Non-native Metabolon-based Bioelectrocatalytic System Using Chimeric Invertase-Glucose Dehydrogenase

Abstract

Enzymes are well recognized as very potent biocatalysts in the natural world, exerting a substantial influence on several metabolic processes. Enzymes often exist in a state that might potentially exhibit increased reaction efficiency inside the biological system via the formation of a multienzyme complex. In the present work, an in vitro system was developed by constructing a non-native metabolon by the deliberate conjugation of two enzymes originating from distinct sources. In this study, we have discovered the key parameters that have the potential to significantly impact the efficiency of the reaction. Glucose dehydrogenase (GDHαγ) was chosen as a representative enzyme due to its established credibility and extensively verified characteristics in prior studies. The coupling enzyme used for this study was invertase (INV), which has the ability to create glucose as an intermediate. In order to merge the two enzymes, a peptide spacer was used as a connecting element between GDHαγ and INV. The catalytic activity was significantly impacted by the expression order of GDHαγ and INV, and the catalytic activity exhibited notable variations based on the spacer sequence. The criticality of regulating the distance and orientation between the coupling enzymes in a cascade reaction has been identified.