{'fraction_non_alphanumeric': 0.023007395234182416, 'fraction_numerical': 0.007395234182415777, 'mean_word_length': 5.655737704918033, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 0, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
Evolved Aliphatic Halogenases Enable Regiocomplementary C-H Functionalization of an Added-Value Chemical.

Non-heme iron halogenases represent synthetically valuable biocatalysts that are capable of halogenating unactivated sp3-hybridized carbon centers with exquisite stereo- and regioselectivity. The reported substrate scope of these enzymes, however, is limited primarily to the natural substrates and their analogues. Here we engineered the recently discovered halogenase WelO5* for chlorination of a pharmaceutically interesting martinelline-derived fragment. Using structure-guided evolution, a halogenase variant with a more than 290-fold higher total turnover number and a 400-fold higher apparent kcat compared to the wild-type enzyme was generated. Moreover, we identified key positions in the active site which allowed directing the halogen to different positions in our target substrate. This report provides the first example of enzyme engineering to expand the substrate scope of a non-heme iron halogenase beyond the native indole alkaloid-type substrates. The highly evolvable nature of WelO5* underscores the usefulness of this enzyme family for the late-stage halogenation of value-added chemicals.