[Zurück]

T. Bayer, S. Milker, T. Wiesinger, M.D. Mihovilovic, F. Rudroff:
"'Substrate/redox funneling' as a novel flux optimization tool for synthetic enzyme cascades in vivo";
Poster: 5th International Conference on Novel Enzymes, Groeningen; 11.10.2016 - 14.10.2016; in: "5th International Conference on Novel Enzymes", (2016).

The maximization of product titers of artificial enzyme cascades in vivo is highly desired.1 Strategies to enhance the carbon flux through de novo pathways such as the introduction of synthetic protein scaffolds2 and the knock-out of endogenous enzyme activities3 proved their applicability beyond doubt but were elaborate and time-consuming. Herein, an easy to apply yet conceptually different approach called `substrate/redox funneling´ was used for the optimization of an in vivo cascade for the preparation of reactive aldehyde intermediates from primary (aromatic) alcohols and their subsequent aldol reaction with dihydroxyacetone (DHA) for the production of valuable polyhydroxylated compounds. `Substrate/redox funneling´ economically redirected the carbon flux from undesired carboxylate byproducts toward the target aldehydes solely by the introduction of an enzyme with opposing functional group activity. The potential of the optimized synthetic pathway, combined with a simple solid-phase extraction purification protocol, was demonstrated by the production of the aldol (3S,4R)-1,3,4-trihydroxy-5-phenylpentan-2-one from 2-phenyl ethanol and DHA on preparative scale yielding 70% pure aldol in short reaction times.
Since `substrate/redox funneling´ is fully compatible with other cascade type reactions and can be applied to reverse other byproduct forming reactions, it offers a novel and complementary flux optimization tool for synthetic enzyme cascades in vivo.

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