Constructing Biocatalytic Cascades: In Vitro and in Vivo Approaches to de Novo Multi-Enzyme Pathways

TLDR

Sequential biocatalytic cascades enable the synthesis of complex chemicals from simple precursors, and as the biocatalyst toolbox expands, new retrosynthetic routes emerge, but implementing such cascades requires careful consideration of whether they are constructed in vitro or in vivo. This Perspective outlines the relative merits of in vitro, in vivo, and hybrid approaches for building biocatalytic cascades and highlights recent developments. The authors discuss factors influencing the design and implementation of purely enzymatic or chemoenzymatic, one‑pot, multistep pathways.

Abstract

The combination of sequential biocatalytic reactions, via non-natural synthetic cascades, is a rapidly developing field and leads to the generation of complex valuable chemicals from simple precursors. As the toolbox of available biocatalysts continues to expand, so do the options for biocatalytic retrosynthesis of a target molecule, leading to alternative routes employing enzymatic transformations. The implementation of such cascade reactions requires careful consideration, particularly with respect to whether the pathway is constructed in vitro or in vivo. In this Perspective, we describe the relative merits of in vitro, in vivo, and hybrid approaches to building biocatalytic cascades and showcase recent developments in the area. We also highlight the factors that influence the design and implementation of purely enzymatic or chemoenzymatic, one-pot, multistep pathways.