Structural Biology of Biosynthetic Enzymes

该计划项目的目标是发现和表征新型的磷酸盐 从酶的生化和结构生物学特性中受益匪浅 参与这些天然产物的生物合成。此外，详细的三维知识 这类酶与其同源底物/抑制剂的复合体结构可以帮助 这些催化剂的工程设计，以产生具有改进的生物和/或 药代动力学特性。在下一个周期中，我们将研究目标扩展到生化 抗菌药物白霉素生物合成相关基因簇的研究 抗真菌根霉素。来自这些簇的酶指导着两种天然产品的生产 由普通弹头(苏氨酸合成酶抑制剂(Z)-L-2-氨基-5-膦-3- 戊烯酸(APPA)，但以多肽为基础的递送载体不同。我们将在体外进行 从这些簇中的每一个中重组单独的酶并利用这一知识 生产额外的APPA的多肽和非多肽衍生物，可以针对一系列 其他病原生物。同时，我们还致力于继续我们的结构-功能研究 在最初的循环中已经被表征的几种生物合成酶。最后，我们 也将表征产生生物利用的抗性机制和 可能会限制几种磷酸盐的生物用途。

The Program Project goals towards the discovery and characterization of novel phosphonates have benefited significantly from biochemical and structural biological characterization of the enzymes involved in the biosynthesis of these natural products. Additionally, detailed knowledge of the threedimensional structures of such enzymes in complex with their cognate substrates/inhibitors can aid in the engineering of these catalysts to yield derivative compounds with improved biological and/or pharmacokinetic properties. In this next cycle, we expand our research aims towards the biochemical characterization of gene clusters involved in the biosynthesis of the antibacterial plumbemycin and the antifungal rhizoctlcin. The enzymes from these clusters direct the production of two natural products consisting of a common warhead (the threonine synthase inhibitor (Z)-L-2-amino-5-phosphono-3- pentenoic acid (APPA)) but with different peptide-based delivery vehicles. We will carry out in vitro reconstitution of individual enzymes from each of these clusters and utilize this knowledge for the production of additional peptidic and non-peptidic derivatives of APPA that can target a range of additional pathogenic organisms. Concurrently, we also aim to continue our structure-function studies of several of the biosynthetic enzymes that have been characterized during the initial cycle. Lastly, we will also characterize the mechanisms of resistance that are utilized by the producing organisms and may limit the biological utility of several phosphonates.