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: 3280699
• 负责人: STEVEN J GOULD
• 金额: $ 19.55万
• 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 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280699
• 关键词: Streptomyces; aminoacid metabolism; aminoglycoside antibiotics; antibiotics; arginine; biotransformation; chemical structure function; chloramphenicol; drug design /synthesis /production; genome; glucose; hydroxyaminoacid; mass spectrometry; microorganism growth; microorganism metabolism; nuclear magnetic resonance spectroscopy; radiotracer; siderophores; 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和 将专注于氨基酸的次级新陈代谢和 葡萄糖。我们将关注w-N-羟基的生成- 、3-羟基、2，3-脱氢和B-氨基酸，并与 核苷类化合物的形成和进一步转化 抗生素的生物合成。将分离关键酶并 特色化的。阿昔维菌素生物合成的基因组将是 克隆的。 计划进行实验，以进一步阐明生物合成 链霉菌素F和确定其生物发生关系 卷曲菌素(卷曲霉素生物合成)和3-表丙菌素 (弹性生物合成)。潜在的核苷中间体 链霉菌素F的生物合成及杀菌素S的生物合成 将被合成和测试。精氨酸-2，3-氨基变位酶 拟变色链霉菌(杀菌素S)的分离 生物合成)及其反应机理进行了研究。L-鸟氨酸N- 单加氧酶和酸性蛋白4单加氧酶将从 S.sviceus，以及Acvicin生物合成的中间体 测定了N-羟基鸟氨酸。对苯二酚转化率的研究 氯霉素中氨基苯丙氨酸合成对氨基苯丝氨酸 生物合成(S.3022a)将完成。顺式化合物的生物合成 和反式-L-3-羟基赖氨酸，红血球-L-3-羟基亮氨酸， 2，3-脱氢色氨酸和苏氨酸-L-3-甲基色氨酸残基 将在犬链霉菌中进行研究。……的生物合成 苏氨酸和N-羟基-D-鸟氨酸 铁载体假细菌蛋白的内酰胺也将被阐明。 这种假单胞菌代谢产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的。我们将主要使用稳定的 用高场核磁共振光谱技术进行分析的同位素。 我们的15N/13C核磁共振自旋耦合技术将继续 对绘制复杂的生物合成途径至关重要。 希望我们的调查将继续揭示 以前看起来模糊的生物遗传关系 多样化的结构，并将扩大我们对有机的理解 反应和反应性。我们的结果应该会奠定基础 用于克隆负责生产这些病毒的基因 抗生素，并可能导致新的，经过改进的混合抗生素 特点。