Assessing the Thiamine Diphosphate Dependent Pyruvate Dehydrogenase E1 Subunit for Carboligation Reactions with Aliphatic Ketoacids.

The synthetic properties of the Thiamine diphosphate (ThDP)-dependent pyruvate dehydrogenase E1 subunit from (PDH E1) was assessed for carboligation reactions with aliphatic ketoacids. Due to its role in metabolism, PDH E1 was previously characterised with respect to its biochemical properties, but it was never applied for synthetic purposes. Here, we show that PDH E1 is a promising biocatalyst for the production of chiral α-hydroxyketones. WT PDH E1 shows a 180-250-fold higher catalytic efficiency towards 2-oxobutyrate or pyruvate, respectively, in comparison to engineered transketolase variants from (TK). Its broad active site cleft allows for the efficient conversion of both ()- and ()-configured α-hydroxyaldehydes, next to linear and branched aliphatic aldehydes as acceptor substrates under kinetically controlled conditions. The alternate, thermodynamically controlled self-reaction of aliphatic aldehydes was shown to be limited to low levels of conversion, which we propose to be due to their large hydration constants. Additionally, the thermodynamically controlled approach was demonstrated to suffer from a loss of stereoselectivity, which makes it unfeasible for aliphatic substrates.