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- W2979237553 abstract "ABSTRACT Insertions and deletions (InDels) are frequently observed in natural protein evolution, yet their potential remains untapped in laboratory evolution. Here we introduce a transposon mutagenesis approach (TRIAD) to generate libraries of random variants with short in-frame InDels, and screen TRIAD libraries to evolve a promiscuous arylesterase activity in a phosphotriesterase. The evolution exhibits features that are distinct from previous point mutagenesis campaigns: while the average activity of TRIAD variants is more deleterious, a larger proportion has successfully adapted for the new activity, exhibiting different functional profiles: ( i ) both strong and weak trade-off in original vs promiscuous activity are observed; ( ii ) trade-off is more severe (10- to 20-fold increased k cat / K M in arylesterase with ∼100-fold decreases in the original phosphotriesterase activity) and ( iii ) improvements show up in k cat rather than K M , suggesting novel adaptive solution. These distinct features make TRIAD an alternative to widely used point mutagenesis, providing access to functional innovations and traversing unexplored fitness landscape regions." @default.
- W2979237553 created "2019-10-10" @default.
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- W2979237553 date "2019-10-02" @default.
- W2979237553 modified "2023-09-27" @default.
- W2979237553 title "Access to unexplored regions of sequence space in directed enzyme evolutionviainsertion/deletion mutagenesis" @default.
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- W2979237553 doi "https://doi.org/10.1101/790014" @default.
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