BIOSYNTHETIC STUDIES USING 15N-13C CNMR SPECTROSCOPY

使用 15N-13C CNMR 光谱进行生物合成研究

• 批准号: 3280701
• 负责人: STEVEN J GOULD
• 金额: $ 21.68万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-12-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280701
• 关键词: Streptomyces; aminoacid metabolism; aminoglycoside antibiotics; antibiotics; arginine; biological products; biotransformation; carbon; chemical structure function; chloramphenicol; drug design /synthesis /production; genetic manipulation; genome; glucose; hydroxyaminoacid; mass spectrometry; microorganism growth; microorganism metabolism; nitrogen; nuclear magnetic resonance spectroscopy; radiotracer; siderophores; stable isotope; stereochemistry

We propose to continue our studies of antibiotic biosynthesis and will focus on the secondary metabolism of amino acids and of glucose. We will be concerned with the generation of w-N-hydroxy- , 3-hydroxy-, 2,3-dehydro-, and B-amino acids and with the formation and further transformation of nucleosides involved in antibiotic biosynthesis. Key enzymes will be isolated and characterized. The genome for acivicin biosynthesis will be cloned. Experiments are planned to further clarify the biosynthesis of streptothricin F and to determine the biogenetic relationship of capreomycidine (capreomycin biosynthesis) and 3-epicapromycidine (elastatinal biosynthesis). Potential nucleoside intermediates in streptothricin F biosynthesis and in blasticidin S biosynthesis will be synthesized and tested. Arginine-2,3-aminomutase will be isolated from Streptomyces qriseochromogenes (blasticidin S biosynthesis) and its reaction mechanism studied. L-Ornithine N- monooxygenase and acivicin 4-monooxygenase will be isolated from S. sviceus, and the intermediates in acivicin biosynthesis beyond N-hydroxyornithine determined. Studies on the conversion of p- aminophenylalanine to p-aminophenylserine in chloramphenicol biosynthesis (S. 3022a) will be completed. The biosynthesis of cis and trans-L-3-hydroxyproline, of erythro-L-3-hydroxyleucine, of 2,3-dehydrotryptophan, and of threo-L-3-methyltryptophan residues of telomycin will be studied in S. canus. The biosynthesis of threo-D-3-hydroxyaspartic acid and of the N-hydroxy-D-ornithine lactam of the siderophore pseudobactin will be elucidated, as will that of the unusual chromophore of this pseudomonas metabolite. Most of the labeled compounds needed for these studies will be synthesized in our laboratories. We will primarily use stable isotopes with analysis by high field NMR spectroscopic techniques. Our 15N/13C NMR spin coupling technique will continue to be essential for mapping out complex biosynthetic pathways. It is hoped that our investigations will continue to reveal previously obscured biogenetic relationships amongst seemingly diverse structures, and will expand our understanding of organic reactions and reactivity. Our results should lay the groundwork for cloning the genes responsible for production of these antibiotics, and may lead to new, hybrid antibiotics with improved characteristics.

我们建议继续研究抗生素的生物合成， 将集中在氨基酸的次级代谢和 葡萄糖 我们将关注w-N-羟基的生成， ，3-羟基-，2，3-氨基-和B-氨基酸， 参与的核苷的形成和进一步转化 抗生素生物合成 关键酶将被分离， 表征了 用于阿西维辛生物合成的基因组将是 克隆的 计划进行实验以进一步阐明 链丝菌素F，并确定生物遗传关系， 卷曲霉素（卷曲霉素生物合成）和3-表丙霉素 （弹性蛋白生物合成）。 潜在的核苷中间体 链丝菌素F生物合成和杀稻瘟菌素S生物合成 将被合成并测试。 精氨酸-2，3-氨基变位酶将是 分离自灰色产色链霉菌（杀稻瘟菌素S 生物合成），并对其反应机理进行了研究。 L-鸟氨酸 单加氧酶和acivicin 4-单加氧酶将从 S. sviceus，以及除此之外的阿西维辛生物合成中的中间体 测定N-羟基鸟氨酸。 p-的转化研究 氯霉素中氨基苯丙氨酸转化为对氨基苯丝氨酸 生物合成（S. #30220;将完成。 顺式的生物合成 和反式-L-3-羟脯氨酸，红-L-3-羟亮氨酸， 2，3-脱氢色氨酸和苏式-L-3-甲基色氨酸残基 将在S.卡纳斯 的生物合成 苏式-D-3-羟基天冬氨酸和N-羟基-D-鸟氨酸 将阐明铁载体假杆菌素的内酰胺， 这种假单胞菌代谢物中不寻常的发色团。 这些研究所需的大多数标记化合物将是 在我们的实验室合成。 我们将主要使用稳定的 通过高场NMR光谱技术进行同位素分析。 我们的15 N/13 C NMR自旋耦合技术将继续在 对于绘制复杂的生物合成途径至关重要。 希望我们的调查能继续揭示 以前模糊的生物遗传关系， 不同的结构，并将扩大我们对有机的理解， 反应和反应性。 我们的研究结果应该为 用于克隆负责产生这些的基因 抗生素，并可能导致新的，混合抗生素与改善 特色