Structure-guided engineering of a flavin-containing monooxygenase for the efficient production of indirubin.

ABSTRACT: Indirubin is a bisindole compound for the treatment of chronic myelocytic leukemia. Here, we presented a structure-guided method to improve the activity of a flavin-containing monooxygenase (bFMO) for the efficient production of indirubin in Escherichia coli. A flexible loop interlocked with the active pocket through a helix and the substrate tunnel rather than the active pocket in bFMO were identified to be two reconfigurable structures to improve its activity, resulting in K223R and N291T mutants with enhanced catalytic activity by 2.5- and 2.0-fold, respectively. A combined modification at the two regions (K223R/D317S) achieved a 6.6-fold improvement in catalytic efficiency (k_cat/K_m) due to enhancing π-π stacking interactions stabilization. Finally, an engineered E

SUBMITTER: Sun BY 

PROVIDER: S-EPMC10991670 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature