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Forizymes-functionalised artificial forisomes as a platform for the production and immobilisation of single enzymes and multi-enzyme complexes

: Visser, F.; Müller, B.; Rose, J.; Prüfer, D.; Noll, G.A.

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Scientific Reports 6 (2016), Art. 30839, 15 S.
ISSN: 2045-2322
Zeitschriftenaufsatz, Elektronische Publikation
Fraunhofer IME ()

The immobilisation of enzymes plays an important role in many applications, including biosensors that require enzyme activity, stability and recyclability in order to function efficiently. Here we show that forisomes (plant-derived mechanoproteins) can be functionalised with enzymes by translational fusion, leading to the assembly of structures designated as forizymes. When forizymes are expressed in the yeast Saccharomyces cerevisiae, the enzymes are immobilised by the self-Assembly of forisome subunits to form well-structured protein bodies. We used glucose-6-phosphate dehydrogenase (G6PDH) and hexokinase 2 (HXK2) as model enzymes for the one-step production and purification of catalytically active forizymes. These structures retain the typical stimulus-response reaction of the forisome and the enzyme remains active even after multiple assay cycles, which we demonstrated using G6PDH forizymes as an example.