pH's Impact on Enzyme Activity

Introduction


A substance's PH value, which ranges from 0 to 14, can assist identify whether it is acidic or alkaline. Values above 7 are considered basic, while values below 7 are considered acidic. 7 nevertheless stands for impartial. A biological component called an enzyme is a protein that functions as a catalyst. They work as catalysts to quicken chemical reactions (Mutoh, Nakagawa, & Yamada, 1999). Catalase is one of the enzymes that helps hydrogen peroxide break down into water and oxygen molecules (Arabaci & Usluoglu, 2012). Catalase is a protein that guards against oxidative cell damage by interacting oxygen species. Enzymes are known to be pH specific, and therefore changes in pH value is likely to affect the catalytic activity of catalase (Eed, 2012). Catalase usually works best at low acidic level with pH values between 6.9 and 7.1. Therefore, the aim of this experiment is to determine how acidic condition or fluid affect the catalytic activity of the catalase.

Methods


The enzyme to be used in the study would be catalase obtained from the liver that has a higher amount of catalase. Hydrochloric acid (HCL) would be used to provide the acidic solution while hydrogen peroxide would be the substrate of the experiment. Sodium hydroxide would help to neutralize the intensity of the acidity of the HCL. The Hydrochloric acids of different concentrations are prepared to provide acidic levels of various pH values. Mixing HCL and different quantities of HCL produce difference pH values of acid. The resulting solutions are divided into five flasks depending on the acidity determined by the use of litmus paper. The solutions are then divided into five labeled flasks based on their acidity of the solutions as determined by litmus paper and pH value of 2, 4, 5, 6, and 7.

Liver of about 1 cm cube is chopped into pieces, grounded in some water and then little sand added to prompting the liver to produce catalase. The resulting mixture is filtered to remove the impurities. The filtrate obtained is then divided into 1cm cube tubes. 1 cm cube of hydrogen peroxide solution is then added to each tube. They are then closed tightly by use of cork. One of the tubes is inserted into a box filled with water and the initial level of water recorded to enable recording of any volume of oxygen gas produced as a result of a catalytic decomposition of hydrogen peroxide in the test tube into hydrogen and water molecules (Arabaci & Usluoglu, 2012). The oxygen produced appear as bubbles and increases the amount of water in the tube. The number of bubbles produced indicate the intensity of the enzymatic reaction. The amount of gas produced is derived by determining the difference between the final volume of water and the initial volume of water.

Results


pH values
Initial result
Final result


pH 7
4cm3
4.50 cm3


pH 6
4 cm3
4.25 cm3


pH 5
4 cm3
4.00 cm3


pH 4
4 cm3
4.00 cm3


pH 3
4 cm3
4.00 cm3


pH 2
4 cm3
4.00 cm3

The outcome of the experiment shows that the rate of bubble production was high in the tube with pH value 6 and 7. However, a negligible amount of bubbles occurred in those tubes with pH values 2 and 4. The amount of gas released in the experiment was measured with pH7 having the highest at 0.5 cm3 while pH 2, 3, 4 recording 0. The results are in the table below.

Discussion


A significant amount of gas release at pH 7 and the almost negligible amount of pH 2, 3, and 4 indicate that enzymes operate at specific pH values (Bartoszek & Sułkowski, 2006). The result also shows that enzyme catalase function well at low acidic conditions and in particular between pH 6 and 7. The increase in acidity from 6 toward two lead to affect the shape of catalase inhibiting its combination with hydrogen peroxide impairment of the catalase enzyme and therefore fails to carry out its catalytic function as evidenced by zero increase in the volume of water (Bartoszek & Sułkowski, 2006). The damage of the enzyme is due to competition between hydrogen ions and positive hydrogen atoms. In high acidic solutions, the concentration of the ions is greater, and this competes with a hydrogen atom to bind in electronegative atoms of the molecule causing impairment of the protein in catalase. Thus the experiment has supported the hypothesis that catalase is pH specific.


References


Arabaci, G., & Usluoglu, A. (2012). Catalytic Properties and Immobilization Studies of Catalase from Malva sylvestris L. Journal of Chemistry, 2013.


Bartoszek, M., & Sułkowski, W. W. (2006). The study of pH influence on bovine liver catalase using UV-VIS spectroscopy and spin labeling method. Polish Journal of Environment Studies, 41.


Eed, J. (2012). Factors Affecting Enzyme Activity. ESSAI, 10(1), 19.


Mutoh, N., Nakagawa, C. W., & Yamada, K. (1999). The role of catalase in hydrogen peroxide resistance in fission yeast Schizosaccharomyces pombe. Canadian journal of microbiology, 45(2), 125-129.

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