Flexible hydroxylation of small molecules through single step biocatalysis

Overview

Flexible hydroxylation of small molecules through single step biocatalysis leverages Unspecific Peroxygenases (UPOs) to catalyze the insertion of oxygen into non-activated C-H bonds of aromatic and aliphatic substrates. This method simplifies industrial bio-transformation by using inexpensive chemical inputs and operating under mild aqueous conditions. UPOs offer advantages over traditional oxygenase enzymes, providing regio- and stereo-selective hydroxylation with high efficiency, making them ideal for sustainable chemical synthesis.

Technical specifications

• UPOs Characteristics: Operate with high efficiency in mild aqueous conditions, making them cost-effective and environmentally friendly.
• Expression System: UPOs are expressed in a P. pastoris production strain, ensuring robust expression and secretion.
• Screening Campaign: A diverse library of UPOs is screened against various small molecule substrates, optimizing reaction conditions for expression, stability, and kinetics.
• Reaction Optimization: Includes pH profiling, H2O2 delivery, temperature adjustments, and testing of co-solvents/detergents to enhance mass transfer.
• Production Process: Successful UPOs are scaled for high-volume production, with processes for biomass removal and purification via filtering and organic solvent extraction.

Technology readiness level

This technology is at TRL 5, indicating that it has been validated in a relevant environment, with UPOs ready for advancement to gram-scale production and further optimization for commercial applications.