Structural Biology of Biosynthetic Enzymes

旨在发现和表征新型磷酸盐的方案项目目标大大受益于生物化学和结构生物学特性以及参与这些天然产物生物合成的酶。此外，详细了解这些酶与它们的同源底物/抑制剂形成的复合体的三维结构有助于设计这些催化剂，以产生具有改进的生物和/或药代动力学性质的衍生物。在下一个周期中，我们将研究目标扩展到生化 抗菌白霉素和抗真菌根霉素生物合成相关基因簇的特征。来自这些簇的酶指导两种天然产品的生产，其中包括一种常见的弹头(苏氨酸合成酶抑制剂(Z)-L-2-氨基-5-膦-3-戊烯酸(APPA))，但使用不同的基于多肽的输送工具。我们将从这些簇中的每一个进行单个酶的体外重组，并利用这一知识来生产APPA的附加多肽和非肽衍生物，这些衍生物可以靶向一系列 其他病原生物。同时，我们的目标也是继续我们对几种生物合成酶的结构和功能的研究，这些酶已经在最初的周期中得到了表征。最后，我们还将描述产生菌利用的抗药性机制，并可能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 oif the enzymes involved in the biosynthesis of these natural products. Additionally, detailed knowledge of the three dimensional 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 producfion of addifional pepfidic 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.