Journal ArticleDOI
Tailoring the S‐Selectivity of 2‐Succinyl‐5‐enolpyruvyl‐6‐hydroxy‐3‐cyclohexene‐1‐carboxylate Synthase (MenD) from Escherichia coli
Robert Westphal,Doris Hahn,Ursula Mackfeld,Simon Waltzer,Maryam Beigi,Michael Widmann,Constantin Vogel,Jürgen Pleiss,Michael Müller,Dörte Rother,Martina Pohl +10 more
TLDR
Opening the S‐pocket and simultaneous destabilization of the R‐pathway provides a potential general new strategy to enhance the S-selectivity of ThDP‐dependent enzymes.Abstract:
The thiamine diphosphate (ThDP)-dependent enzyme 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate synthase from Escherichia coli (EcMenD, E.C. 2.2.1.9) catalyzes the carboligation of α-ketoglutarate (α-KG) and various benzaldehyde derivatives with excellent chemo- as well as high R-selectivity (enantiomeric excess (ee) >93 %) to yield chiral α-hydroxy ketones. Based on the recently developed S-pocket concept, we engineered S-selective EcMenD variants by optimizing the steric properties and stabilization of the acceptor substrate in the S-pocket. Moreover, the moderate S-selectivity of the EcMenD variant I474A/F475G described recently for the carboligation of α-KG and benzaldehyde (ee=75 %) could be improved by selective destabilization of the R-pathway, which resulted in the variant I474A/F475G/R395Y (ee=85 % S). Subsequent investigation of the acceptor substrate range of this new variant revealed high S-selectivity especially with meta-substituted benzaldehydes, which gave access to 5-hydroxy-4-oxo-5-arylpentanoates with excellent enantioselectivities of up to 99 % ee S. Thus, opening the S-pocket and simultaneous destabilization of the R-pathway provides a potential general new strategy to enhance the S-selectivity of ThDP-dependent enzymes.read more
Citations
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Journal ArticleDOI
Building Bridges: Biocatalytic C-C-Bond Formation toward Multifunctional Products.
TL;DR: The review gives a selection of biocatalytic C–C-bond-forming reactions which have been investigated during the last 5 years and which have already been proven to be applicable for organic synthesis.
Journal ArticleDOI
The modular structure of ThDP‐dependent enzymes
Constantin Vogel,Jürgen Pleiss +1 more
TL;DR: An evolutionary pathway of ThDP‐dependent enzymes is proposed which explains the sequence and structure diversity of this family by three basic evolutionary events: domain recruitment, domain linkage, and structural rearrangement of catalytic domains.
Journal ArticleDOI
A tailor-made chimeric thiamine diphosphate dependent enzyme for the direct asymmetric synthesis of (S)-benzoins.
Robert Westphal,Constantin Vogel,Carlo Schmitz,Jürgen Pleiss,Michael Müller,Martina Pohl,Dörte Rother +6 more
TL;DR: This work has constructed a chimeric enzyme, which catalyzes the synthesis of various (S)-benzoins with excellent enantiomeric excess (>99%) and very good conversion.
Journal ArticleDOI
Regio- and Stereoselective Aliphatic-Aromatic Cross-Benzoin Reaction: Enzymatic Divergent Catalysis.
Maryam Beigi,Ekaterina Gauchenova,Lydia Walter,Simon Waltzer,Fabrizio Bonina,Thomas Stillger,Dörte Rother,Martina Pohl,Michael Müller +8 more
TL;DR: The regio- and stereochemistry of the product in the asymmetric aliphatic-aromatic cross-benzoin reaction can be controlled solely by choice of the appropriate enzyme or enzyme variant.
Journal ArticleDOI
Stereoselective Two-Step Biocatalysis in Organic Solvent: Toward All Stereoisomers of a 1,2-Diol at High Product Concentrations
TL;DR: This work combines cheap whole cell catalysts with a microaqueous solvent system to obtain 1-phenylpropane-1,2-diol (PPD), a versatile building block for pharmaceuticals, starting from inexpensive aldehyde substrates.
References
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