Beruflich Dokumente
Kultur Dokumente
6, © March 4, 2006
3nd Quarter A.Y. 2005-2006
ABSTRACT
Enzymes are special proteins that function to were computed using Lineweaver-Burke plot
lower the activation energy required for a and the Hanes-Woolf plot which was 2.116x10-05
reaction to occur, thus increasing the rate of mol/mL and 0.139 mL/s. The Eadie-Hofstee plot
reaction. One such enzyme studied in this was omitted since the actual plot varies from the
experiment is catalase. Catalase was extracted theoretical plot. From these parameters, the
from unripped papaya. Only the frist trial were enyme activity computed was 1313.3980 mol
used in the analysis since the second trial wasn’t H2O2 / sec · mol.
performed properly and increased in error might
occur. Five set-ups were made with varying Keywords: emzyme, catalase, activity, hydrogen
hydrogen peroxide and phosphate buffer peroxide, phosphate buffer.
solutions to obtain a relationship. Km and Vmax
______________________________________________________________________________________
50
Volume (mL) 40 A
Set-up A B C D E B
30
Sample extract 5 5 5 5 5
VO2
C
Phosphate buffer 0.8 0.6 0.4 0.2 -- 20 D
30% H2O2 0.2 0.4 0.6 0.8 1 10 E
0
0 100 200 300 400 500 600 700
Each of the flasks was immediately -10
Time
sealed. The amount of water displaced by the
oxygen gas was monitored. Volume readings
were listed in uniform intervals of 10 seconds.
Two trials were made.
The Lineweaver-Burke and Eadie-
Data Analysis Hofstee plot was made and both graph obtained a
Km value of 2.116x10-05 mol/mL and a Vmax of
Determination of Initial Velocity 0.139 mL/s.
For each reaction flask VO2 vs. time
were plotted. Velocity was calculated by taking Lineweaver-Burke
15
10
of O2 produced, the pressure of dry O2, and 0 10000 20000 30000 40000
1/[S]
50000 60000 70000 80000
0.25
0.2
0.15
v
0.1
0.05
0
0 2000 4000 6000 8000 10000 12000
v/[S]
A B C D E
Time (s)
ht (mm) Vol (mL) ht (mm) Vol (mL) ht (mm) Vol (mL) ht (mm) Vol (mL) ht (mm) Vol (mL)
10 0 0 0 0 0 0 0 0 0 0
20 0 0 4 2.22 0 0 1 0.57 7 3.89
30 0 0 9 5 0 0 3 1.67 17 9.44
40 2 1.11 12 6.67 0 0 6 3.33 22 12.22
50 3 1.67 13 7.22 2 1.11 7 3.89 26 14.44
60 5 2.78 15 8.33 5 2.78 9 5 34 18.89
70 6 3.33 16 8.89 8 4.44 10 5.56 34 18.89
80 7 3.89 17 9.44 11 6.11 11 6.11 34 18.89
90 8 4.44 19 10.56 13 7.22 12 6.67 36 20
100 8 4.44 20 11.11 14 7.79 14 7.78 38 21.11
110 9 5 21 11.67 16 8.89 15 8.33 40 22.22
120 10 5.56 21 11.67 16 8.89 16 8.89 42 23.33
130 10 5.56 23 12.79 16 8.89 18 10 45 25
140 11 6.11 24 13.33 16 8.89 19 10.56 47 26.11
150 11 6.11 24 13.33 17 9.44 20 11.11 48 26.67
160 11 6.11 25 13.89 17 9.44 21 11.67 50 27.78
170 11 6.11 26 14.44 17 9.44 22 12.22 52 28.89
180 12 6.67 26 14.44 17 9.44 24 13.33 53 29.44
190 12 6.67 27 15 17 9.44 25 13.89 55 30.56
200 12 6.67 27 15 17 9.44 26 14.44 57 31.67
210 13 7.22 28 15.56 17 9.44 27 15 59 32.79
220 13 7.22 28 15.56 17 9.44 28 15.56 60 33.33
230 13 7.22 28 15.56 17 9.44 29 16.11 62 34.44
240 13 7.22 29 16.11 17 9.44 30 16.67 63 35
250 13 7.22 29 16.11 17 9.44 31 17.22 64 35.56
260 13 7.22 29 16.11 17 9.44 32 17.78 67 37.22
270 13 7.22 30 16.67 34 18.89 68 37.78
280 14 7.79 30 16.67 36 20 70 38.89
290 14 7.79 30 16.67 37 20.56 71 39.44
300 14 7.79 30 16.67 38 21.11 72 40
310 15 8.33 30 16.67 38 21.11 73 40.56
320 15 8.33 31 17.22 38 21.11 74 41.11
330 15 8.33 31 17.22 38 21.11 75 41.67
340 15 8.33 31 17.22 38 21.11 76 42.22
350 15 8.33 38 21.11 78 43.33
360 15 8.33 39 21.67 79 43.89
370 16 8.89 39 21.67 81 45
380 16 8.89 39 21.67 82 45.56
390 39 21.67 83 46.11
400 40 22.22 84 46.67
410 40 22.22 85 47.22
420 40 22.22 85 47.22
430 41 22.78 86 47.78
440 41 22.78 87 48.33
450 42 23.33 88 48.89
460 42 23.33 88 48.89
470 42 23.33 89 49.44
480 43 23.89 90 50
490 43 23.89 90 50
500 43 23.89 91 50.56
510 43 23.89 91 50.56
520 44 24.44 92 51.11
530 44 24.44 92 51.11
540 93 51.67
550 93 51.67
560 93 51.67
570 94 52.22
580 95 52.78
590 95 52.78
Velocity (mL/s) Height (mm) PO2 (mmHg) PO2 (atm) VO2 (mL) V (L) Mole O2 [H2O2], mol/mL
A 0.0556 16 737.4065 0.9703 8.88 0.0089 3.5238x10-04 1.4095x10-05
B 0.2278 31 738.5094 0.9717 17.22 0.0172 6.8435x10-04 2.7374x10-05
C 0.1667 17 737.48 0.9704 9.44 0.0094 3.7464x10-04 1.4986x10-05
D 0.0556 44 739.4653 0.9730 24.44 0.0244 9.7255x10-04 3.8902x10-05
E 0.1111 95 743.2153 0.9779 52.78 0.0528 2.1109x10-03 8.4438x10-05
CONCLUSION
LITERATURE CITED
3.http://www.lsbu.ac.uk/biology/enzyme/practica
l1.html