Evolving Old Yellow Enzymes OYEs

The origin of the proton abstracted by the formed carbanion was also determined by site-specific mutagenesis. Several studies showed that exchanges of a strongly conserved tyrosine residue to phenylalanine or alanine leads to an overall reduced activity [8-10]. In contrast to OYEl [8], the studies with the OYEs designated as morphinone reductase (MR) and pentaerythritol tetranitrate (PETN) suggest that solvent water is the source of the proton required for reduction [9,10]. 

Positions in space confirmed as hot spots are identified by structural analysis of OYE family highlighted as green sticks residues involved in members. The FMN cofactor is highlighted in substrate binding and proton donation are orange and bound molecules in yellow. YqjM 

Evolving OYEl as a Catalyst in the Stereoselective Reduction of 3 Alkyl-2-cyclo-hexenone Derivatives and Baylis-Hillman Adducts 

An early example of protein engineering of an OYE pertains to the reductase from S. pastorianus as a catalyst in the stereoselective reduction of hulky 3-aIkyl-suhstituted 2-cyclohexenone derivatives in the presence of an NADPH regeneration system [35]. 

PDB entry 3TX9 3UPW 3P7Y 1Z44 3N19 4A3U 

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Evolving Old Yellow Enzymes (OYEs) 1121 Enzyme Conversion (%) ee (%)... 

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