Compartmented microfluidic bioreactor system using magnetic enzyme immobilisates for fast small‐scale biotransformation studies

Abstract

In the last decade, microfluidic bioreactor systems became increasingly important due to their high suitability for lab‐on‐a‐chip applications and resource‐saving experiments with small sample volumes. Here, a prototype of a microfluidic device for fast small‐scale investigations of enzymatic and biochemical reactions is introduced. Single or consecutive enzyme‐catalyzed reactions can be implemented within compartmented reaction environments separated by immiscible fluidic plugs. By immobilizing one of the reactants onto magnetic microcarriers, a fast and easy separation of the reaction products is possible allowing the realization of a sequence of different reaction steps with different enzymes and varying chemical environments. Besides permanent magnetic fields for separation processes, alternating electromagnetic fields can be applied to resuspend the carriers. This leads to an intense mixing as well as even microcarrier distribution within the compartment. In a proof of concept, kinetic studies of HRP immobilized onto polyvinyl alcohol‐magnetite composite microcarriers are presented. The results showed a specific enzyme activity of approximately 89 units per gram immobilized biocatalyst under the applied reaction conditions. In addition, results of recycling experiments point out the importance of the magnetically induced resuspension. While ten times reuse with immobilisate resuspension resulted in substrate conversion yields between 95 and 65%, the same experiment without the magnetically induced resuspension showed conversion yields below 10% over all cycles.

Automated Solid-Phase Protein Modification with Integrated Enzymatic Digest for Reaction Validation: Application of a Compartmented Microfluidic Reactor for Rapid Optimization and Analysis of Protein Biotinylation

“…In this study, we present an enhanced version of the reaction device presented in Hübner et al ( 2015 ). A schematic is given in Figure 3 .…”

“…Thus, the particles can sequentially enter reaction plugs filled with different reactants and react accordingly. In Hübner et al ( 2015 ), a precursor of the reactor was presented. In this study, we show an important enhancement of the reactor and its use for the screening of biotinylation conditions.…”

Microfluidic tools toward industrial biotechnology

“…A major contribution of microbioreactors to biological sciences is the improved knowledge of cellular responses in a dynamic and complex medium similar to in vivo or in situ systems. For instance, in bioprocesses, this technology can help understand the microbial responses caused by cellular interactions and environmental variations, evaluate microbial cell behavior, investigate kinetics of immobilized enzymes and biochemical reactions . These studies can contribute to improve processes knowledge since these effects represent a high impact on productivity …”

“…For instance, in bioprocesses, this technology can help understand the microbial responses caused by cellular interactions and environmental variations, 133 evaluate microbial cell behavior, 83 investigate kinetics of immobilized enzymes and biochemical reactions. 79,173,174 These studies can contribute to improve processes knowledge since these effects represent a high impact on productivity. 175 Despite the evident contribution of microbioreactors for bioprocess development, most of the applications with industrial focus have been to medical and pharmaceutical applied areas aiming to understand dynamic cells regulation 84,176 and production of new drugs.…”

Formulation of organic and inorganic hydrogel matrices for immobilization of β‐glucosidase in microfluidic platform

“…Inorganic silica‐based sol–gels can, for instance, be applied in microextractions, separation columns as well as immobilization of biomolecules . The latter case is of particular interest, as the entrapment of enzymes or other bioactive compounds, such as whole cells or immunoglobulins, in sol–gel matrices comes with advantages such as decreased consumption of the often expensive biocomponent as well as more controlled reaction conditions .…”