BIOSYNTHETIC STUDIES USING 15N 13C CNMR SPECTROSCOPY

我们建议继续进行抗生素生物合成的研究， 专注于氨基酸和葡萄糖的次级代谢。 与 前者，我们的重点将是β-羟基-α-氨基的产生 氨基酸和β-氨基酸作为生物合成中间体。 这项工作 将包括对链丝菌素F，杀稻瘟菌素， 阴霉素和3-羟阴霉素，并将扩大到包括 生物遗传学相关代谢物弹性蛋白，AT-125，和 氯霉素，以测试我们的假设的形成和 β-羟基氨基酸的利用。 我们将主要使用稳定的 同位素13 C、2 H、15 N和18 O，它们将通过NMR检测 光谱技术 一些标记的化合物， 将在其中一个以上的项目中使用。 同位素标记模式的分析将再次使用我们的15 N/13 C NMR 自旋耦合技术，将探索我们的新的1H/2 H COSY的实用性 技术编辑复杂的2 H NMR谱，并将探讨实用 3 H NMR和高分辨率FAB质谱用于定位同位素 复杂代谢物中的标记。 我们的调查将增加我们对哪种化学物质的了解 反应在自然界中是可用的，以及它们如何被连接到生产 复杂的结构。 它们还可以提供产生新的 类似物，也具有抗生素活性， 生物合成装置使用改性的前体。

We propose to continue our studies of antibiotic biosynthesis, and will focus on the secondary metabolism of amino acids and of glucose. With the former our emphasis will be on the generation of Beta-hydroxy-Alpha-amino acids and of Beta-amino acids as biosynthetic intermediates. This work will include further experiments on streptothricin F, blasticidin, negamycin and 3-epideoxynegamycin, and will be expanded to include the biogenetically relevant metabolites elastatinal, AT-125, and chloramphenicol in order to test our hypotheses on the formation and utilization of Beta-hydroxy amino acids. We will primarily use the stable isotopes 13C, 2H, 15N, and 18O and they will be detected by NMR spectroscopic techniques. A number of labeled compounds that will be synthesized will be used in more than one of these projects. Analysis of the isotope labeling patterns will again use our 15N/13C NMR spin coupling technique, will explore the utility of our new 1H/2H COSY technique for editing complex 2H NMR spectra, and will explore the utility of 3H NMR and of high resolution FAB mass spectrometry for locating isotope labels in complex metabolites. Our investigations will increase our understanding of which chemical reactions are available in Nature and how they can be linred to produce complex structures. They may also provide the opportunity to produce new analogs, also with antibiotic activity, by redirecting the microorganisms' biosynthetic apparatus to use modified precursors.