Sequential In Vitro Cyclization by Cytochrome P450 Enzymes of Glycopeptide Antibiotic Precursors Bearing the X-Domain from Nonribosomal Peptide Biosynthesis
Sequential In Vitro Cyclization by Cytochrome P450 Enzymes of Glycopeptide Antibiotic Precursors Bearing the X-Domain from Nonribosomal Peptide Biosynthesis
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DOI：
10.1002/anie.201507533
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发表时间：
2015-12-21
影响因子：
16.6
通讯作者：
Cryle, Max J.
中科院分区：
文献类型：
--
作者：
Brieke, Clara;Peschke, Madeleine;Cryle, Max J.
糖肽抗生素（包括万古霉素和替考拉宁）的生物合成依赖于产生肽的非核糖体肽合成酶 (NRPS) 和催化肽侧链交联的细胞色素 P450 酶 (P450s) 之间的相互作用。我们证明，通过使用 NRPS 肽载体蛋白 (PCP)-X 双结构域作为 P450 招募平台，可以实现体外 P450 催化的肽底物连续环化。这项研究表明，虽然前体肽序列影响 P450 OxyA(tei) 的第二个交联的安装，但活性并不限于天然替考拉宁肽。替考拉宁和万古霉素 OxyB 同系物可以进行初始肽环化，并且后者在测试的所有底物组合中均表现出优异的活性。通过使用非天然 X 结构域底物，还显示了六肽的双环化，这证明了该方法在体外环化各种肽底物的效用。
The biosynthesis of the glycopeptide antibiotics, which include vancomycin and teicoplanin, relies on the interplay between the peptide-producing non-ribosomal peptide synthetase (NRPS) and Cytochrome P450 enzymes (P450s) that catalyze side-chain crosslinking of the peptide. We demonstrate that sequential in vitro P450-catalyzed cyclization of peptide substrates is enabled by the use of an NRPS peptide carrier protein (PCP)-X di-domain as a P450 recruitment platform. This study reveals that whilst the precursor peptide sequence influences the installation of the second crosslink by the P450 OxyA(tei), activity is not restricted to the native teicoplanin peptide. Initial peptide cyclization is possible with teicoplanin and vancomycin OxyB homologues, and the latter displays excellent activity with all substrate combinations tested. By using non-natural X-domain substrates, bicyclization of hexapeptides was also shown, which demonstrates the utility of this method for the cyclization of varied peptide substrates in vitro.