Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA.
Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, USA.
Angew Chem Int Ed Engl. 2018 Jul 2;57(27):8048-8052. doi: 10.1002/anie.201802856. Epub 2018 Jun 12.
The bioactivity of vancomycin is enabled by three aromatic crosslinks, the biosynthesis of which has been an active area of investigation for two decades. Two cytochrome P450 enzymes, OxyB and OxyA, have been shown to introduce bisaryl ether linkages with the help of a so-called X-domain. The final crosslink, however, a biaryl bond thought to be installed by OxyC, has remained elusive. We report the in vitro reconstitution of the OxyC reaction and formation of the first carbon-carbon crosslink in any glycopeptide antibiotic. Using a cascade sequence, in which the peptide substrate was incubated with the Oxy enzymes in turn, we completed the chemoenzymatic synthesis of a vancomycin aglycone variant. This approach was also used to generate a new analogue carrying a thioamide linkage at residue 4, a precursor to the amidine derivative, which is effective against vancomycin-resistant pathogens. Our results set the stage for creating therapeutic vancomycin derivatives by using the native metalloenzymes.
万古霉素的生物活性得益于三个芳族交联键，其生物合成是过去二十年研究的热点。两种细胞色素 P450 酶（OxyB 和 OxyA）在所谓的 X 结构域的帮助下引入双芳基醚键。然而，最后一个交联键，即被认为由 OxyC 安装的二芳基键，仍然难以捉摸。我们报告了 OxyC 反应的体外重建以及任何糖肽抗生素中第一个碳-碳交联键的形成。使用级联序列，我们依次将肽底物与 Oxy 酶孵育，完成了万古霉素糖苷配基变体的化学酶合成。这种方法还用于生成一种在残基 4 处带有硫代酰胺键的新类似物，这是针对耐万古霉素病原体的脒衍生物的前体。我们的研究结果为使用天然金属酶创造治疗性万古霉素衍生物奠定了基础。
