Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Dr., Urbana, IL 61801, USA.
Department of Chemistry, Purdue University, Herbert C. Brown Laboratory of Chemistry, Room 4103E, 560 Oval Drive, Box 59, West Lafayette, IN 47907, USA.
Angew Chem Int Ed Engl. 2022 Jul 25;61(30):e202206173. doi: 10.1002/anie.202206173. Epub 2022 Jun 7.
The biosynthesis of the natural product dehydrofosmidomycin involves an unusual transformation in which 2-(trimethylamino)ethylphosphonate is rearranged, desaturated and demethylated by the enzyme DfmD, a divergent member of the 2-oxoglutarate-dependent dioxygenase superfamily. Although other members of this enzyme family catalyze superficially similar transformations, the combination of all three reactions in a single enzyme has not previously been observed. By characterizing the products of in vitro reactions with labeled and unlabeled substrates, we show that DfmD performs this transformation in two steps, with the first involving desaturation of the substrate to form 2-(trimethylamino)vinylphosphonate, and the second involving rearrangement and demethylation to form methyldehydrofosmidomycin. These data reveal significant differences from the desaturation and rearrangement reactions catalyzed by other family members.
天然产物去氢土霉素的生物合成涉及一种不寻常的转化，其中 2-(三甲基氨基)乙基膦酸通过酶 DfmD 进行重排、去饱和和去甲基化，DfmD 是 2-酮戊二酸依赖性双加氧酶超家族的一个分支成员。尽管该酶家族的其他成员催化表面上相似的转化，但在单个酶中同时进行这三个反应以前尚未观察到。通过对标记和未标记底物的体外反应产物进行表征，我们表明 DfmD 分两步完成此转化，第一步涉及底物的去饱和以形成 2-(三甲基氨基)乙烯基膦酸，第二步涉及重排和去甲基化以形成甲基金去氢土霉素。这些数据与其他家族成员催化的去饱和和重排反应有显著差异。
