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Biocatalytic and FMN Prenylation and (De)carboxylase Activation. | LitMetric
Reversible UbiD-like (de)carboxylases represent a large family of mostly uncharacterized enzymes, which require the recently discovered prenylated FMN (prFMN) cofactor for activity. Functional characterization of novel UbiDs is hampered by a lack of robust protocols for prFMN generation and UbiD activation. Here, we report two systems for and FMN prenylation and UbiD activation under aerobic conditions. The one-pot prFMN cascade includes five enzymes: FMN prenyltransferase (UbiX), prenol kinase, polyphosphate kinase, formate dehydrogenase, and FMN reductase, which use prenol, polyphosphate, formate, ATP, NAD, and FMN as substrates and cofactors. Under aerobic conditions, this cascade produced prFMN from FMN with over 98% conversion and activated purified ferulic acid decarboxylase Fdc1 from and protocatechuic acid decarboxylase ENC0058 from . The system for FMN prenylation and UbiD activation is based on the coexpression of Fdc1 and UbiX in cells under aerobic conditions in the presence of prenol. The and FMN prenylation cascades will facilitate functional characterization of novel UbiDs and their applications.
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