Prenylated flavin mononucleotide (prFMN) is a recently discovered cofactor required by the UbiD family of reversible decarboxylases involved in ubiquinone biosynthesis, biological decomposition of lignin, and biotransformation of aromatic compounds. This cofactor is synthesized by UbiX-like prenyltransferases catalyzing the transfer of the dimethylallyl moiety of dimethylallyl-monophosphate (DMAP) to FMN. The origin of DMAP for prFMN biosynthesis and the biochemical properties of free prFMN are unknown. We show that inEscherichia colicells, DMAP can be produced by phosphorylating prenol using ThiM or dephosphorylating DMAPP using Nudix hydrolases. We produced 14 active prenyltransferases whose properties enabled the purification and characterization of protein-free forms of prFMN.In vitroassays revealed that the UbiD-like ferulate decarboxylase (Fdc1) can be activated by free prFMNiminiumor C2′-hydroxylated prFMNiminiumunder both oxidized and reduced conditions. These insights into the biosynthesis and properties of prFMN will facilitate further elucidation of the biochemical diversity of reversible UbiD (de)carboxylases.

中文翻译：

---

烯丙基化FMN辅因子的生物合成和活性

泛烯基黄素单核苷酸（prFMN）是UbiD可逆脱羧酶家族中需要的辅因子，该因子参与泛醌生物合成，木质素的生物分解和芳族化合物的生物转化。该辅助因子是通过UbiX样异戊二烯基转移酶合成的，该酶催化将二甲基烯丙基单磷酸酯（DMAP）的二甲基烯丙基部分转移至FMN。prFMN生物合成的DMAP的起源和游离prFMN的生化特性尚不清楚。我们显示在大肠杆菌细胞中，可以通过使用ThiM磷酸化烯丙醇或使用Nudix水解酶使DMAPP脱磷酸来产生DMAP。我们生产了14种活性异戊烯基转移酶，其性质能够纯化和表征无蛋白形式的prFMN。体外测定表明，在氧化和还原条件下，游离prFMNiminium或C2'-羟基化prFMNiminium均可激活UbiD样阿魏酸脱羧酶（Fdc1）。这些对prFMN的生物合成和性质的见解将有助于进一步阐明可逆UbiD（de）羧化酶的生物化学多样性。