Improving Toluene Dioxygenase Activity for Ester‐Functionalized Substrates through Enzyme Engineering

Rieske dioxygenases have a history of utility in organic synthesis, owing to their ability catalyze the asymmetric dihydroxylation aromatics produce chiral diene-diol metabolites. However, as green-chemical tools has been limited by steric and electronic constraints on substrate scopes activity. Herein we report rational engineering widely used dioxygenase, toluene dioxygenase (TDO), improve activity this enzyme system for synthetically valuable class which wild-type possesses low activity, ester-functionalized aromatics. Through active site targeted mutagenesis application recently reported high throughput screening platform, engineered TDO variants with significantly increased these substrates were identified characterized, revealing key residues that modulate enzyme's selectivity.