Fully exploiting the potential of enzymes in cell‐free biocatalysis requires stabilization of the catalytically active proteins and their integration into efficient reactor systems. Although in recent years initial steps towards the immobilization of such biomolecules in metal–organic frameworks (MOFs) have been taken, these demonstrations have been limited to batch experiments and to aqueous conditions. Here we demonstrate a MOF‐based continuous flow enzyme reactor system, with high productivity and stability, which is also suitable for organic solvents. Under aqueous conditions, the stability of the enzyme was increased 30‐fold, and the space–time yield exceeded that obtained with other enzyme immobilization strategies by an order of magnitude. Importantly, the infiltration of the proteins into the MOF did not require additional functionalization, thus allowing for time‐ and cost‐efficient fabrication of the biocatalysts using label‐free enzymes.
Um das Potenzial von Enzymen in der zellfreien Biokatalyse voll auszuschöpfen, ist eine Stabilisierung der katalytisch aktiven Proteine und deren Integration in effiziente Reaktorsysteme erforderlich. Obwohl in den letzten Jahren erste Schritte zur Immobilisierung solcher Biomoleküle in metallorganischen Gerüsten (MOFs) unternommen wurden, waren diese Demonstrationen auf Batch-Experimente und wässrige Bedingungen beschränkt. Hier demonstrieren wir ein kontinuierliches Enzymreaktorsystem auf MOF-Basis mit hoher Produktivität und Stabilität, das auch für organische Lösungsmittel geeignet ist. Unter wässrigen Bedingungen wurde die Stabilität des Enzyms um das 30-fache erhöht, und die Raum-Zeit-Ausbeute übertraf die mit anderen Enzym-Immobilisierungsstrategien erzielten Werte um eine Größenordnung. Es ist herauszuheben, dass die hier gezeigte Infiltration der Proteine in das MOF keine zusätzliche Funktionalisierung erfordert, was eine zeit-und kosteneffiziente Herstellung der Biokatalysatoren mit markierungsfreien Enzymen ermöglicht.
The application potential of enzymes is undoubtedly very high. However, despite the very large number of different enzymes and enzyme activities, the number of industrial enzyme processes is comparatively small. The particular challenge often lies in transferring promising laboratory processes to an industrial scale. Here, the required performance parameters, such as enzyme stability or productivity, must be achieved. On the one hand, this can be achieved by improving the enzymes. On the other hand, the key performance indicators can often only be achieved by using technical systems in the sense of process intensification. In enzymatic processes, immobilization of enzymes is often the means of choice to enable technical processes. The aim of this article is to outline the most important enzyme immobilization methods and to summarize the most important performance indicators of immobilized enzymes. Finally, the different immobilization methods and performance indicators are compared in a case study with unspecific peroxygenase.
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