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



Difference02334566777



Table 2.2: Class Data on the Effect of Enzyme Concentration on Enzyme ActivityPart B:The Effect of Enzyme Concentration on Enzyme Activity


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


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







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


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


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


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Enzyme Lab 2