Applicability of fluorescence-based sensors to the determination of kinetic parameters for O-2 in oxygenases

作者：Di Russo Natali V^*; Bruner Steven D; Roitberg Adrian E

来源：Analytical Biochemistry, 2015, 475: 53-55.

DOI：10.1016/j.ab.2015.01.009

• Oxygenases
• Enzyme assays
• Fluorescence quenching
• Oxygen
• Enzyme kinetics

Optical methods for O-2 determination based on dynamic fluorescence quenching have been applied to measure oxygen uptake rates in cell culture and to determine intracellular oxygen levels. Here we demonstrate the applicability of fluorescence-based probes in determining kinetic parameters for O-2 using as an example catalysis by a cofactor-independent oxygenase (DpgC). Fluorescence-based sensors provide a direct assessment of enzyme-catalyzed O-2 consumption using commercially available, low-cost instrumentation that is easily customizable and, thus, constitutes a convenient alternative to the widely used Clark-type electrode, especially in cases where chemical interference is expected to be problematic.

• 出版日期2015-4-15