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Enzyme kinetics assay in ionic liquid-based reaction media by means of Raman spectroscopy and multivariate curve resolution |
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| Authors: | Mercedes López-Pastor Ana Domínguez-Vidal María José Ayora-Cañada Miguel Valcárcel |
| Institution: | a Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164AC, A-1060 Wien, Austria b Department of Physical and Analytical Chemistry, University of Jaen, Paraje de las Lagunillas s/n, E-23071, Jaén, Spain c Department of Analytical Chemistry, University of Cordoba, Campus of Rabanales, Marie Curie Annex Building, E-14071 Córdoba, Spain |
| Abstract: | Enzymatic hydrolysis of p-nitrophenylphosphate by alkaline phosphatase in binary mixtures of water and 1-ethyl-3-methylimidazolium tetrafluoroborate (emimBF4) was monitored with Raman microspectroscopy. Concentrations of emimBF4 in the studied ionic liquid/water solvent systems ranged from 0 to 75% v/v. Multivariate curve resolution-alternating least squares (MCR-ALS) was successfully applied to the recorded Raman spectra in order to retrieve the concentration profiles and pure Raman spectra of the different species involved in the reaction. Michaelis-Menten constant (KM) and maximum rate (Vmax) of the reaction were calculated from the initial reaction phase for the different solvent systems studied, to investigate the effect of increasing concentration of the ionic liquid on the kinetic behavior. From this study, it was found that the ionic liquid inhibits the reaction under study decreasing both Vmax and KM. |
| Keywords: | Raman Ionic liquids Enzymatic kinetics MCR-ALS ALP |
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