Evaluation of the Interactions of Some Alkali and Alkali Earth Metallic Chlorides on Guaiacol Oxidation by Peroxidase from Watermelon Rind

Abstract

Aim: This study evaluated Interactions of some alkali and alkali earth metallic chlorides on guaiacol oxidation by peroxidase from watermelon rind. Study Design:  In vitro enzyme assay. Place and Duration of Study: Department of Biochemistry, Faculty of Life Sciences, Ambrose Alli University, Ekpoma, Edo State, Nigeria between September  2023 to November  2023. Methodology: The kinetics of the oxidation of Guaiacol by the crude peroxidase from the rind of watermelon in the presence of varying concentrations of the chloride salts of Na, K, Mg and Ca was determined spectrophotometrically by monitoring the oxidation of Guaiacol to produce a brown tetraguaiacol monitored at a wavelength of 470nm.  The various salt concentrations were varied between 0.5 mM and 3 mM.  Each of the reaction mixtures used in the kinetic study comprised;  2.3 mL of 0.6 M sodium acetate buffer (pH 5.4), 0.2 mL of 0.02 mM Guaiacol, 0.1 mL of crude extract, 0.2 mL of varying concentration chloride salt, 0.2 mL of 2 mM of H2O2 added last to start the reaction. The final concentration of H2O2 in the 3 mL assay was 0.13 mM. The total volume of the reaction mixture was 3 mL. The absorbance was read every 2 seconds for one minute after adding hydrogen peroxide using a stop-clock. The control had no metal ion but replaced with distilled water Results: Results showed that within a salt concentration range of 0.5mM to 3Mm, chlorides of  Na, K and Mg  activated the peroxidase from the rind of watermelon while CaCl2 reduced the activity in a concentration dependent manner. The optimal salt concentration of these cation activators was 0.5 mM. Conclusion: These findings are of great importance to industries in understanding the mechanism of action of peroxidase from the rind of watermelon, especially as the search for cheap and alternative sources of peroxidases continues.