analogous to using a book mark in your lab book Click To Go To Lab Report This button is utilized to link to your lab report. Using this button will bring you to pages that should be printed out so that you can include them in your lab report Click To Go To Next Slide This button is utilized to go to the next slide of the lab. This would be analogous to turning the page of your lab book You probably want to utilize this button next

14 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB This button is utilized to give information on when or how to use a specific piece of lab apparatus These buttons are utilized to go to examples of graphs or tables which may be more helpful if you print them off Turn a Machine On Click to View Data Table Click to View Graph Click To Go To Conversion Chart This button is utilized to link to conversion charts which are helpful in entering the correct values in the data tables You probably want to utilize this button next

15 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB You should print off the following pages located from these links Temperature Data Table Density Conversion Chart Graph of Temperature vs mM product Graph of pH vs mM product pH Data Table Temperature Hypothesis pH Hypothesis Temperature Conclusion pH Conclusion pH Lab Questions Temperature Lab Questions

16 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB This ENZYME LAB is divided into two Experiments Experiment #1 Experiment #1 will investigate the influence temperature has on enzyme activity Experiment #2 Experiment #2 will investigate the influence pH has on enzyme activity

17 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB INDEX Click To Go To Experiment #1 Effect of Temperature on Enzyme Activity Click To Go To Experiment #2 Effect of pH on Enzyme Activity Click To Go To The Enzyme Tutorial (Learn More About Enzymes) Go Back to A&P 2 Lab Book

18 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB #1 Temperature Influences On Enzyme Activity

19 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB #1 INDEX Lab # 1 Discussion How To Use The Spectrophotometer How To Use The Temperature Probe How To Use The Hot Plate Start Of Lab # 1 Exercise Links To Charts And Graphs

20 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB 1) Hypothesis 2) Data Table 3) Graph (mM Product vs Temperature) 4) Conclusion 5) Answers to questions 1 - 4 The following is what you should make sure that you include in your lab report

21 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Changes in temperature can dramatically influence the activity of most enzymes by affecting enzyme structure As temperatures increase enzyme activity is greater due largely to the increase in kinetic energy (energy of motion) that allows the enzyme and substrate to collide with higher frequency

22 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Once the temperature optimum is reached, enzyme activity decreases slowly as the enzyme is heated because the enzyme is undergoing denaturation Denaturation is a term used to describe a change in shape occurring to enzymes

23 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB When an enzyme’s shape is changed it can no longer perform its function as efficiently Just like a wrench that has lost its shape when over-heated in a forge, an enzyme is not able to function when it has lost its shape

24 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB This exercise is designed to help you understand the effect of temperature on enzyme activity Invertase You will also analyze data from this experiment to determine the ideal temperature optimum for Invertase (an enzyme) activity

25 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB To begin the experiment we will determine the temperature of the substrate  The substrate is the chemical material which the enzyme will be acting on

26 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB As is the case with real experiments in a biochemistry lab, enzyme should always be added to the substrate last to prevent the reaction from starting before all necessary components have been added

27 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB The first thing you will do is to develop a hypothesis to predict the effect of an increase in temperature on invertase activity Enter your hypothesis in the first line of your lab report  A hypothesis is a best guess at what you think may be the results of the experiment Click To Go To Lab Report Hypothesis

28 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Sucrose The substrate used in this experiment is Sucrose Substrate concentration for sucrose for this experiment is 90 millimolar units (mM)

29 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Invertase The enzyme used in this experiment is Invertase Invertase will react with sucrose in a biochemical reaction that results in the formation of by-products sucrose (+ water) glucose + fructose Invertase Sucrase

30 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Accumulation of by-products in the solution over time will create a change in the relative density of the solution Spectrophotometer This relative density can be measured by the use of a Spectrophotometer Photons 4000 ON

31 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB HOW TO USE THE SPECTROPHOTOMETER Photons 4000 ON

32 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 ON The spectrophotometer is designed to measure the amount of light which is entering into the device The device records a measurement in photons which can be extrapolated into a measure of density Spectrophotometer

33 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show In the beakers below and to the left is a series of solutions with varying degrees of density Photons 4000 ON When an enzyme such as invertase is added to a sucrose solution the resulting enzymatic reactions cause an accumulation of by-product Spectrophotometer

34 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 ON This accumulation of product results in an increase in density of the solutions Spectrophotometer By placing the beakers in front of a Spectrophotometer we are able to measure the relative density of each solution Spectrophotometer

35 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 Spectrophotometer ON Click on the top beaker (lightest color) which appears to have the least density The beaker will now be placed in front of the spectrophotometer

36 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 3200 ON ON Turn the spectrophotometer on by clicking the ON button to measure the light intensity which is passing through the beaker The measurement you see on the readout from the Spectrophotometer is in photons and will need to be converted to relative density Spectrophotometer

37 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 9000 3200 ON GREEN PHOTONSDENSITY Click on the GREEN button below the table to go to the chart which will convert the measurement of PHOTONS into units of DENSITY DENSITY The DENSITY units can be correlated to mM of product produced and are the values which you will use to graph your results Click To Go To Conversion Chart Spectrophotometer

38 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 Spectrophotometer ON Click on the second beaker which appears to have an intermediate density and then click the ON button on the spectrophotometer GREEN Use the conversion chart again to convert photons to density by clicking on the GREEN button below the table 2800 Click To Go To Conversion Chart

39 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4000 Spectrophotometer ON ON Click on the third beaker which appears to have the highest density and then click the ON button on the spectrophotometer GREEN Use the conversion chart again to convert photons to density by clicking on the GREEN button below the table 2600 Click To Go To Conversion Chart

40 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB HOW TO USE THE TEMPERATURE PROBE 35 o 30 o Temp

41 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Temp Hot Plate Temperature Probe Sucrose Solution A Temperature Probe can be used to measure the temperature of the sucrose solution A Temperature Probe is inserted into the solution which will give measurements of centigrade degrees

42 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC Temp Hot Plate Temperature Probe Hot Plate Click on the beaker containing the solution to set it on the Hot Plate Temperature Probe Now click on the Temperature Probe to insert it into the solution Sucrose Solution 30 o Record the measure of the temperature of the solution

43 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB HOW TO USE THE HOT PLATE HC

44 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Temp Hot Plate Temperature Probe Hot Plate A Hot Plate is used to heat the solution Hot Plate Temperature can be controlled by using the dial on the side of the Hot Plate Be very careful not to touch the Hot Plate with your hands or you may burn yourself Sucrose Solution

45 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 30 o 35 o Temp Temperature Probe Hot Plate Click the RED KNOB on the Hot Plate to increase the amount of heat Again insert the Temperature Probe into the solution by clicking on it and record the temperature Hot Plate

46 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Insert Temperature Probe by clicking on itTemp Photons 90003200 Turn Spectrophotometer On Hot Plate SpectrophotometerTemperature Probe Click red knob to start hot plate ON Familiarize yourself again with each of the instruments below Temperature ProbeSpectrophotometer Hot Plate Insert the Temperature Probe, Turn the Spectrophotometer on, and Turn up the heat on the Hot Plate for practice Return to Index

47 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Temp Photons 90004300 ON You are now ready to begin the experiment to see how temperature effects the rate of enzyme action Click on the Yellow button below to go to experiment #1 Click To Go To Experiment #1

48 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show In the beaker below is a 90 mM sucrose solution which will act as the substrate The temperature probe will be used to measure the temperature of the substrate solution before heating it with the Hot Plate Click on the temperature probe to measure the temperature of the substrate and then record your results in the data table under temperature for measurement #1 30 o Temp Click To Go To Data Table Measurement # 1

49 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Spectrophotometer Spectrophotometer Click on the beaker with the 90 mM sucrose solution to place it in front of the Spectrophotometer Spectrophotomete Next turn the Spectrophotometer on and record your data on your data sheet Photons ON 3600 Measurement # 1 Click To Go To Conversion Chart

50 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB At this point you should have recorded the first line of data on your data sheet for experiment #1 Data Table MeasurementTemperaturePhotonsmM of Product 13036000 2 3 4 5 6 7 8 9 10 11

51 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Next we will add an enzyme to the substrate solution (Sucrose) The enzyme we will use is called INVERTASE Invertase is an enzyme which will metabolize or break down sucrose When sucrose is broken down the build up of metabolic by-products causes changes in the density of the sucrose-invertase solution Sucrose solution Invertase

52 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show In the beaker below is a 90 mM sucrose solution test tube In the test tube to the right is an enzyme called invertase test tube Pour the invertase into the sucrose solution by clicking on the test tube

53 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Now that the enzyme has been added we will take the Sucrose- Invertase solution to our lab equipment to measure the effects of increasing temperatures on enzyme activity Sucrose- Invertase solution You will have to be quick to limit the effects of time on the actions of the enzyme with the substrate

54 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 3600 3200 ON HC 30 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record data Turn Spectrophotometer ON and record data Click red knob to start hot plate 35 o Measurement # 2 Click To Go To Next Measurement

55 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 3200 2800 ON HC 35 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 40 o Measurement # 3 Click To Go To Next Measurement

56 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 2800 2200 ON HC 40 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 45 o Measurement # 4 Click To Go To Next Measurement

57 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 2200 1400 ON HC 45 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 50 o Measurement # 5 Click To Go To Next Measurement

58 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 1400 700 ON HC 50 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 55 o Measurement # 6 Click To Go To Next Measurement

59 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 700200 ON HC 55 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 60 o Measurement # 7 Click To Go To Next Measurement

60 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 200 100 ON HC 60 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 65 o Measurement # 8 Click To Go To Next Measurement

61 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 100200 ON HC 65 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 70 o Measurement # 9 Click To Go To Next Measurement

62 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 200 400 ON HC 70 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 75 o Measurement # 10 Click To Go To Next Measurement

63 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 400 800 ON HC 75 o Temp Proceed to increase the temperature of the solution in 5 degree increments by clicking on the red knob of the hot plate Next insert the temperature probe record the temperature and then turn the spectrophotometer on and record the new reading Insert Temperature Probe by clicking on it and record dataTurn Spectrophotometer ON and record data Click red knob to start hot plate 80 o Measurement # 11 Click To Go To Next Measurement

64 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB At this point you should have recorded data for 11 measurements on your data table Next we will need to create a graph from the collected data which will help in interpreting the results

65 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB We will create data points on a graph in which we compare mM of product vs Temperature Use the Blue button below to view a graph where you will plot your data points  You should print this graph Click to View Graph Click to View Graph

66 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB After you have plotted all of your data points, connect all of the points by drawing a line from one data point to the next Your graph should look very similar to the graph shown from the link below Click to View Graph

67 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB From the graph that you have produced create a conclusion to the experiment Did your hypothesis agree with your results Click to View Conclusion Page

68 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB 1) What is the optimum temperature for invertase activity? 2) Explain why temperatures lower or higher than the optimum cause decreases in invertase activity? 3) Do you think that if a different substrate like glucose was used instead of sucrose, the results would be different? Why or why not? 4) Do you think that if a different enzyme like peptase was used instead of invertase, the results would be different? Why of why not? Answer the following questions and include them in your lab report Print this page

69 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB 1) Hypothesis 2) Data Table 3) Graph (mM Product vs Temperature) 4) Conclusion 5) Answers to questions 1 - 4 The following is what you should make sure that you include in your lab report

70 mM Product vs Temperature Mm of Product Temperature in degrees Celsius 3035404550556065707580 0 1 2 3 4 5 6 7 8 9 10 Click to Return to Last Slide Viewed

71 ENZYME LAB REPORT EXPERIMENT #1 Hypothesis: Click To Go To Last Viewed Slide Print this page

72 ENZYME LAB REPORT EXPERIMENT #1 Conclusion: Click To Go To Last Viewed Slide Print this page

73 ENZYME LAB REPORT Click To Go To Last Viewed Slide Experiment #1 Data Table MeasurementTempPhotonsmM of Product 1 2 3 4 5 6 7 8 9 10 11 Print this page

74 Photons % Density % Density mM of Product 360000 340050.5 3200101.0 3000151.5 2800202.0 2600252.5 2400303.0 2200353.5 2000404.0 1800454.5 1600505.0 1400555.5 1200606.0 1000656.5 800707.0 600757.5 400808.0 200858.5 0909.0 PHOTON TO mM of PRODUCT CONVERSION CHART Click To Go To The Last Slide Viewed Print this page

75 mM Product vs Temperature mM of Product Temperature in degrees Celsius 3035404550556065707580 0 1 2 3 4 5 6 7 8 9 10 Print this page Click to Return to Last Slide Viewed

76 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB #2 pH Influences On Enzyme Activity

77 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB #2 INDEX Lab # 2 Discussion How To Use The Spectrophotometer How To Use The pH Probe How To Use The Eye Dropper Start Of Lab # 2 Exercise Links To Charts And Graphs

78 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB 1) Hypothesis 2) Data Table 3) Graph (mM Product vs pH) 4) Conclusion 5) Answers to questions 1 - 4 The following is what you should make sure that you include in your lab report

79 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Another factor besides temperature that strongly influences enzyme activity in living cells is the pH of the environment in which the enzyme is designed to function Changes in pH can cause denaturation of enzymes thus reducing their activity

80 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB For example, in humans, a cytoplasmic protein in a skin cell is surrounded by a different fluid environment at a different pH than a membrane-bound enzyme like Invertase with an active site that projects out into the lumen of the small intestine

81 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB This exercise is designed to help you understand the effect of pH on enzyme activity by studying Invertase activity over a range of different pH values from acidic to basic conditions

82 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB To begin the experiment we will determine the pH of the substrate  The substrate is the chemical material which the enzyme will be acting on

83 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB As is the case with real experiments in a biochemistry lab, enzyme should always be added to the substrate last to prevent the reaction from starting before all necessary components have been added

84 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB The first thing you will do is to develop a hypothesis to predict the effect of changes in pH on Invertase activity Enter your hypothesis in the first line of your lab report  A hypothesis is a best guess at what you think may be the results of the experiment Click To Go To Lab Report Hypothesis

85 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Sucrose The substrate used in this experiment is Sucrose Substrate concentration for sucrose for this experiment is 90 millimolar units (mM)

86 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Invertase The enzyme used in this experiment is Invertase Invertase will react with sucrose in a biochemical reaction that results in the formation of by-products

87 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Accumulation of by-products in the solution over time will create a change in the relative density of the solution Spectrophotometer This relative density can be measured by the use of a Spectrophotometer Photons 4000 ON The use of the spectrophotometer should be familiar to you from Experiment #1Experiment #1

88 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB pH is a measure of the relative number of hydrogen and hydroxide ions present in a solution

89 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB The more hydrogen ions there are in a solution the more acidic the solution will act Less hydrogen ions and the more basic the solution will react H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ OH - H+H+ H+H+ H+H+ ACIDBASE

90 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB A device called a pH meter can be used to measure the relative number of hydrogen ions present in a solution pH of a particular solution is measured as an inverse exponential calculation of the number of hydrogen ions 35 o 7 pH

91 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB This means that the higher the number of hydrogen ions the lower the pH reading will be H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ OH - H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ pH = 6pH = 5

92 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB Thus the lower the pH the more acidic the solution is The pH scale ranges from a low of 1 to the highest pH of 14 1471

93 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB A pH measure of 7 indicates that the number of hydrogen ions and the number of hydroxide ions are equal and thus the solution will not express acidic or basic tendencies We would refer to solutions with a pH of 7 to be neutral 1471 Neutral

94 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB pH measuring less than 7 would be considered acidic pH measuring higher than 7 would be considered basic Low concentrations of hydrogen ions High concentrations of hydrogen ions 1471 BaseAcid Neutral

95 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB HOW TO USE THE pH PROBE 35 o 7 pH

96 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show A pH Probe can be used to measure the acidity of the sucrose solution A pH Probe is inserted into the solution which will give measurements of hydrogen ion concentrations 7 pH pH Meter pH Probe

97 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show To measure the pH of the solution click on the pH Probe The resulting value indicated on the pH meter can now be recorded 7 pH pH Meter pH Probe

98 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show In order to change the pH of the solution to measure enzyme activity we will add a prescribed amount of acid by using an eye dropper Below you will see a bottle of acid solution and an eye dropper HCl Acid Solution Sucrose Solution Eye Dropper

99 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB HOW TO USE THE Eye Dropper

100 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click on the eye dropper to sample a small amount of acid A small amount of acid is now present in the eye dropper Sucrose Solution Eye Dropper HCl Acid Solution

101 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click on the eye dropper again to present a single drop of acid to the sucrose solution We can now resample the sucrose solution for a change in pH Sucrose Solution HCl Acid Solution

102 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show To measure effect of pH on enzyme activity we will use a pH meter to measure acidity of the solution In order to measure the amount of product produced we will once again use a spectrophotometer as we had used in Experiment #1 Photons 4300 ON Spectrophotometer 7 pH pH Meter pH Probe Sucrose Solution

103 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 4300 ON You are now ready for experiment # 2 Sucrose Solution Eye Dropper You will be using the eye dropper to increase pH levels of the sucrose – invertase solution You will use the pH meter to measure pH and the spectrophotometer to measure enzyme activity 7 pH pH Meter Spectrophotometer Click To Go To Experiment #2 pH Probe Base Solution Acid Solution

104 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show We will be adding an enzyme to the substrate solution (Sucrose) The enzyme we will use is called INVERTASE Invertase is an enzyme which will metabolize or break down sucrose When sucrose is broken down the build up of metabolic by-products causes changes in the density of the sucrose-invertase solution Sucrose solution Invertase The substrate solution we will use is a 90 mM sucrose solution

105 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show test tube In the test tube to the right is an enzyme called invertase test tube Pour the invertase into the sucrose solution by clicking on the test tube Turn the timer on by clicking on the blue button on top of the timer and wait for 2 minutes While you are waiting you can check the pH by going to the next slide

106 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show 7 pH Insert the pH Probe by clicking on the probe Record the pH in your Data Table in the row for Measurement #1 When the timer has buzzed and stopped go to the next slide to test the solution for density Two Minutes have elapsed

107 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 1600 ON 7 pH Turn the spectrophotometer on and record the measurement in photons in your Data Table for Measurement #1 You will need to use a conversion chart to find the amount of product produced

108 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End ShowPhotons 1600 ON 7 pH GREEN You can find the conversion chart by using the GREEN button below At this point you will have completed one measurement for a single pH value Click to View Conversion Chart

109 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 2 We have now acquired a fresh solution of 90 mM sucrose Add one drop of acid to the sucrose solution by clicking on the eye dropper HCl Acid Solution

110 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 2 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

111 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #3 MEASUREMENT # 2 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 1200 ON 6 pH Two Minutes have elapsed

112 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB At this point your Data Sheet should look like below with two measurements evaluated Experiment #2 Data Table MeasurementpHPhotonsmM of Product 1716005 2612006 3 4 5 6 7 8 9 10 Click To Go To Measurement #3

113 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #3 We will now proceed with measurements 3 - 10 The procedure will be to: 1) add a measured amount of acid or base to the substrate 2) add enzyme to the substrate 3) start timer for substrate enzyme action 4) measure and record pH with pH meter 5) measure and record density with the spectrophotometer ON 6 pH Photons 4300

114 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 3 We have acquired a fresh solution of 90 mM sucrose Add two drops of acid to the sucrose solution by clicking on the eye dropper HCl Acid Solution

115 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 3 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

116 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #4 MEASUREMENT # 3 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 1000 ON 5 pH Two Minutes have elapsed

117 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 4 We have acquired another fresh solution of 90 mM sucrose Add four drops of acid to the sucrose solution by clicking on the eye dropper

118 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 4 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

119 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 4 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 1200 ON 4 pH Two Minutes have elapsed Click To Go To Measurement #5

120 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 5 We have acquired another fresh solution of 90 mM sucrose Add seven drops of acid to the sucrose solution by clicking on the eye dropper

121 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 5 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

122 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #6 MEASUREMENT # 5 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 1600 ON 3 pH Two Minutes have elapsed

123 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 6 We have acquired another fresh solution of 90 mM sucrose Add eleven drops of acid to the sucrose solution by clicking on the eye dropper

124 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 6 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

125 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #7 MEASUREMENT # 6 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 3200 ON 2 pH Two Minutes have elapsed

126 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 7 We have acquired another fresh solution of 90 mM sucrose Add nineteen drops of acid to the sucrose solution by clicking on the eye dropper

127 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 7 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

128 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #8 MEASUREMENT # 7 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 3600 ON 1 pH Two Minutes have elapsed

129 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 8 We have acquired another fresh solution of 90 mM sucrose Add one drop of base to the sucrose solution by clicking on the eye dropper NaOH Base

130 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 8 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

131 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #9 MEASUREMENT # 8 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 2400 ON 8 pH Two Minutes have elapsed

132 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 9 We have acquired another fresh solution of 90 mM sucrose Add five drops of base to the sucrose solution by clicking on the eye dropper NaOH Base

133 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 9 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

134 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click To Go To Measurement #10 MEASUREMENT # 9 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 2800 ON 9 pH Two Minutes have elapsed

135 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 10 We have acquired another fresh solution of 90 mM sucrose Add eleven drops of base to the sucrose solution by clicking on the eye dropper NaOH Base

136 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 10 Add the invertase enzyme by clicking on the test tube Now click on the timer and go to the next slide

137 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show MEASUREMENT # 10 Measure and record the pH of the solution by clicking on the pH probe When the timer has buzzed click on the spectrophotometer to measure and record the density Photons 3600 ON 10 pH Two Minutes have elapsed Click To Go To Next Slide You have finished collecting data for experiment #1

138 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB At this point you should have recorded data for 10 measurements on your data table Next we will need to create a graph from the collected data which will help in interpreting the results

139 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB We will create data points on a graph in which we compare mM of product vs pH Use the Blue button below to view a graph where you will plot your data points  You should print this graph Click to View Graph Click to View Graph

140 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB After you have plotted all of your data points, connect all of the points by drawing a line from one data point to the next Your graph should look very similar to the graph shown from the link below Click to View Graph

141 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB From the graph that you have produced create a conclusion to the experiment Did your hypothesis agree with your results Click to View Conclusion Page

142 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB 1) What is the optimum pH for invertase activity? 2) Explain why pH lower or higher than the optimum cause decreases in invertase activity 3) Do you think that if a different substrate like glucose was used instead of sucrose, the results would be different? Why or why not? 4) Do you think that if a different enzyme like peptase was used instead of invertase, the results would be different? Why of why not? Answer the following questions and include them in your lab report Print this page

143 ENZYME LAB 1) Hypothesis 2) Data Table 3) Graph (mM Product vs pH) 4) Conclusion 5) Answers to questions 1 - 4 The following is what you should make sure that you include in your lab report 142 Enzyme Lab Index Lab #1 Index Lab #2 Index Previous Slide Last Slide Viewed Lab Tutorial End Show

144 ENZYME LAB REPORT EXPERIMENT #2 Hypothesis: Print this page Click To Go To Last Viewed Slide

145 ENZYME LAB REPORT EXPERIMENT #2 Conclusion: Click To Go To Last Viewed Slide Print this page

146 ENZYME LAB REPORT Experiment #2 Data Table MeasurementpHPhotonsmM of Product 1 2 3 4 5 6 7 8 9 10 Print this page Click To Go To Last Viewed Slide

147 Photons % Density % Density mM of Product 360000 340050.5 3200101.0 3000151.5 2800202.0 2600252.5 2400303.0 2200353.5 2000404.0 1800454.5 1600505.0 1400555.5 1200606.0 1000656.5 800707.0 600757.5 400808.0 200858.5 0909.0 PHOTON TO mM of PRODUCT CONVERSION CHART Click To Go To The Last Slide Viewed Print this page

148 mM Product vs pH Mm of Product pH 12345678910 0 1 2 3 4 5 6 7 8 9 Click To Go To Last Viewed Slide Print this page

149 mM Product vs pH Mm of Product pH 12345678910 0 1 2 3 4 5 6 7 8 9 Click To Go To Last Viewed Slide

150 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME TUTORIAL HOW ENZYMES WORK 149 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

151 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Essential points about enzymes :  All enzymes are proteins  Each enzyme has a specific catalytic action  Their normal activity depends on their environment  Abnormal conditions cause reduced activity

152 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK

153 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Enzymes are globular proteins They have an area, usually thought of as a pocket-shaped gap in the molecule, which is called the active site

154 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Only the substrate fits into the active site The enzyme speeds up the process of conversion of substrates into products

155 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Although the enzyme obviously joins with the substrate for a short while, the enzyme and substrate split apart afterwards, releasing the enzyme Thus the enzyme is not used up in the process, so it can continue to react if more substrate is provided

156 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Within the normal range, changes in temperature, pH, and concentrations of substrate and enzyme affect the rate of reaction in accordance with predictable interactions between enzyme and substrate molecules

157 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK The effects of temperature may be explained on the basis of kinetic theory Increased temperature increases the speed of molecular movement and thus the chances of molecular collisions, so within a narrow range (often 0-45 °C), the rate of reaction is proportional to the temperature

158 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK It is often said that an enzyme's rate of reaction doubles for every 10° C rise in temperature denaturation However, the interaction between this positive effect of increased temperature and the negative effect of denaturation results in a different situation  Enzymes may be said to have an optimum temperature for their action

159 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Changes in the pH probably affect the attraction between the substrate and enzyme, and thus the efficiency of the conversion process Often, there is an optimum pH  Near to pH 7 (neutral)

160 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Above normal temperatures (60 °C), heat alters irreversibly the enzyme molecule This denaturation is due to molecular vibrations (caused by heat) which change the shape of the protein, altering the folding and internal cross-linkages in its polypeptide chains

161 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK These changes, especially in the region of the active site, mean that the enzyme is inactivated, even when returned to normal temperature It would be wrong to say that an enzyme is KILLED by heat, since it is only a molecule, not an organism

162 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK The higher the temperature to which the enzyme is subjected and the longer the heating is continued, the greater the proportion of damaged enzyme molecules and the result is that the conversion process becomes less and less efficient

163 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HOW ENZYMES WORK Below normal temperatures, enzymes become less and less active, due to reductions in speed of molecular movement, but this is reversible, so enzymes work effectively when returned to normal temperature

164 HOW ENZYMES WORK Enzymes are sometimes adversely affected by other chemical substances which combine with them, either at their active site or by altering the overall shape of their molecule Many drugs, poisons and pH changes have their effect in this way Enzyme Tutorial is Over Return to Index

165 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Extra slides

166 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show ENZYME LAB #1 INDEX Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show

167 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Mm of Product Temperature in degrees Celsius 3035404550556065707580 0 1 2 3 4 5 6 7 8 9 10

168 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Mm of Product Temperature in degrees Celsius 3035404550556065707580 0 1 2 3 4 5 6 7 8 9 10

169 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Click to add 5 0 35 o 40 o 45 o 50 o mM Product Temperature 35 o 40 o 45 o 50 o

170 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Insert Temperature Probe by clicking on itTemp Photons 45004300 Turn Spectrophotometer On Hot Plate Spectrophotometer Temperature Probe Click red knob to start hot plate ON

171 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC 35 o 30 o Insert Temperature Probe by clicking on itTemp Photons 45004300 Turn Spectrophotometer On Hot Plate Spectrophotometer Temperature Probe Click red knob to start hot plate ON

172 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Mm of Product Temperature in degrees Celsius 3035404550556065707580 0 1 2 3 4 5 6 7 8 9 10

173 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show Extra slides

174 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show mM Product vs pH Mm of Product pH 1234567891011 0 1 2 3 4 5 6 7 8 9 10

175 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show HC Temp Hot Plate Temperature Probe Hot Plate Click on the beaker containing the solution to set it on the Hot Plate Temperature Probe Now click on the Temperature Probe to insert it into the solution Sucrose Solution 30 o Record the measure of the temperature of the solution Click To Go To Next Slide

176 Enzyme Lab Index Lab #1 Index Lab #2 Index Next Slide Previous Slide Last Slide Viewed Lab Tutorial End Show 7 Insert pH Probe by clicking on itpH Photons 4300 Turn Spectrophotometer On pH Meter Spectrophotometer pH Probe Click eye dropper to add acid ON


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