Biosynthesis of Unnatural Porphyrins in E. Coli

大肠杆菌中非天然卟啉的生物合成

• 批准号: 6623557
• 负责人: CLAUDIA SCHMIDT-DANNERT
• 金额: $ 16.06万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-10 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623557
• 关键词: Escherichia coli; analytical method; bacterial genetics; biological products; biotechnology; chemical structure function; combinatorial chemistry; directed evolution; enzyme mechanism; flow cytometry; gene expression; genetic regulation; high performance liquid chromatography; high throughput technology; method development; microorganism metabolism; molecular cloning; polymerase chain reaction; porphyrin biosynthesis; porphyrins; protein engineering; proteomics; recombinant DNA; spectrometry; thin layer chromatography

The proposed research seeks to combine techniques of metabolic engineering with those of directed enzyme evolution (termed 'molecular pathway breeding') for the protection of useful novel chemical compounds and materials in recombinant cells. This application seek so to expand this approach to the biosynthesis of yet other, even more complex novel chemical compounds-porphyrins-demanding the development of new screening and analytical tools. Porphyrins are a fascinating, structurally complex compound class of enormous significance and potential for a variety of applications in e.g. medicine, chemistry or material sciences. Mimicking natural breeding processes through gene assembly and in vitro evolution of only a few functionally diverse available microbial porphyrin biosynthetic genes will generate new pathways for unnatural porphyrins. The strategies and methods developed in this project can be readily applied to other metabolic pathways and provide new engineering tools for the discovery and production, in recombinant cells, of a kaleidoscope of diverse natural and non-natural compounds for a variety of applications. In particular, the developed high-throughput (HT) screening methods for the identification of synthesized small molecules in large cell libraries will be highly useful for other combinatorial approaches as well. The biosynthetically produced new porphyrin structures with various reactive functional groups will make valuable scaffolds for chemical modifications for the synthesis of additional porphyrin structures.

拟议的研究旨在将代谢工程技术与定向酶进化技术（称为“分子途径育种”）相结合，以保护重组细胞中有用的新型化合物和材料。该应用寻求将这种方法扩展到其他更复杂的新型化合物（卟啉）的生物合成，这需要开发新的筛选和分析工具。卟啉是一类令人着迷、结构复杂的化合物，具有巨大的意义和潜在的多种应用潜力，例如在生物医学领域。医学、化学或材料科学。通过仅少数功能多样的可用微生物卟啉生物合成基因的基因组装和体外进化来模仿自然育种过程，将为非天然卟啉产生新的途径。该项目开发的策略和方法可以很容易地应用于其他代谢途径，并为在重组细胞中发现和生产用于各种应用的各种天然和非天然化合物的万花筒提供新的工程工具。特别是，开发的用于鉴定大细胞文库中合成小分子的高通量（HT）筛选方法对于其他组合方法也非常有用。 生物合成产生的具有各种反应性官能团的新卟啉结构将为合成其他卟啉结构的化学修饰提供有价值的支架。