Kinetic Analysis Of P-Nitrophene Reaction Lab

INTRODUCTION
Enzyme is a biological catalyst that acts on a molecule called substrate and it also significantly speed up a chemical reaction by lowering the activation energy. In order to learn about the enzyme and its behaviour, this lab practical is conducted to examine the kinetic of the enzyme alkaline phosphatase. As illustration, when alkaline phosphatase is added to a substrate called p-Nitrophenyl phosphate (colourless in alkaline solution), a series of reaction takes place and eventually releases a product called p-nitrophenol (yellow in alkaline pH) by enzymatic hydrolysis. As more product formed, the solution turns yellow and this change in colour can be monitored by the spectrophotometer. This can be done by observing the colour changes over 20 minutes and record the value obtained as Vo (absorbance value). Moreover, a second experiment will be performed but this time it will be with the addition of the
The result shows in Table 1 indicate that as the substrate concentration increases, the absorbance value will also increase. This is because an increase in substrate concentration allow more substrates to bind with the available enzyme’s active sites and results in an increase of products formed which led to high V0 (Lim, 2016). Various compounds can reduce the enzyme activity by directly or indirectly interfere with the functioning of the active site of an enzyme and thus, reducing the activity. This can be done by adding inhibitor to an enzyme catalyse reaction which can caused the reaction to slow down and releases less product compares to a reaction without inhibitor. For example, when a substrate concentration of 0.04 mM is added to a solution without inhibitor, it will produce a larger V0 (0.39AU) compares to a solution with inhibitor (0.18AU) (Table