Characterization of 1,2-Propanediol Dehydratases Reveals Distinct Mechanisms for B₁₂-Dependent and Glycyl Radical Enzymes

Propanediol dehydratase (PD), a recently characterized member of the glycyl radical enzyme (GRE) family, uses protein-based radicals to catalyze the chemically challenging dehydration of (<i>S</i>)-1,2-propanediol. This transformation is also performed by the well-studied enzyme B₁₂-dependent propanediol dehydratase (B₁₂-PD) using an adenosylcobalamin cofactor. Despite the prominence of PD in anaerobic microorganisms, it remains unclear if the mechanism of this enzyme is similar to that of B₁₂-PD. Here we report ¹⁸O labeling experiments that suggest PD and B₁₂-PD employ distinct mechanisms. Unlike B₁₂-PD, PD appears to catalyze the direct elimination of a hydroxyl group from an initially formed substrate-based radical, avoiding the generation of a 1,1-<i>gem</i> diol intermediate. Our studies provide further insights into how GREs perform elimination chemistry and highlight how nature has evolved diverse strategies for catalyzing challenging reactions