Liese, AndreasAndreasLieseHoltmann, DirkDirkHoltmann2026-08-242026-08-242025-03-31Technische Universität Hamburg (2025)https://hdl.handle.net/11420/64439In this study, scalable biocatalytic processes were developed in which unspecific peroxygenase (UPO) is used for selective oxyfunctionalization. The most important challenges in terms of enzyme production and stability, controlled electrochemical generation of cosubstrates, in situ product removal and semi-continuous operability were addressed. Robust UPO expression and downstream strategies enabled continuous operation, while detailed kinetic characterisation showed that a precise balance between substrate and hydrogen peroxide concentrations is required to prevent oxidative inactivation. Immobilised UPO such as His-tag, Twin-Strep-labelled and YSD (yeast surface display) variants, achieved high activity retention and operational stability in fluidised bed systems, enabling longer reaction cycles and simplified product recovery.enhttps://creativecommons.org/licenses/by/4.0/enzyme catalysisunspecific peroxygenase (UPO)biotechnologybubble columnTechnology::660: Chemistry; Chemical Engineering::660.6: BiotechnologyReport10.15480/882.18006