Download - Enzyme Lab 2
The Effect of Temperature, pH and Concentration on EnzymesIntroductionEnzymes are classified as highly specific proteins that function as catalysts working to facilitate biochemical reactions by lowering the activation energy needed for the reaction. Enzymes consist of two parts, the active site; which binds to the substrate forming an enzyme-substrate complex that aids in the chemical reaction, and the allosteric site; which links to specific cofactors forming the correct 3 dimensional shape. The ruction can be summarized with the equation :( Enzyme + Substrate Enzyme Substrate Complex Enzyme + Product), you can see by this equation that the enzymes are not consumed by the reaction; rather they are recycled and reused. Because these sites are very unique each individual enzyme requires highly specific terms and conditions of the reactions in which they will catalyze. Because of how specialized they are enzymes are also easily affected by external factors such as temperature pH and concentrations.Methodsa. Materials (for all procedures)Emma HansonMartinsonAdvanced Biology11-19-14
Hanson
50 mL beaker containing fresh catalase (yeast) solution Reaction chamber Ring stand Clamp 10mL graduated cylinder 100mL graduated cylinder 3% hydrogen Peroxide Pan (water bath) Pipette Hot plate Ice Thermometer Boiled catalase Buffers with pH: 4, 7 and 10 Distilled water Balance NaCL (salt)
b. Procedure Summary of general lab procedureBegin by using the water pan, ring stand, clamp, 100mL graduated cylinder and reaction chamber to create a setup as illustrated in the diagram below. Create a column of water in the graduated cylinder by submerging the cylinder in the water completely then flip it upside down and secure it to the ring stand being sure to keep the opening submerged. To test Part A; combine 1mL catalase solution with 10mL hydrogen in the reaction chamber and immediately seal the chamber with the stopper and hold under water. Measure the level of gas in the cylinder at 30 second intervals for 5 minutes. For Parts B, D, E and F repeat this test 3-4 times altering the enzyme concentration, pH, substrate concentration and ionic concentration for each respective test. For Part C repeat the test 3 times changing the temperature of the water for each test.
c. Statistical method RateThe rate of the reaction quantitatively measured by the difference in each consecutive recordings of the amount of water displaced in the graduated cylinder by the oxygen produced in the reaction.
To find to rate of reaction for each test take the average of the rates for all five intervals
Part A: Timea. Data table Team and Class dataTable 1.1: Team Data for the Time Course of Enzyme Activity using 3%Time 030sec1min1:302min2:303min3:304min4:305min
Difference 00001234666
Table 1.2: Class Data for the Time Course of Enzyme Activity using 3%The Time Course of Enzyme Activity using 3%
TimeKS,DH,JK,BEJO,JNAA,KS,OHSM,HB,RWJW,EH,ARMH,ND,MMAverage
00000000
30sec201.53.5011.333333
1min4025032.333333
1:30:005236053.5
2 min533.57174.416667
2:30:00644.57.5285.333333
3 min6468396
3:30:006479496.5
4 min648106107.333333
4:30:0075910.56118.083333
5min7610126128.833333
b. Graph Team and class data
Part B: Enzyme Concentrationa. Data table Team and Class dataTable 2.1: Team Data on the Effect of Enzyme Concentration on Enzyme ActivityTime 030sec1min1:302min2:303min3:304min4:305min
.75mL Concentration catalase
Difference 02223445789
.50mL Concentration catalase
Difference02334566777
.25mL Concentration catalase
Difference01333333444
Table 2.2: Class Data on the Effect of Enzyme Concentration on Enzyme ActivityPart B:The Effect of Enzyme Concentration on Enzyme Activity
.75mL Concentration catalase
TimeND MH MMJOJNKS,DH,JK,BEAA, KS, OHSM, HB, RWJW EH ARTotalClass average
000000000
3014.5224215.52.583333
13522.55219.53.25
1:30:005533.55223.53.916667
2554453264.333333
2:30:0055.5555429.54.916667
356.555.554315.166667
3:30:00575665345.666667
457.5667738.56.416667
4:30:0057.5667.58406.666667
5596689437.166667
.50 mL Concentration catalase
ND MH MMJOJNKS,DH,JK,BEAA, KS, OHSM, HB, RWJW EH ARTotalClass Average
000000000
303120.5129.51.5
14231.51.53152.5
1:30:00423323172.833333
24343.52420.53.416667
2:30:0044442.3523.33.883333
344.5452.56264.333333
3:30:0045462.5627.54.3
44546.53729.54.916667
4:30:0045.5473730.55.083333
545.5483.57325.333333
.25mL Concentration catalase
ND MH MMJOJNKS,DH,JK,BEAA, KS, OHSM, HB, RWJW EH ARTotalClass Average
000000000
3010302171.166667
120.5302310.51.75
1:30:0020.5302.53111.833333
2213033122
2:30:0021313.5313.52.25
32131.54.53152.5
3:30:0021324.5315.52.583333
421.5325417.52.916667
4:30:0021.5325417.52.916667
522325.5418.53.083333
b. Graph Team and Class data
c. Questions: # 7 parts a-cPart C: Temperaturea. Data tableTeam and Class dataTable 3: Class Data on the Effect of Temperature on Enzyme ActivityPart C:The Effect of Temperature on Enzyme Activity
5C37C100C
TimeSM, HB, RWJOJNTotalClass AverageSM, HB, RWJOJNTotalClass AverageSM, HB, RWJOJNTotalClass Average
0000000000000
301.501.50.759514720.52.51.25
1202112.5820.510.2520.52.51.25
1:302.313.31.6517.5825.512.7520.52.51.25
22.524.52.2522.5830.515.2520.52.51.25
2:30336324.51034.517.2520.52.51.25
33.536.53.2526.51036.518.2521.53.51.75
3:303.736.73.3529.51140.520.2521.53.51.75
44.537.53.7532.51143.521.7521.53.51.75
4:304.948.94.4534.51246.523.2521.53.51.75
55.349.34.6535.51247.523.7521.53.51.75
b. Graph Class data
c. Questions -- # 8 parts a-cPart D: pHa. Data table Team and Class dataTable 4.1 Team Data on the Effect of pH on Enzyme ActivityTime 030sec1min1:302min2:303min3:304min4:305min
.75mL Concentration catalase
Difference 03334456778
.50mL Concentration catalase
Difference01223455666
.25mL Concentration catalase
Difference02334444556
Table 4.2 Class Data on the Effect of pH on Enzyme ActivityPart D:The Effect of pH on Enzyme Activity
pH4710
TimeND MH MMAA,KS,OHJW AR EHAverageND MH MMAA,KS,OHJW AR EHAverageND MH MMAA, KS, OHJW EH ARAverage
0000000000000
303232.6666673111.6666675223
143.533.54222.66666752.533.5
1:3043.533.55323.33333353.533.833333
243.543.83333364.534.553.544.166667
2:30444465.545.1666676444.666667
344.554.56956.66666764.544.833333
3:3044.564.833333610576545
44575.33333361167.6666676555.333333
4:304575.333333612686555.333333
54585.66666761368.33333365.565.833333
b. Graph Team and class data
c. Questions --# 9 parts a-cPart E: Substrate concentrationa. Data table Team and Class dataPart E:The Effect of Substrate Concentration on Enzyme Activity
0%0.30%1.50%
TimeKS,DH,JK,BEAA, KS, OHMH, ND, MMAverageKS,JK,DH,BEAA, KS, OHMH, ND, MMAverageKS,JK,DH,BEAA, KS, OHMH. ND. MMAverage
0000000000000
3000003201.6666672222
10000330232.522.5
1:3000003402.3333334343.666667
2000034.502.543.543.833333
2:30000035.504.254444
3000045.503.16666754.544.5
3:30000045.503.1666675655.333333
4000045.524.756655.666667
4:30000045.523.8333336756
5000045.524.757756.333333
b. Graph Team and class data
c. Question -- # 10 parts a-cPart F: Ion concentrationa. Data table Team and Class dataTable 6.1 Team Data on The Effect of Ionic Concentration on Enzyme ActivityTime030sec1min1:302min2:303min3:304min4:305min
10% NaCl
Difference03333333.53.53.53.5
2% NaCl
Difference012233.53.54.54.55.55.5
0% NaCl
Difference0011.52345556
Table 6.2 Class Data on the Effect of Ionic Concentration on Enzyme ActivityPart F:The Effect of Ionic Concentration on Enzyme Activity
10% NaCl2%0%
TimeKS,JK,DH,BEJOJNSM, HB, RWAverageKS, JK, DH, BEJOJNSM, HB, RWAverageKS, JK, DH, BEJOJNSM, HB, RWAverage
0000000000000
3031531242.3333330332
13263.666667255413.532.5
1:30336425541.53.543
233.564.1666673554.33333323.553.5
2:303464.3333333.5554.533.553.833333
33464.3333333.5554.54464.666667
3:303.5564.8333334.5554.8333335465
43.5564.8333334.55.5555475.333333
4:303.5564.83333355.555.16666754.575.5
53.5564.83333355.555.16666764.596.5
b. Graph Team and class data