A Screening Platform to Identify and Tailor Biocompatible Small‐Molecule Catalysts

Abstract

Interfacing biocompatible, small‐molecule catalysis with cellular metabolism promises a straightforward introduction of new function into organisms without the need for genetic manipulation. However, identifying and optimizing synthetic catalysts that perform new‐to‐nature transformations under conditions that support life is a cumbersome task. To enable the rapid discovery and fine‐tuning of biocompatible catalysts, we describe a 96‐well screening platform that couples the activity of synthetic catalysts to yield non‐canonical amino acids from appropriate precursors with the subsequent incorporation of these nonstandard building blocks into GFP (quantifiable readout). Critically, this strategy does not only provide a common readout (fluorescence) for different reaction/catalyst combinations, but also informs on the organism's fitness, as stop codon suppression relies on all steps of the central dogma of molecular biology. To showcase our approach, we have applied it to the evaluation and optimization of transition‐metal‐catalyzed deprotection reactions.

Hydrodechlorination of Aryl Chlorides Under Biocompatible Conditions

“… 29 Work is also being conducted to streamline the process of screening for biocompatible catalysts. 30 These works show the feasibility and application of modifying organic synthetic methods to be compatible with microbial growth.…”

Tools and Methods for Investigating Synthetic Metal-Catalyzed Reactions in Living Cells

“…For example, Waymouth, Wender, and co-workers developed a luciferase reporter system to screen for complexes capable of facilitating allyl carbamate cleavage . Mayer and co-workers created a 96-well plate screening platform that measures both catalyst activity and organism fitness …”

“…75 Mayer and coworkers created a 96-well plate screening platform that measures both catalyst activity and organism fitness. 134…”

Interfacing non-enzymatic catalysis with living microorganisms

“…60 Finally, it is also noteworthy that a 96-well plate assay has been developed to rapidly identify biocompatible transition metal catalysts. 61 This system couples biocompatible catalyst activity with a fluorescence readout via incorporation of an in situ synthesized non-canonical amino acid into green fluorescent protein through Amber stop-codon suppression, a strategy that has also been used to 'addict' E. coli to abiotic reactions. 62 This methodology was hypothesised to be widely applicable to any chemical transformation that yields a noncanonical amino acid and therefore presents a general platform for the identification of new biocompatible reactions in the future.…”