Fig. 2. Enzyme design for identification of hot-spot residues for TcC (A) and HiC (B). The catalytic triad (S131/H209/D177) of TcC and parts of the oxyanion hole (Y61, M132) are shown. The catalytic triad (S103/H171/D158) of HiC and parts of the oxyanion hole (S26, Q104) are labeled. (C) In silico and experimental evaluation of engineered polyesterases. For reference, the relative abundance of hydrogen bond formation and experimentally determined hydrolysis activities (Table 1) are normalized to wild type in each case. The inset shows a snapshot from an MD-simulation of the TcC I179N variant with the key hydrogen bond indicated by the arrow. The substrate 3PA 6,6 is shown in enlarged sticks and water molecules are labeled “wat”.