Acyloin formation by benzoylformate decarboxylase from Pseudomonas putida

Abstract

Whole cells and cell extracts of Pseudomonas putida grown in a medium containing ammonium mandelate have the capacity to produce the acyloin compound 2-hydroxypropiophenone when incubated with benzoylformate and acetaldehyde. Benzaldehyde and benzyl alcohol were formed as reaction by-products. The enantiomeric excess of the 2-hydroxypropiophenone product was found to be 91 to 92%. The absolute configuration of the enzymatically prepared product at the carbinol carbon was found to be S. The thiamine PP1-linked enzyme benzoylformate decarboxylase, purified to give a single protein band on polyacrylamide gel electrophoresis, was shown to be responsible for the catalysis of this novel condensation reaction.

Thiamin diphosphate in biological chemistry: exploitation of diverse thiamin diphosphate‐dependent enzymes for asymmetric chemoenzymatic synthesis

“…Pp BFD is an exceptional ThDP‐dependent enzyme, because it catalyses the formation of ( S )‐2‐hydroxy‐1‐phenylpropan‐1‐one (HPP) from benzaldehyde and acetaldehyde. This is one of the very few known S ‐selective reactions (92% enantiomeric excess [ ee ]) catalysed by this otherwise R ‐selective class of enzymes [46]. BFD further allows access to various ( S )‐HPP analogues based on the carboligation of acetaldehyde with different aromatic, heteroaromatic, cyclic aliphatic as well as olefinic aldehydes [28,29].…”

Factors Mediating Activity, Selectivity, and Substrate Specificity for the Thiamin Diphosphate‐Dependent Enzymes Benzaldehyde Lyase and Benzoylformate Decarboxylase

“…2 BAL catalyzes the cleavage of aromatic 2‐hydroxy ketones like benzoin, thereby allowing P. fluorescens to grow on media with benzoin as the only carbon source 3. 4 In 1991, Wilcocks and Ward first described a BFD‐catalyzed carboligase reaction that yielded 2‐hydroxypropiophenone (2‐HPP), by using benzoylformate and acetaldehyde as substrates 5. Further studies revealed that both BFD6–12 and BAL9, 13–19 are able to catalyze the carboligation of a broad range of aldehydes; this makes them valuable catalysts for bioorganic syntheses.…”

Carboligation Reactions Mediated by Benzoylformate Decarboxylase Immobilized on a Magnetic Solid Support

“…Wilcocks and Ward reported the carboligase activity of BFD from P. putida for the first time, introducing the catalyst as an efficient alternative for the enantioselective synthesis of (S)-2hydroxyketones. 14 Its stereoselectivity is highly dependent on the structure of the substrate aldehydes. 15 For synthetic purposes, the enzyme also accepts aldehydes as donors, instead of R-ketoacids, to give the corresponding acyloins with (S)-configuration.…”