INTRODUCTION

Enzyme is a protein in the cell which lowers the activation energy of catalysed reaction, thus increasing
the rate of reaction. The action of enzyme is to increase the reaction of the biochemical conversion of
a substrate into something else. It can be simply described as follows:

The objectives of this experiments were to comprehend physical characteristics and properties of
enzymes also to study the factors affecting their activity. However, the parameter of this experiment
based on substrate concentration and on pH.

THEORY
Absorption photometry was preferred methods because of its simple technique and reliable. In this
experiment UV-visible spectrophotometer was used to determine the substrate concentration and
absorbance.

Figure 1 : Schematic diagram of UV- visible spectrophotometer

In absorption photometry the Bouguer- Lambert- Beer law, is applied for very dilute solutions. Where
the relationship exists between absorbance A vs concentration:

Io
absorbance, A = log = ε. c . d
I
A
and c= ε . d

Where: c is the concentration, ε is the absorption coefficient , d is the path length.

Enzyme kinetic is important in enzyme analysis in order both to understand the basic enzymatic
mechanism and to select a method for enzyme analysis. The conditions selected to measure the
activity of an enzyme would not be the same as those selected to measure the concentration of its
substrate. Several factors affect the rate at which enzymatic reactions proceed which are
temperature, pH, and concentration.

Substrate concentration
The maximum enzyme activity reached when all of enzyme combine with substrate. Thus, higher
substrate concentration the higher the rate of reaction (enzyme concentration is constant).

Figure 2 : Effect of substrate concentration

pH values
Enzyme are affected by pH values. High or low pH generally results in loss of enzyme activity. However,
the maximum enzyme activity will react at optimum pH values. Lower pH values mean higher

H+ concentrations and lower OH- concentrations.

Figure 3 : Effect of Temperature on Reaction Rate

REFERENCES

1. Dutta.R, Fundamental of Biochemical Engineering, Springer, New York, 2008, pg 31

2. Factors Affecting Enzyme Activity, 2012, Encyclopaedia Britannican , [Online]. [Accesses on
27th February 2018]. Available from World Wide Web:
http://www.britannica.com/EBchecked/topic/189245/enzyme/2123/Factors-affecting-
enzyme-activity
3. Bisswanger, H. Practical Enzymology, 2nd ed. Wiley- Blackwell, Weinheim, 2011, p299
4. Schomburg, D., Schomburg, I. (Eds.), Springer Handbook of Enzymes. Second Edition.
Springer, Berlin, 2009