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- W4284689812 abstract "A key bottleneck in the microbial production of therapeutic plant metabolites is identifying enzymes that can improve yield. The facile identification of genetically encoded biosensors can overcome this limitation and become part of a general method for engineering scaled production. We have developed a combined screening and selection approach that quickly refines the affinities and specificities of generalist transcription factors; using RamR as a starting point, we evolve highly specific (>100-fold preference) and sensitive (half-maximum effective concentration (EC50) < 30 μM) biosensors for the alkaloids tetrahydropapaverine, papaverine, glaucine, rotundine and noscapine. High-resolution structures reveal multiple evolutionary avenues for the malleable effector-binding site and the creation of new pockets for different chemical moieties. These sensors further enabled the evolution of a streamlined pathway for tetrahydropapaverine, a precursor to four modern pharmaceuticals, collapsing multiple methylation steps into a single evolved enzyme. Our methods for evolving biosensors enable the rapid engineering of pathways for therapeutic alkaloids. A combined screening and selection approach enables the evolution of the generalist transcription factor RamR into specific and sensitive biosensors for various alkaloids and in turn a streamlined pathway for tetrahydropapaverine biosynthesis." @default.
- W4284689812 created "2022-07-08" @default.
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- W4284689812 date "2022-07-07" @default.
- W4284689812 modified "2023-10-17" @default.
- W4284689812 title "Using fungible biosensors to evolve improved alkaloid biosyntheses" @default.
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- W4284689812 doi "https://doi.org/10.1038/s41589-022-01072-w" @default.
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