Tuning of pK_a values activates substrates in flavin-dependent aromatic hydroxylases.

ABSTRACT: Hydroxylation of substituted phenols by flavin-dependent monooxygenases is the first step of their biotransformation in various microorganisms. The reaction is thought to proceed via electrophilic aromatic substitution, catalyzed by enzymatic deprotonation of substrate, in single-component hydroxylases that use flavin as a cofactor (group A). However, two-component hydroxylases (group D), which use reduced flavin as a co-substrate, are less amenable to spectroscopic investigation. Herein, we employed ¹⁹F NMR in conjunction with fluorinated substrate analogs to directly measure pK_a values and to monitor protein events in hydroxylase active sites. We found that the single-component monooxygenase 3-hydroxybenzoate 6-hydroxylase (3HB6H) depresses the pK_a

SUBMITTER: Pitsawong W 

PROVIDER: S-EPMC7086025 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature