The Diiron Monooxygenase CmIA from Chloramphenicol Biosynthesis Allows Reconstitution of β-Hydroxylation during Glycopeptide Antibiotic Biosynthesis
The Diiron Monooxygenase CmIA from Chloramphenicol Biosynthesis Allows Reconstitution of β-Hydroxylation during Glycopeptide Antibiotic Biosynthesis
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DOI：
10.1021/acschembio.9b00862
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发表时间：
2019-12-01
影响因子：
4
通讯作者：
Cryle, Max J.
中科院分区：
文献类型：
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作者：
Kaniusaite, Milda;Goode, Robert J. A.;Cryle, Max J.
β-羟基化在包括博来霉素、氯霉素和糖肽抗生素（GPAs）在内的许多重要天然产物的非核糖体肽生物合成中起重要作用。在这样的过程中涉及各种氧化酶，其中掺入的机制从腺苷酸化之前在氨基酸前体中安装羟基到肽组装期间的直接氨基酸氧化。在这项工作中，我们证明了不寻常的非血红素二铁单加氧酶CmlA从氯霉素生物合成的β-羟基化的各种载体蛋白结合底物的体外效用和范围，通过适应这种酶作为一个非天然的反式作用酶内NRPS介导的GPA生物合成。我们的研究结果表明，CmlA具有广泛的底物特异性的修饰的苯丙氨酸/酪氨酸残基作为底物，并可用于一个实用的策略，功能交叉互补相容NRPS生物合成途径在体外。
beta-Hydroxylation plays an important role in the nonribosomal peptide biosynthesis of many important natural products, including bleomycin, chloramphenicol, and the glycopeptide antibiotics (GPAs). Various oxidative enzymes have been implicated in such a process, with the mechanism of incorporation varying from installation of hydroxyl groups in amino acid precursors prior to adenylation to direct amino acid oxidation during peptide assembly. In this work, we demonstrate the in vitro utility and scope of the unusual nonheme diiron monooxygenase CmlA from chloramphenicol biosynthesis for the beta-hydroxylation of a diverse range of carrier protein bound substrates by adapting this enzyme as a non-native trans-acting enzyme within NRPS-mediated GPA biosynthesis. The results from our study show that CmlA has a broad substrate specificity for modified phenylalanine/tyrosine residues as substrates and can be used in a practical strategy to functionally cross complement compatible NRPS biosynthesis pathways in vitro.