04_Enzyme+Action+experiment

 ENZYME ACTION! = Aim   =  The aim of this experiment is to investigate the effect of temperature and pH on enzyme action.  = Hypothesis   =  If the temperature is too high or low away from the body temperature (around 37°C), there would be no enzyme action. If the = Apparatus/ Materials   = o  Bunsen burner, tripod stand and gauze o  3 water baths o  Thermometer o  12 test tubes and rack o  Measuring cylinder o  4 250ml beakers o  2 white tiles o  2 droppers o  Ice cubes o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> Distilled water o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> Test tube holder o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> Stop watch o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> Goggles o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> 2% starch solution o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> 1% amylase solution o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> 1 M Hydrochloric acid solution o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> 1 M Ammonium Hydroxide solution o <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> Iodine solution =<span style="font-family: 'Spranq eco sans','sans-serif';"> Methodology   = <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> Part A- How temperature affects enzyme reaction <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 1. Label 8 test tubes A1, A2, B1, B2, C1, C2, D1 and D2. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 2. Add 5cm³ of starch solution into tubes A2, B2 and C2. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 3. Add 3 cm³ of amylase solution into tubes A2, B2 and C2. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 4. Place A1 and A2 into a beaker of ice water. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 5. Place B1 and B2 into a water bath at 37°C (body temperature. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 6.  Place D1 and D2 into another water bath at 37°C.  <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 7.  Place C1 and C2 into boiling water.  <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 8.  Leave the tubes to stand for 10 minutes.  <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 9.  Pour the contents of A2, B2, C2, and D2 into the starch tube, A1, B1, C1 and D1. Record the time IMMEDIATELY. Keep tubes A1, B1, C1 and D1, which contains the mixtures in the respective water baths throughout the experiment.  <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 10.  After 10 minutes, test the mixture in each tube with iodine solution, on a white tile. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 11. Record observations <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> Part B- How pH level affects enzyme reaction <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 1. Add 2cm³ of starch solution to each of the test tubes, W, X, Y, Z. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 2. Fill test tube W with 2 cm³ of HCL and 2 cm³ of amylase. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 3. Fill test tube X with 2 cm³ of distilled water and 2 cm³ of amylase. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 4. Fill test tube Y with 2 cm³ of NH <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%; position: relative; top: 5.5pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> OH and 2 cm³ of amylase. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 5. Fill test tube Z with 4 cm³ of distilled water. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 6. Place the tubes in a water bath at 37°C. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 7. Test the solution with iodine solution. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 8. Time for 10 minutes. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 9. After 10 minutes, test the solution with iodine solution again. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> 10. Record observations <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;"> =<span style="font-family: 'Spranq eco sans','sans-serif';"> Results = =<span style="font-family: 'Spranq eco sans','sans-serif';"> = <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Part A <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Part B =<span style="font-family: 'Spranq eco sans','sans-serif';"> = =<span style="font-family: 'Spranq eco sans','sans-serif';"> = =<span style="font-family: 'Spranq eco sans','sans-serif';"> Discussion = <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">1. Which tube shows that starch digestion has occurred? Give your reasons. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Tube B shows that starch digestion has occurred. This is because starch was not present when it was tested for starch using the iodine solution. Therefore, the initial amount of starch has been digested.
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">Test Tube ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Contains ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Presence of starch ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Test Tube ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Contains ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Temperature (°C) ** ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">A1 ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">5cm³ starch solution <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">Yes  || **<span style="font-family: 'Spranq eco sans','sans-serif';">A2 ** || <span style="font-family: 'Spranq eco sans','sans-serif';">3cm <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">³ amylase solution <span style="font-family: 'Spranq eco sans','sans-serif';">  || <span style="font-family: 'Spranq eco sans','sans-serif';">0  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">B1 ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">5cm³ starch solution <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">No  || **<span style="font-family: 'Spranq eco sans','sans-serif';">B2 ** || <span style="font-family: 'Spranq eco sans','sans-serif';">3cm <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">³ amylase solution <span style="font-family: 'Spranq eco sans','sans-serif';">  || <span style="font-family: 'Spranq eco sans','sans-serif';">37  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">C1 ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">5cm³ starch solution <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">Yes  || **<span style="font-family: 'Spranq eco sans','sans-serif';">C2 ** || <span style="font-family: 'Spranq eco sans','sans-serif';">3cm <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">³ amylase solution <span style="font-family: 'Spranq eco sans','sans-serif';">  || <span style="font-family: 'Spranq eco sans','sans-serif';">100  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">D1 ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">5cm³ starch solution <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">Yes  || **<span style="font-family: 'Spranq eco sans','sans-serif';">D2 ** || <span style="font-family: 'Spranq eco sans','sans-serif';">3cm <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">³ distilled water <span style="font-family: 'Spranq eco sans','sans-serif';">  || <span style="font-family: 'Spranq eco sans','sans-serif';">37  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">Test Tube ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Contains ** || **<span style="font-family: 'Spranq eco sans','sans-serif';">Iodine Test ** ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">W ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">2cm³ of starch solution + 2 cm³ of HCL + 2 cm³ of amylase <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">Yes  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">X ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">2cm³ of starch solution + 2 cm³ of distilled water + 2 cm³ of amylase <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">No  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">Y ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">2cm³ of starch solution + 2 cm³ of NH <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%; position: relative; top: 5.5pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">OH + 2 cm³ of amylase <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">No  ||
 * **<span style="font-family: 'Spranq eco sans','sans-serif';">Z ** || <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt;">2cm³ of starch solution + 4 cm³ of distilled water <span style="font-family: 'Spranq eco sans','sans-serif';"> || <span style="font-family: 'Spranq eco sans','sans-serif';">Yes  ||
 * __<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Part A: __**

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">2. Which of the test tubes contained mixtures that gave a blue-black colour?

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">3. Why did the mixtures in tubes stated in (2) remain blue-black?

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">4. What purpose does the tube D1 serve?

It acts as the control for the experiment.

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">5. Using your knowledge of the structure of starch, describe the test you would use to detect the presence of the substance produced as a result of the reaction in test tube stated in (1) above?

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">6. What conclusion(s), based on the observations in this experiment, can be made about the effect of temperature on enzyme action?

Body temperature is the optimum temperature for enzyme action to take place. Temperatures that are much higher or lower than that of a average body temperature does not allow enzyme action to take place. This can be seen from the fact that test tube B does not have presence of starch where as test tubes A and C do. The main difference between these three test tubes is their temperatures, which are 37°C, 0°C and 100°C respectively. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">1. Which tube shows that starch digestion has occurred? Give your reasons. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Tube X and Y. <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">2. Which of the test tubes contain solutions that gave a blue-black colour?
 * __<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">Part B: __**

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">3. Why did the mixtures in the tubes stated in (2) remain blue-black?

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">4. What conclusion(s) based on the observations in this experiment, can be made about the effect of pH on enzyme action?

The pH level does effect the enzyme action. If the pH level is too low, enzyme action cannot take place. This can be seen from the observation that test tube W had starch present whereas tube Y had no starch present. The two tubes had similar contents with the main difference being that tube W contains an acid, HCl which pH level is less than 7, whereas tube Y contains an alkali, NH <span style="font-family: 'Calibri','sans-serif'; font-size: 11pt; line-height: 115%; position: relative; top: 5.5pt;"> <span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">OH which has a pH level higher than 7. Therefore the if the pH level is too low then enzyme action cannot take place.

<span style="font-family: 'Spranq eco sans','sans-serif'; font-size: 12pt; line-height: 115%;">5. How does temperature and pH affect enzyme action?

Body temperature (37°C) is the optimum temperature for enzyme action to take place. Temperatures that are much higher or lower than that of a average body temperature does not allow enzyme action to take place. If the pH level is too low, enzyme action cannot take place.