Combinatorial biosynthetic libraries from cyanobacteria

蓝细菌的组合生物合成文库

• 批准号: 6300682
• 负责人: WILLIAM H GERWICK
• 金额: $ 4.9万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-10 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300682
• 关键词: Streptomyces; antineoplastics; biological products; combinatorial chemistry; drug discovery /isolation; genetic library; high performance liquid chromatography; microorganism culture; microorganism metabolism; photosynthetic bacteria; protein engineering; transfection

The overall goal of this Laboratory Program is to produce libraries of natural product-derived substances by innovative methods and to evaluate these in "Smart" bioassay systems targeted to cancer. Despite intensive effort in many fields of science and medicine to find effective treatments for this complex family of diseases, the incidence and morbidity due to cancer continues to look very bleak. This Laboratory Program will focus on the unique chemistries of marine cyanobacteria, and will integrate with the efforts of the other laboratory programs of this Program Project Grant application to clone natural products biosynthetic genes and over express these in a convenient host organism, Streptomyces venezuelae. Four parallel tracks will be taken in this pursuit: 1) our laboratory will provide cultured marine cyanobacteria to Laboratory Program #1 for genomic DNA insolation and subsequent combinatorial biology efforts in Laboratory Program #2; 2) our laboratory will provide information to Laboratory Program #2 on biosynthetic pathways and partial protein sequences of key "tailoring" enzymes to aid in the cloning of genes of specific marine cyanobacterial metabolites of high relevance to cancer and diversity generation; 3) our laboratory will integrate with Novartis "Smart" assays (Laboratory Program #3) to isolate and structurally define new lead compounds deriving from the above integrated molecular biological approaches; and 4) our laboratory will use unique tailoring enzymes cloned from marine cyanobacteria and modified by protein engineering techniques to biotransform additional natural product structures of cancer relevance. Diode-array HPLC methods will be used to dereplicate parent structures, known compounds, and nuisance substance in the combinatorial biosynthetic libraries showing activity in the Novartis "Smart" assays.

该实验室计划的总体目标是通过创新方法产生天然产物衍生物质库，并在针对癌症的“智能”生物测定系统中对其进行评估。尽管许多科学和医学领域都在加紧努力寻找治疗这一复杂疾病家族的有效方法，但癌症的发病率和发病率仍然看起来非常黯淡。该实验室计划将专注于海洋蓝藻的独特化学，并将与本计划项目资助申请的其他实验室计划的努力相结合，以克隆天然产物生物合成基因，并在方便的宿主生物体中过表达这些基因，委内瑞拉链霉菌。在这方面，我们将采取四个平行的轨道：1）我们的实验室将提供培养的海洋蓝细菌的实验室计划#1的基因组DNA分离和随后的组合生物学实验室计划#2的努力; 2）我们的实验室将为实验室计划#2提供关于生物合成途径和关键“剪裁”的部分蛋白质序列的信息。有助于克隆与癌症和多样性产生高度相关的特定海洋蓝藻代谢物的基因的酶; 3）我们的实验室将与诺华的“智能”分析相结合（实验室程序#3）分离和结构上确定从上述综合分子生物学方法衍生的新的先导化合物;以及4）我们的实验室将使用从海洋蓝细菌中克隆并通过蛋白质工程技术修饰的独特定制酶，生物转化与癌症相关的其他天然产物结构。将使用二极管阵列HPLC方法对在Novartis“Smart”试验中显示活性的组合生物合成文库中的母体结构、已知化合物和有害物质进行去复制。