Rapid access to glycopeptide antibiotic precursor peptides coupled with cytochrome P450-mediated catalysis: towards a biomimetic synthesis of glycopeptide antibiotics
Rapid access to glycopeptide antibiotic precursor peptides coupled with cytochrome P450-mediated catalysis: towards a biomimetic synthesis of glycopeptide antibiotics
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DOI：
10.1039/c4ob02452d
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Cryle, Max J.
中科院分区：
文献类型：
--
作者：
Brieke, Clara;Kratzig, Veronika;Cryle, Max J.
了解糖肽抗生素生物合成的基础机制是未来重新发明这些化合物的关键。为了有效地在体外表征生物合成的关键后期步骤，容易获得广泛的底物肽作为其辅酶A（CoA）缀合物是必不可少的。在这里，我们报告了糖肽前体CoA缀合物的快速路线的发展，提供了高产率和优良的纯度。该合成路线适用于合成含有易外消旋的芳基甘氨酸残基的肽CoA-缀合物：这些残基是非核糖体肽合成的标志，并且以前无法使用Fmoc型化学进行肽合成。我们应用这一途径以载体蛋白结合形式产生糖肽前体肽作为底物，以探索万古霉素生物合成中第一种加氧酶（OxyBvan）的特异性。我们的研究结果表明，OxyBvan是一种高度混杂的催化剂，用于不同糖肽前体的酚偶联，其接受多种载体蛋白底物，甚至在来自替代糖肽生物合成机器的载体蛋白结构域上。这些结果代表了第一个重要的步骤，在体外仿生合成的改性糖肽糖苷配基的发展。
Understanding the mechanisms underpinning glycopeptide antibiotic biosynthesis is key to the future ability to reinvent these compounds. For effective in vitro characterization of the crucial later steps of the biosynthesis, facile access to a wide range of substrate peptides as their Coenzyme A (CoA) conjugates is essential. Here we report the development of a rapid route to glycopeptide precursor CoA conjugates that affords both high yields and excellent purities. This synthesis route is applicable to the synthesis of peptide CoA-conjugates containing racemization-prone arylglycine residues: such residues are hallmarks of non-ribosomal peptide synthesis and have previously been inaccessible to peptide synthesis using Fmoc-type chemistry. We have applied this route to generate glycopeptide precursor peptides in their carrier protein-bound form as substrates to explore the specificity of the first oxygenase enzyme from vancomycin biosynthesis (OxyBvan). Our results indicate that OxyBvan is a highly promiscuous catalyst for phenolic coupling of diverse glycopeptide precursors that accepts multiple carrier protein substrates, even on carrier protein domains from alternate glycopeptide biosynthetic machineries. These results represent the first important steps in the development of an in vitro biomimetic synthesis of modified glycopeptide aglycones.