ABSTRACT

The experiment is conducted to extract of commercially important enzyme from natural

sources besides to analyze the development of enzyme assays quantification of enzyme
activity and specific activity. Two sets of experiment are carried out one and these
experiments are used to first, for sample and another is for control. Aside from detecting the
color change on the sample-DNSA reagent mixture, the optical density (OD) reading is also
taken. After the centrifuge process the supernatant is taken to test for any enzymatic reaction
present. The supernatant is then mixed with starch solution again to test for enzymatic
activity. Then, two sets of experiment were prepared to test for the amylase activity, sample
and controls. The control set are let to boil in boiling water to deactivate the active site of the
enzyme thus no reaction will take place. Meanwhile the sample is let to settle in water bath of
37C to allow the optimum condition of enzyme activity. The results show that there is
positive reaction towards the DNSA solution of both the sample and control. It can be
concluded that optimal temperature for Bacillus Subtilis is at 37C as the growth of organism
inhibited with increasing in temperature. It is believed that the side reaction occurred in the
control set is what makes the DNSA mixture change it colour. While the brown colour
change of DNSA for the presence of reducing sugar from the initial yellow colour but the
colour strength are not being observed during the experiment. The results show that the
average reading for control set is 0.015 and the average OD reading for sample is 0.023. The
conclusion that can be made from the experiment is that there is an enzymatic activity and
the bacteria are capable of producing the enzyme. The objective is encountered and the
theory is accepted.

CONCLUSION

Amylase enzyme plays important role in biotechnological industries and has

several potential applications in food, fermentation, textile and paper industries. Amylase can
be obtained from several sources such as plants, animals, and microbes. The experiment is
conducted to extract of commercially important enzyme from natural sources besides to
analyze the development of enzyme assays quantification of enzyme activity and specific
activity. Two sets of experiment are carried out one and these experiments are used to first,
for sample and another is for control. Each set of experiment is running trice to get a better
insight on the results. Aside from detecting the color change on the sample-DNSA reagent
mixture, the optical density (OD) reading is also taken. After the centrifuge process the
supernatant is taken to test for any enzymatic reaction present it is also mixed with starch
solution again to test for enzymatic activity. Then, two sets of experiment were prepared to
test for the amylase activity, sample and controls. The sample is let to settle in water bath of
37C and it is proved that positive reaction towards the DNSA solution of both the sample
and control.
It can be concluded that optimal temperature for Bacillus Subtilis is at 37C as the
growth of organism inhibited with increasing in temperature. It is believed that the side
reaction occurred in the control set is what makes the DNSA mixture change it colour. While,
brown colour change of DNSA for the presence of reducing sugar from the initial yellow
colour but the colour strength are not being observed during the experiment due to human
error during the experiment is carried out. As for the OD reading for each set of the
experiment, the results show that the average reading for control set is 0.015 and the average
OD reading for sample is 0.023. The conclusion that can be made from the experiment is that
there is an enzymatic activity and the bacteria are capable of producing the enzyme. The
objective is encountered and the theory is accepted.