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型化合物和材料。本申请寻求将这种方法扩展到生物合成其他甚至更复杂的新型化学化合物-卟啉-需要开发新的筛选和分析工具。卟啉是一类结构复杂的化合物，在医学、化学或材料科学等领域具有广泛的应用前景。通过基因组装和体外进化来模拟自然繁殖过程，只有少数功能多样的微生物卟啉生物合成基因将产生新的非天然卟啉途径。该项目中开发的策略和方法可以很容易地应用于其他代谢途径，并提供新的工程工具，用于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。特别是，开发的高通量（HT）筛选方法，用于在大细胞库中的合成的小分子的鉴定将是非常有用的其他组合的方法，以及。 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额外的卟啉结构的化学修饰提供有价值的支架。

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.