A H₂O₂ generation-detection-regulation integrated platform for boosting the efficiency of peroxygenase-catalysed C–H oxidative hydroxylation

The peroxygenases are ideal biocatalysts for the selective oxyfunctionalisation of stable C–H bonds. However, the catalytic efficiency of this approach is limited due to enzyme lability toward oxidant H₂O₂. Although the reported in-situ H₂O₂ generation system enables the stable biocatalytic process without deactivating the enzyme, the greatest catalytic potential of peroxygenases still cannot be fulfilled effectively. To address the above issue, a H₂O₂ generation-detection-regulation platform that integrated an effective organocatalyst-driven H₂O₂ generation system, a precise electrochemical H₂O₂ real-time detection device, and a convenient H₂O₂ regulation strategy was first developed. The suitable range of H₂O₂ generation rate for maximizing the catalytic efficiency of peroxygenases while minimizing inactivation of the enzyme was firstly obtained by simply adjusting the amount of organocatalyst. According to the determined suitable range, the C–H oxyfunctionalisation efficiency of peroxygenases for each substrate was significantly boosted, achieving ∼3-fold of the reported highest turnover frequency.