1. Extended substrate range of thiamine diphosphate-dependent MenD enzyme by coupling of two C-C-bonding reactions
- Author
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Sascha Ferlaino, Alexander Fries, Simon Waltzer, Shiromi Baier, Matthias Schapfl, Michael Müller, and Georg A. Sprenger
- Subjects
1,2-ADDITIONS ,Cyclohexanecarboxylic Acids ,Stereochemistry ,Decarboxylation ,CARBOLIGATIONS ,INGENIERÍAS Y TECNOLOGÍAS ,010402 general chemistry ,01 natural sciences ,Applied Microbiology and Biotechnology ,Biotecnología Industrial ,Substrate Specificity ,THDP-DEPENDENT ENZYME ,chemistry.chemical_compound ,Cyclohexenes ,ALDOLASE EDA ,Glyoxylic acid ,MEND ,biology ,010405 organic chemistry ,Chemistry ,Aldolase A ,Enantioselective synthesis ,Regioselectivity ,General Medicine ,Bioprocesamiento Tecnológico, Biocatálisis, Fermentación ,Keto Acids ,0104 chemical sciences ,4-HYDROXY-2-OXOGLUTARATE ,Biocatalysis ,Electrophile ,biology.protein ,Michael reaction ,STETTER-LIKE 1,4-ADDITIONS ,Thiamine Pyrophosphate ,Biotechnology - Abstract
Carboligations catalyzed by aldolases or thiamine diphosphate (ThDP)-dependent enzymes are well-known in biocatalysis to deliver enantioselective chain elongation reactions. A pyruvate-dependent aldolase (2-oxo-3-deoxy-6-phosphogluconate aldolase [EDA]) introduces a chiral center when reacting with the electrophile, glyoxylic acid, delivering the (S)-enantiomer of (4S)-4-hydroxy-2-oxoglutarate [(S)-HOG]. The ThDP-dependent enzyme MenD (2-succinyl-5-enol-pyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate synthase (SEPHCHC synthase)) enables access to highly functionalized substances by forming intermolecular C?C bonds with Michael acceptor compounds by a Stetter-like 1,4- or a benzoin-condensation 1,2-addition of activated succinyl semialdehyde (ThDP adduct formed by decarboxylation of 2-oxoglutarate). MenD-catalyzed reactions are characterized by high chemo- and regioselectivity. Here, we report (S)-HOG, in situ formed by EDA, to serve as new donor substrate for MenD in 1,4-addition reactions with 2,3-trans-CHD (2,3-trans-dihydroxy-cyclohexadiene carboxylate) and acrylic acid. Likewise, (S)-HOG serves as donor in 1,2-additions with aromatic (benzaldehyde) and aliphatic (hexanal) aldehydes. This enzyme cascade of two subsequent C?C bond formations (EDA aldolase and a ThDP-dependent carboligase, MenD) generates two new stereocenters. Fil: Schapfl, Matthias. Universität Stuttgart; Alemania Fil: Baier, Shiromi. Universität Stuttgart; Alemania Fil: Fries, Alexander Erich. Albert Ludwigs University of Freiburg; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ferlaino, Sascha. Albert Ludwigs University of Freiburg; Alemania Fil: Waltzer, Simon. Albert Ludwigs University of Freiburg; Alemania Fil: Müller, Michael. Albert Ludwigs University of Freiburg; Alemania Fil: Sprenger, Georg A.. Universität Stuttgart; Alemania
- Published
- 2018