Asymmetric One‐Pot Total Synthesis of Tenuipesone A/B Through a Tunable Multienzyme Cascade

While biocatalysis has rapidly become a versatile tool in the synthetic chemist's toolbox, one of its main benefits, the broad cross‐compatibility of enzymes, which could lead to cell biochemistry‐like artificial metabolisms, is still underutilized in organic synthesis. Herein, we demonstrate the power of biocatalysis with an integrated single‐pot operation for the total synthesis of the fungal tenuipesone spirolactones directly from the biorefinery side stream furfural. An assembly of six enzymes at the core of a route design covering five linear steps delivers the tenuipesones in overall yields up to 72% (i.e. 94% per individual step). Careful selection of specific enzyme combinations allows to produce either of the enantiomers in excellent optical purities, facilitating the reassignment of the metabolites’ absolute configurations. This one‐pot process highlights the immense potential of complex multienzyme in vitro metabolisms and may inspire a much broader implementation of biocatalytic retrosynthesis to develop streamlined routes to other target scaffolds.