Substrate Mass Conservation in Enzyme Catalyzed Amylolytic Activity

Abstract

Aims: The aims of the research were 1), to derive simple equations that can be used to determine mass concentration of reaction mixture components, and 2), to determine the mass concentration of free substrate, total mass concentration of substrate involved in enzyme-substrate complex formation, and mass concentration of partially digested parent starch, otherwise called fragments at the end of different durations of assay. Study Design: Theoretical and Experimental. Place and Duration of Study: Department of Chemistry and Biochemistry, Research Division, Ude International Concepts LTD (862217), B. B. Agbor, Delta State, Nigeria; Owa Alizomor Secondary School, Owa Alizomor, Ika North East, Delta State, Nigeria. The research, including derivation of equations lasted between 18th April, 2017 and 24th June, 2017. Methodology: Bernfeld method of enzyme assay was used. Assays were carried out on Aspergillus oryzea salivary alpha amylase. Various parameters were determined by substituting relevant experimental data to the formulated equations. Results: The mass concentrations of the free undigested starch, the enzyme-substrate complex, and the fragments showed decreasing trend with time. The exception was at highest duration with respect to the concentration of the fragment. The concentrations of the remaining substrate calculated using two different approaches, and the new approach (Eq. (31)) in this research were not statistically different (P > 0.05). The difference between the change in the concentration of the substrate per unit time, D[S0]/min, and the corresponding product formed per unit time, [P]/min, was not statistically different (P > 0.05). Conclusion: Different algebraic equations were successfully derived and were used to determine various components of the hydrolyzate at the end of the duration of assay. The hydrolyzate is composed of different components. The sum of the components was very similar to total concentration of substrate, in conformity with substrate mass conservation law.