Substrate Walking for Selective Ene Reduction
Hugo Brasselet (TU Delft - Applied Sciences)
Jonathan Berger (TU Delft - Applied Sciences)
Elisa Arends (Student TU Delft)
Adriaan de Man (Student TU Delft)
Laura van der Weel (TU Delft - Applied Sciences)
Hessel van Dijk (ChiralVision)
Anke Hummel (Bielefeld University)
Harald Gröger (Bielefeld University)
Ulf Hanefeld (TU Delft - Applied Sciences)
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Abstract
Ene-reductases (EREDs) efficiently reduce activated C═C bonds, but their activity toward tetra-substituted alkenes has been largely unexplored. Here, we report the first systematic study demonstrating that several EREDs catalyze the stereoselective reduction of tri- and tetra-substituted cyclohexenones while investigating the influence of different cofactors. To enable a scalable application, the best enzyme (YqjM) was co-immobilized with glucose dehydrogenase on Ni–NTA cellulose beads, achieving >99% immobilization efficiency and high operational stability. The immobilized system was successfully scaled to 200 mL, reaching full substrate conversion without enzyme degradation over 9 days.
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