Galactose Oxidase‐Catalyzed Benchtop and Electrochemical Oxidation of C1‐Alkylated β‐D‐Galactopyranoside Derivatives

The reaction conditions of wild‐type galactose oxidase (GalOx)‐catalyzed oxidation of C‐1 alkylated β ‐D‐galactopyranoside derivatives are established and optimized. The benchtop oxidation reaction uses a three‐enzyme system comprising GalOx, catalase, and horseradish peroxidase (HRP). Under the optimized conditions, methyl‐ β ‐D‐galactopyranoside achieves 86% conversion after 24 h of reaction at room temperature. No previous study has investigated how the C‐1 position of β ‐D‐galactopyranoside derivatives influences GalOx–substrate interactions. To address this, a panel of C‐1 alkylated β ‐D‐galactopyranoside derivatives is synthesized and tested for the first time. The results show that although GalOx tolerates modifications at the C‐1 position, the size, shape, and electronic properties of the substituent significantly affect catalytic efficiency. The same β ‐D‐galactopyranoside derivatives are also investigated on an electrochemical setup, which replaces the HRP activator with electrochemical activation. The electrochemical activation method achieves comparable conversion and preserves substrate specificity, offering a promising and cost‐effective alternative to traditional enzymatic setups. This work provides a foundation for understanding C‐1 structural tolerance in GalOx–substrate interactions and demonstrates electrochemical activation as a promising and practical alternative to traditional enzymatic activation.