Abstract

The aim of this study is to find optima of operating conditions of phosphorylation enzymatic reaction of glucose within differential pH sensors device. Five variables were studied, i.e., amount of enzyme (7.45-44.7 µl), buffer concentration (20-50mM), pH of buffer (6.8 to 8.1), ATP concentration (0.2-2mM) and Mg⁺² concentration (1.2 to 6mM).

The kinetic study indicated optima of amount of enzyme of (30µl), buffer concentration of (40mM), pH value of buffer of (7.6), ATP concentration of (1.2mM) and Mg²⁺ concentration of (2.2mM) for phosphorylation of 1g/l glucose concentration sample. A calibration curve of glucose quantification was done for low glucose concentration range i.e. from 0 to 1 g/l. This low range makes the assay of this study efficient to be used in many applications. For instance, the assay can be used in glucose quantification during cultivation of variety kinds of cells at low glucose concentration. The assay was developed by using HEPES buffer as new carrying buffer system.

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