Biochem lab report kinetics

Pablo Cuate
Biochemistry Lab Report

Objective: The purpose of this lab is to analyze enzyme activity at pH neutral for acid phosphatase as the colorless p-nitrophenylphosphate (PNPP) substrate concentration changes. We will empirically determine the concentration of production formation, Vmax and Km from absorbance of yellow PNP in the reaction.

Results
Figure 1

This figure indicates a positive linear trend for the concentration of PNP and absorbance.
Table 1: Absorbance Value for Acid Phosphatase Reaction at Varying Concentrations of PNPP with the Final Product Concentration

Time(s)
5mM(PNPP)
Product (PNP)1
Product(PNP)1μM
2.5mM PNPP
Product (PNP)2
Product(PNP)2μM
1mM PNPP
Product (PNP)3
Product(PNP)3μM
0

15

30
0.102
0.011
11.075
0.034
0.006
5.745
0.052
0.007
7.156
45
0.138
0.014
13.897
0.087
0.010
9.900
0.075
0.009
8.959
60
0.174
0.017
16.719
0.117
0.012
12.251
0.097
0.011
10.683
75
0.213
0.020
19.776
0.147
0.015
14.603
0.120
0.012
12.486
90
0.250
0.023
22.676
0.176
0.017
16.876
0.146
0.015
14.524
105
0.287
0.026
25.576
0.208
0.019
19.384
0.170
0.016
16.405
120
0.324
0.028
28.476
0.237
0.022
21.657
0.197
0.019
18.522
135
0.370
0.032
32.082
0.268
0.024
24.087
0.216
0.020
20.011
150
0.408
0.035
35.060
0.298
0.026
26.438
0.239
0.022
21.814

0.2mM PNPP
Product (PNP)4
Product(PNP)4μM
0.5mM PNPP
Product (PNP)5
Product(PNP)5μM
0.25mM PNPP
Product (PNP)6
Product(PNP)6μM

0.054
0.007
7.313
0.016
0.004
4.335
-0.033
0.001
1
0.078
0.009
9.194
0.038
0.006
6.059
-0.012
0.002
2.13983383
0.099
0.011
10.840
0.059
0.008
7.705
0.003
0.003
3.315566703
0.119
0.012
12.408
0.084
0.010
9.665
0.021
0.005
4.726446151
0.141
0.014
14.132
0.106
0.011
11.389
0.038
0.006
6.058943408
0.163
0.016
15.857
0.128
0.013
13.113
0.055
0.007
7.391440665
0.185
0.018
17.581
0.148
0.015
14.681
0.074
0.009
8.880702304
0.206
0.019
19.227
0.170
0.016
16.405
0.091
0.010
10.21319956
0.227
0.021
20.873
0.189
0.018
17.895