Biosynthesis of Unnatural Porphyrins in E. Coli

这项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。这项应用寻求将这种方法扩展到生物合成其他更复杂的新化合物--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迷人的化合物，在医学、化学或材料科学等领域具有巨大的意义和潜在的应用前景。通过基因组装和体外进化来模拟自然繁殖过程，只有几个功能不同的可用的微生物卟啉生物合成基因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各种天然和非天然化合物的万花筒提供新的工程工具，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。生物合成的具有不同活性官能团的新的卟啉结构将为合成额外的卟啉结构提供有价值的化学修饰支架。

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.