ME: Interagency Announcement of Opportunities in Metabolic Engineering: Laboratory Evolution of Carotenoid Biosynthetic Pathways

这项拟议研究的目的是确定类胡萝卜素生物合成如何进化以增加可合成的类胡萝卜素结构的多样性。本研究将从两个层面展开。单个生物合成酶将经历一轮又一轮的突变和重组。精心设计的屏幕将检测到生物合成功能的改变。功能改变的类胡萝卜素生物合成酶将被结合在一起，创造新的途径。通过将氨基酸的变化与生物合成功能的变化联系起来，可以深入了解生物合成功能如何进一步改变，以及自然如何进化途径功能。此外，这项研究还可以为更困难或更复杂的途径的修改提供必要的理解。

The objective of the proposed research is to determine how carotenoid biosynthesis can be evolved to increase the variety of carotenoid structures that can be synthesized. This research will proceed at two levels. Individual biosynthetic enzymes are to be subjected to sequential rounds of mutagenesis and recombination. Carefully designed screens will detect altered biosynthetic function. Carotenoid biosynthetic enzymes with altered functions will then be combined to create new pathways. By correlating amino acid changes with changes in biosynthetic function, insights can be gleaned into how biosynthetic function may be further altered as well as how nature may evolve pathway function. Also, this research could provide the requisite understanding to approach modifications of more difficult or complex pathways.