FRIES Alexander Erich
Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
HAMPEL, SABRINA; STEITZ, JAN-PATRICK; BAIERL, ANNA; LEHWALD, PATRIZIA; WIESLI, LUZIA; RICHTER, MICHAEL; FRIES, ALEXANDER; POHL, MARTINA; SCHNEIDER, GUNTER; DOBRITZSCH, DOREEN; MÜLLER, MICHAEL
WILEY-V C H VERLAG GMBH
Año: 2018 vol. 19 p. 2283 - 2283
A wide range of thiamine diphosphate (ThDP)-dependent enzymes catalyze the benzoin-type carboligation of pyruvate with aldehydes. A few ThDP-dependent enzymes, such as YerE from Yersinia pseudotuberculosis (YpYerE), are known to accept ketones as acceptor substrates. Catalysis by YpYerE gives access to chiral tertiary alcohols, a group of products difficult to obtain in an enantioenriched form by other means. Hence, knowledge of the three-dimensional structure of the enzyme is crucial to identify structure?activity relationships. However, YpYerE has yet to be crystallized, despite several attempts. Herein, we show that a homologue of YpYerE, namely, PpYerE from Pseudomonas protegens (59 % amino acid identity), displays similar catalytic activity: benzaldehyde and its derivatives as well as ketones are converted into chiral 2-hydroxy ketones by using pyruvate as a donor. To enable comparison of aldehyde- and ketone-accepting enzymes and to guide site-directed mutagenesis studies, PpYerE was crystallized and its structure was determined to a resolution of 1.55 Å.