BIOSYNTHESIS OF MACROLIDE ANTIBIOTICS

大环内酯类抗生素的生物合成

• 批准号: 3280348
• 负责人: CHARLES R HUTCHINSON
• 金额: $ 13.84万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1992-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280348
• 关键词: Streptomyces; antiinfective agents; bacterial genetics; biotechnology; erythromycin; gene expression; genetic mapping; genetic promoter element; genetic transcription; genetic translation; macrolide antibiotics; microorganism culture; microorganism metabolism; mutant; nucleic acid sequence; plasmids

This proposal requests four years of non-competing support for a study of the biochemistry and genetics of erythromycin A production by Streptomyces erythreus and midecamycin production by Streptomyces mycarofaciens. These macrolide antibiotics are widely used antiinfective agents; thus information about the genetic and biochemical mechanisms controlling their biosynthesis would have important benefits to the applied sciences. Their formation involves over twenty enzymatically catalyzed steps and perhaps as many structural genes. This suggests that a study of the organization and expression of the ery and mid production genes should uncover complex and interesting genetic and biochemical control mechanisms. We propose to establish the organization of the genes that control deoxysugar biosynthesis and the hydroxylation of a macrolactone intermediate in the erythromycin pathway, and to analyze the factors controlling their expression. In this work, we will investigate two interesting research leads found in the previous grant period which point to the existence of novel transcriptional or translational control mechanisms. The experiments will involve gentic complementation, DNA sequencing, promoter identification, and the analysis of transcriptional organization by low and high resolution S1 mapping and Northern hybridization. Expression of these genes as a function of time and growth conditions will be measured by dot-blot experiments. The characterization three enzymes encoded by these genes also will be pursued. The genes governing the assembly of platenolide I, an early intermediate of the midecamycin pathway, also will be studied. We will isolate a collection of Mid mutants of S. mycarofaciens and use them to locate mid genes by complementation of midA mutations and mutational cloning or transposon mutagenesis. A new method for characterizing the biochemical deficiency in MidA mutants also will be evaluated.

该提案要求四年的非竞争支持 红霉素A的生物化学和遗传学研究 链霉菌和中霉素产生的生产 由链霉菌霉菌。 这些大环内酯类抗生素是 广泛使用的抗探测剂；因此有关 控制其生物合成的遗传和生化机制 将对应用科学有重要好处。 他们的 形成涉及二十多个酶促催化的步骤和 也许是许多结构基因。 这表明对 ERY和中期生产的组织和表达 基因应发现复杂而有趣的遗传和 生化控制机制。 我们建议建立控制基因的组织 脱氧生物合成和大分子的羟基化 在红霉素途径中中间，并分析 控制其表达的因素。 在这项工作中，我们将 调查上一个有趣的研究领导 赠款时期，表明存在新颖的转录 或翻译控制机制。 实验会 涉及绅士互补，DNA测序，启动子 识别以及对转录组织的分析 低分辨率和高分辨率S1映射和北部杂交。 这些基因作为时间和生长的函数的表达 条件将通过点印刷实验来衡量。 这 表征这些基因编码的三种酶也将 被追捕。 铂醇i的组装的基因，早期 还将研究中霉素途径的中级。 我们将隔离菌霉菌的中间突变体的集合 并用它们通过互补MIDA来定位中基因 突变和突变克隆或转座子诱变。 一个 表征生化缺陷的新方法 也将评估MIDA突变体。