Caught in-between: System for in-flow inactivation of enzymes as an intermediary step in “plug-and-play” microfluidic platforms

The need for fast and comprehensive characterization of biocatalysts has pushed the development of new screening platforms based on microfluidics, capable of monitoring several parameters simultaneously, with new configurations of liquid handling, sample treatment and sensing. Modular microfluidics allows the integration of these newly developed approaches in a more flexible way towards increasing applicability of the microfluidic chips to different types of biocatalysts and reactions. A highly relevant operation in such a system is biocatalyst inactivation, which can enable the precise control of reaction time by avoiding the continuation of the reaction in another module or connecting tubes. Such control is important when different modules of reactors and/or sensing units are used and changed frequently. Here we describe the development, characterization and application of a module for rapid enzyme inactivation. The thermal inactivation platform developed is compared with a standard benchtop ThermoMixer in terms of inactivation efficiency for glucose oxidase and catalase. A higher activity loss was observed for enzyme inactivation under flow conditions (inactivation achieved at 120 s residence time at 338 K and 20 s residence time at 353 K) which indicated a high heat transfer to the fluid under dynamic conditions. Moreover, partial deactivation of the enzymes was observed for the continuous thermal inactivation module, when activity measurements were performed after 1 and 2 days following inactivation. The thermal inactivation unit presented can be easily integrated into modular microfluidic platforms and can be a useful addition for enzyme characterization and screening.