Effects of pH and temperature on amylase activity

Aim: To find out the effects pH and temperature have on amylase activity using
starch and amylase

Introduction
Amylase is an enzyme that results in conversion of starch into sugars with the help
of suitable catalyst. They are found in plants, animals and other microorganisms and
huge amount of amylase fond in germinating cereals and saliva of different animals
bodies (Battestin and Macedo, 2007). They are synthesis similar to the proteins
throughout the ribosome present within the cells and react to the targeted substrates.
Due to that very reasons after conversion of colour from green to brown and
breakdown of starch results in sweeter taste of fruits such as bananas.

Method

To investigate the temperature first of all 5Cm 3 starch solution divided into 5 test
tubes and kept in a particular temperature of 0, 30, 40, 50 and 70 degree centigrade
respectively. After that another 0.53 cm 3 and starch and amylase were added at each
temperature and clock has been started. After every two minutes each solution
testing has been carried out and the process wee reputed continuously till the starch
elimination from the solution and iodine remains a brown colour.
Ph experiment has been carried out under controlled environment and 5 drops of
acid has been added to the first tube and 15 drops to the second test tubes and ph
was noted. And in test tube 4 5 drops of alkali added and in test tube 5 15 drops of

alkali added and ph measured accordingly and to each of the tube 0.5 cm 3 amylase
solution added for starch test (Effects of Cooked Starches and Sucrose or their
Combination on Salivary ?-Amylase Activity and Oral pH, 2016).

Results
Effect of temperature (table 1)
Temperature
0
30
40
50
70

Time (t) min

1.00
11.0
6.00

1/t (min-1)
0
1.000
0.092
0.168
0

It is evident from the above result that in extreme temperature native folded structure
of protein result in uncoils into random configuration and for this biological activity of
protein is diminished. The graph display reciprocal of thee against temperature
based on the data from table 1.

Figure1. Plot of the reciprocal of time against temperature for the enzymatic activity
Effect of pH (table 2)
pH
4
6

Time (t) min

11.0

1/t (min-1)
0
0.092

7
9
10

18.0
10.2
6.3

0.053
0.120
0.168

Table two shows reciprocal of time against ph based on data

The bell shaped curve is displayed and the longest pick indicates the optimum pH for
enzymatic activity.
Conclusion

The study provides a clear and concise idea regarding the analyse activity and the
effect of ph and temperature to the amylase activity are clearly decocted with the
help of the experiment and some direct and indirect correlations were spotted that
will help in further evaluation (Hashemi et al., 2013). Folded shape of enzymes
helps in better enzymatic action such as polypeptide chins are folded into three
dimensional shape of hydrogen binds and tertiary structure is effectively maintained
and the suitable shape help of the enzyme to develop active sites in which the
consequent substrate is bind and active site is different among each enzyme
depending upon the reaction process and properties.

Evaluation

The reaction rate can be increased if the pH remains constant as after collide ion
they enzyme do not bind to the substrate (Zhou and Chen, 2001). If higher
temperature is provided the substrate and enzyme molecule movement will be
increased consequently and it will also results in changing the shape of enzyme.

References
Battestin, V. and Macedo, G. (2007). Effects of temperature, pH and additives on the
activity of tannase produced by Paecilomyces variotii. Electron. J. Biotechnol.,
10(2), pp.0-0.
Effects of Cooked Starches and Sucrose or their Combination on Salivary ?-Amylase
Activity and Oral pH. (2016). IJSR, 5(2), pp.855-858.
Hashemi, M., Mousavi, S., Razavi, S. and Shojaosadati, S. (2013). Comparison of
submerged and solid state fermentation systems effects on the catalytic activity
of Bacillus sp. KR-8104 -amylase at different pH and temperatures. Industrial
Crops and Products, 43, pp.661-667.
Zhou, Q. and Chen, X. (2001). Effects of temperature and pH on the catalytic activity
of the immobilized -galactosidase from Kluyveromyces lactis. Biochemical
Engineering Journal, 9(1), pp.33-40.