2024 AIChE Annual Meeting
Directed Evolution of Oxidized Bicyclic Monoterpene Biosensors
Terpenes are the largest and most diverse class of naturally occurring molecules with various biological activities, making them valuable for pharmaceuticals and industrial products. Despite their diverse applications, sourcing terpenes is challenging because most are extracted from their native producers, and several are minor products of chemical reactions. The oxidized bicyclic monoterpenes myrtenol and verbenol are extensively used as fragrances and flavorings and possess several medicinal properties, making them promising candidates for therapeutic agents and the treatment of certain diseases. However, their natural abundance is less than 0.70% and constitutes less than 40% of the native oxidation products of α-pinene. The optimization of the myrtenol and verbenol biosynthetic pathways may be achieved via a directed evolution approach. However, this approach accumulates a large number of enzyme variants that are difficult to efficiently screen using existing technology. We propose using transcription factor-based genetically encoded biosensors for this purpose. By engineering transcription factor proteins through multiple rounds of directed evolution, biosensors can be tailored for a specific molecule. Here, we report the initial screening and tailoring of different biosensor constructs with verbenol and myrtenol.