Biosynthesis of Unnatural Porphyrins in E. Coli

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化合物和材料。本应用程序试图将这种方法扩展到其他更复杂的新型化合物（卟啉）的生物合成中，这需要开发新的筛选和分析工具。卟啉是一种令人着迷的、结构复杂的化合物，在医学、化学或材料科学等领域具有巨大的意义和潜力。通过基因组装模拟自然育种过程和体外进化只有少数功能多样的微生物卟啉生物合成基因将为非天然卟啉提供新的途径。本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，并为重组细胞中各种天然和非天然化合物的万花筒的发现和生产提供新的工程工具，用于各种应用。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）筛选方法也将对其他组合方法非常有用。生物合成的具有各种活性官能团的新的卟啉结构将为化学修饰合成其他卟啉结构提供有价值的支架。

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.