Extended Data Fig. 7: Comparison of crystal structures and features of HotPETase and IsPETaseTS.
From: Directed evolution of an efficient and thermostable PET depolymerase
(a) A global superposition of IsPETaseTS (yellow) and HotPETase (light blue). Mutations in IsPETaseTS compared to IsPETaseWT are highlighted with yellow spheres. Mutations installed during directed evolution are highlighted with pink spheres. The rationally inserted disulphide bridge is highlighted with black spheres. The catalytic triad and W185 are in ball and stick representation coloured by all atoms, with blue and grey carbon atoms, respectively. (b) The disulphide bridge is correctly formed between C233 and C282 in HotPETase. Electron density is 2Fo-Fc contoured at 1 sigma (blue) and 2 sigma (yellow). (c) The conversion of P181 in IsPETaseTS to V181 in HotPETase (highlighted pink) results in extension of β-sheet 6, and the formation of an additional hydrogen bond (dashed lines) to L199. (d) In HotPETase, the wobbling tryptophan (W185, grey sticks), forms a π-stacking interaction (dashed line) with the installed Y214 (pink sticks).
