The effect of buffer pH on enzyme activity

Abstract

This study investigates the effect of pH on enzyme-catalyzed activity, with an emphasis on the involvement of wheat germ acid phosphatase in dephosphorylation processes. Temperature, pH, and enzyme concentration all have a substantial impact on enzyme activity and substrate interaction. Wheat germ acid phosphatase was chosen for this work. The activity of the enzyme in dephosphorylating p-nitrophenyl phosphate (p-NPP) into p-nitrophenol (p-NP) was evaluated using a stopped assay approach. The reaction was stopped by introducing sodium hydroxide, which ionises p-NP and produces a yellow anion visible at 405 nm. The experiment measured product production in nmol per minute and included three sections: Created a standard for the amount of p-NP with its absorbance at 405 nm; Found the ideal incubation period for maximal product generation by wheat germ acid phosphatase; Evaluated the effect of pH on enzyme activity measuring the first reaction velocity in various pH settings. The results showed that when the buffer pH diverged from the ideal value of 5.5, catalysis rates reduced dramatically, implying that high pH levels may modify the protein structure of the enzyme and reduce its catalytic effectiveness. This work emphasises the importance of pH in enzymatic activity and its potential impacts on enzyme structural integrity.