New combined kinetic and thermodynamic approach to model glucose-6-phosphate dehydrogenase activity and stability

Process Integration for the Disruption of Candida guilliermondii Cultivated in Rice Straw Hydrolysate and Recovery of Glucose-6-Phosphate Dehydrogenase by Aqueous Two-Phase Systems

“…Ulusu et al [34] studied the purification of glucose-6-phosphate dehydrogenase from bovine lens by affinity chromatography and observed that the purified enzyme had K M of 0.008 and 0.035 mM for NADP + and G6P, respectively. Hasmann et al [5] showed that Michaelis constant for G6PD of S. cerevisiae was almost independent of the presence of cell debris (K M =49.3-49.4 μM), although these results were not in accordance with ours. Nevertheless, the present data are in good agreement with those of G6PD from Aspergillus aculeatus (75 μM) [36].…”

“…The development of a purification procedure requires the variation of factors to get the optimum intracellular protein recovery, namely polymer concentration, polymer molar mass, pH, and temperature system. Other authors analyzed stability of G6PD, and their results allowed setting the parameters temperature and pH system in 25°C and 7.5, respectively [5,24]. Therefore, we used a 2 2 central composite design to identify which ATPS composition (independent variables such as M PEG and TLL) is able to maximize G6PD purification factor and yield in the bottom phase (PF B , Y B ) of integrated process (Table 2).…”

“…The properties of oxido-reductase enzymes considerably increased their importance and application. Owing to its high catalytic activity and substrate specificity, G6PD has been extensively used as reagent in varied enzymatic assays aiming determinations of ATP and hexose concentrations [5]. Besides, using G6PD in enzyme-based biosensor would be a good option due to advantages as specific measurement of target analyte in complex matrices (e.g., blood, food products, and environmental samples) [6,7].…”

Liquid–liquid extraction of biomolecules: an overview and update of the main techniques

“…The results suggested that the concentrated enzyme solution originated from the extraction process was slightly more effective as a biocatalyst than the initial cell-free extract. 140 The versatility of reversed micellar systems have been confirmed and leads to potential further applications in several areas besides purification. These aggregates have been depicted as passive nano-reactors that can be used as organic media to perform reactions such as: hydrolysis of triglycerides, esterification of palmitic oil, hydrolysis of olive oil, ibuprofen synthesis, etc.…”

A new enzymatic process for the treatment of phenolic pollutants

“…Such a bell-shaped behavior of enzyme activity vs. temperature was already observed for most enzymatic systems, the position of the optimum activity being directly related to the temperature of maximum stability of the enzyme tertiary structure in water. For example, similar trends were observed for mycelium-bound carboxylesterases (Converti et al 2002), lipases (Pastorino et al 2004), dehydrogenases (Hasmann et al 2007), proteases (Viana et al 2010) and oxidases (Porto et al 2006). The effect of temperature on the tyrosinase activity was confirmed by the behavior of the oxidation efficiency that, as expected, exhibited a minimum value (78.6%) at T = 15°C, increased with temperature up to a maximum (88.1-88.5%) in the range 25 ≤ T ≤ 45°C and then decreased to only 30.9% at 55°C.…”