Nucleophile Promiscuity of Engineered Class II Pyruvate Aldolase YfaU from E. Coli |
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Authors: | Dr. Karel Hernández Dr. Jesús Joglar Dr. Jordi Bujons Dr. Teodor Parella Prof. Dr. Pere Clapés |
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Affiliation: | 1. Chemical Biology and Molecular Modelling, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Barcelona, Spain;2. Servei de Ressonància Magnètica Nuclear, Universitat Autònoma de Barcelona, Bellaterra, Spain |
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Abstract: | Pyruvate‐dependent aldolases exhibit a stringent selectivity for pyruvate, limiting application of their synthetic potential, which is a drawback shared with other existing aldolases. Structure‐guided rational protein engineering rendered a 2‐keto‐3‐deoxy‐l ‐rhamnonate aldolase variant, fused with a maltose‐binding protein (MBP‐YfaU W23V/L216A), capable of efficiently converting larger pyruvate analogues, for example, those with linear and branched aliphatic chains, in aldol addition reactions. Combination of these nucleophiles with N‐Cbz‐alaninal (Cbz=benzyloxycarbonyl) and N‐Cbz‐prolinal electrophiles gave access to chiral building blocks, for example, derivatives of (2S,3S,4R)‐4‐amino‐3‐hydroxy‐2‐methylpentanoic acid (68 %, d.r. 90:10) and the enantiomer of dolaproine (33 %, d.r. 94:6) as well as a collection of unprecedented α‐amino acid derivatives of the proline and pyrrolizidine type. Conversions varied between 6–93 % and diastereomeric ratios from 50:50 to 95:5 depending on the nucleophilic and electrophilic components. |
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Keywords: | aldol reactions amino acids biocatalysis pyruvate aldolases |
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