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Online Measurement of Oxygen‐Dependent Enzyme Reaction Kinetics
Author(s) -
Meissner Murray P.,
Nordblad Mathias,
Woodley John M.
Publication year - 2018
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700577
Subject(s) - chemistry , kinetics , catalysis , oxygen , chemical kinetics , reaction rate , titration , mass transfer , redox , reaction rate constant , chemical reaction engineering , glucose oxidase , chromatography , organic chemistry , enzyme , physics , quantum mechanics
As the application of biocatalysis to complement conventional chemical and catalytic approaches continues to expand, an increasing number of reactions involve poorly water‐soluble substrates. At required industrial concentrations necessary for industrial implementation, this frequently leads to heterogeneous reaction mixtures composed of multiple phases. Such systems are challenging to sample, and therefore, it is problematic to measure representative component concentrations. Herein, an online method for following the progress of oxygen‐dependent reactions through accurate measurement of the oxygen mass balance in the gas phase of a reactor is demonstrated and validated. The method was successfully validated and demonstrated by using two model reactions: firstly, the oxidation of glucose by glucose oxidase and, secondly, the Baeyer–Villiger oxidation of macrocyclic ketones to lactones. Initial reaction rate constants and time‐course progressions calculated from the oxygen mass balance were validated against conventional online methods of dissolved oxygen tension and pH titration measurements. A feasible operating window and the sensitivity to dynamic changes of reaction rates were established by controlling oxygen transfer through the operating parameters of the reactor. Such kinetic data forms the basis for reaction characterisation, from which bottlenecks may be made evident and directed improvement strategies can be identified and implemented.