Group 4: effect of AB dye concentration

Abs. t=0 Abs. t=10 min

% Dye
[AB] ppm
Degradation
Ao Af
500 0.596 0.580 2.684

1000 1.472 1.427 3.057

2000 2.285 2.181 4.551

3000 0.977 0.946 3.173

In this part, we studied the effect of the substrate concentration on the enzyme activity. Due to increased
collisions between the substrate and the enzyme, the rate of the reaction increases as the substrate
concentration does. As noticed in the table, the %degradation increases with increasing amide black dye
concentration from 500 ppm until reaching the optimum value which was 2000 ppm where the
%degradation was equal to 4.551%. The enzyme can tolerate high concentration of AB dye,until 2000
ppm. The lower % degradation was at concentration 500 which was 2.684%. Then comes the %
degradation at concentration 1000 which was 3.057%. Then comes the %degradation at concentration of
3000 which was 3.173%. The rate is constant at increased substrate concentrations. The enzyme active
site is fully saturated (occupied) with substrate such that adding more substrate does not increase the rate
of the reaction.

Group5: effect of SBP enzyme concentration

Abs. t=0 Abs. t=10 min % Dye

[SBP] µM
Ao Af Degradation
0 2.245 2.174 3.16

0.26 µM 2.345 2.232 4.82

2.6 µM 2.237 1.754 21.59

26 µM 2.381 0.426 82.11

In this part of the experiment, we studied the effect of the concentration of the enzyme on the reaction. As
observed from the table, the rate of the reaction increases, when the concentration of SBP enzyme
increases. The % degradation at concentration 0 was 3.16 % which was the lowest %degradation. The %
degradation of 0.26μM was 64.82%. Moreover the % degradation at concentration 2.6μM was 21.59 %.
Finally, the optimum value was at concentration 26μM, which was 82.11%. Because there are more
enzymes present to support the breakdown of the substrate, concentration enhances reactivity and
produces higher degradation. However, a point will be reached where no matter how much enzyme is
presented, the reaction rate will be constant, and this is equilibrium. This happens when all the substrates
are broken down by the exact same amount of enzyme, so any excess enzyme presented will not have any
additional substrate to break it down. So briefly The enzyme can tolerate high concentration SBP dye,
until 26 µM. The control here at 26 µM has the highest concentration while the percentage of the other
increases with increasing concentration.

Group6: effect of denaturants

Abs. t=0 Abs. t=10 min % Dye

Denaturants
Ao Af Degradation
0 3.065 1.674 50.24

1.5 % SDS 3.157 1.594 49.5

5 % SDS 3.119 1.552 45.38

6 M HCl 3.152 2.266 28.11

In this experiment, we studied the effect of denaturants of the reaction rate. Degradation was predicted to
reduce by adding denaturants. Denaturants can inhibit enzyme activity and stop its catalytic ability, which
slows down the rate of the reaction when they are added. From the obtained results, the 1.5% SDS didn't
affect the degradation much where degradation was 49.5%, and at 5% SDS it decreased to 45.38%, while
the 6M of HCl decreased the degradation to 28.11% since HCl is a denaturation reagent that will denature
the enzyme. The optimum value is obtained when there's no denaturant added to the reaction. The
percentage of degradation in the optimum is 50.24% which was the highest %degradation. By the
addition of different types of denaturants, this percentage decreases. Detergents can generally be tolerated
by the enzyme depending on their type and concentration when administered. So briefly the enzyme can’t
tolerate high concentration of the detergent. The enzyme can tolerate detergent depending on the type and
concentration of the detergent that is added.

Discussion:
In conclusion, the main objective of this experiment was fully achieved as we studied the effect of
temperature, pH, Inhibitors, concentration of substrate and enzyme and denaturants on enzyme activity.
Enzymes are part of our daily lives. Metabolism of the food we eat is performed by enzymes. In
detergents, enzymes are used to clean stains found on clothes. Sewage and industrial wastes are treated
with enzymes to make them less toxic to humans and to the environment by making them less pollutant.
At the end of this experiment, we were able to reach most of the expected results. Some errors may have
occurred due to technical errors, in addition to some human errors. Further experimenting is
recommended to examine the results and indeed increase precision and accuracy of the obtained results.