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T. Bayer, T. Wiesinger, S. Milker, F. Rudroff, M.D. Mihovilovic:
"Convergent Carbon Flux via 'Substrate Funneling' for the In Vivo Production of Reactive Aldehyde Species";
Vortrag: 8th European Meeting on OxiZymes, Wageningen; 03.07.2016 - 06.07.2016; in: "OxiZymes2016", (2016), S. 41.

The maximization of product titers of artificial enzyme cascades in vivo is highly desired. Strategies to enhance the carbon flux through de novo pathways such as the introduction of synthetic protein scaffolds and the knock-out of endogenous enzyme activities proved their applicability but were elaborate and time-consuming. Herein, an easy yet conceptually different approach called `substrate funneling´ was used for the optimization of an in vivo cascade for the preparation of reactive aldehyde species from primary aromatic alcohols and their subsequent aldol reaction with dihydroxyacetone (DHA) for the production of valuable polyhydroxylated compounds. Substrate funneling economically redirected the carbon flux from an undesired byproduct towards the target aldehyde intermediates 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 only 6 h reaction time.

flux optimization, synthetic enzyme cascade, substrate funneling, carboxylic acid reductase

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