Molecular dynamics-derived root mean square fluctuation (RMSF) values of residues in protein WT and Y68T (obtained from Alphafold 3)

Our work identifies AmCE1, a ferulic acid esterase from <i>Anaeromyces mucronatus</i>, as a promiscuous hydrolase/acyltransferase—and, to our knowledge, the first of its kind to exhibit unprecedented acyltransferase activity toward bulky aromatic acyl groups in aqueous solution. Concomitant with the rational design of AmCE1 (WT) to Y68T, we have established a sustainable and efficient platform for the industrial production of cinnamate derivatives. Herein, we supplement the Root Mean Square Fluctuation (RMSF) data derived from molecular dynamics simulations, which support the superior transesterification activity of Y68T (modeled by AlphaFold3) compared to the wild type (WT).