Structural Elucidation and Biosynthesis

在之前的资助期内，我们的酶学和合成化学研究补充了遗传和生化方面的努力，以了解天然产物磷酸盐的生物合成逻辑。该项目团队已经取得了重大进展，开发了有效的方法来检测新的膦酸盐生物合成基因簇，并非常详细地了解了生物合成途径，这些途径导致了重要的商业膦酸盐，如磷丙氨酸和fostomycin。随着遗传和生化研究效率的提高，一个新的瓶颈被发现，即涉及低效的纯化策略，从而减缓了新发现的基因簇产物的结构解析。本分项目的部分拟议研究将侧重于开发有效的净化和

In the previous grant period, our enzymology and synthetic chemistry studies complemented the genetic and biochemical efforts towards understanding the biosynthetic logic of natural product phosphonates. The program project team has made significant progress, developing effective methods to detect new phosphonate biosynthetic gene clusters and understanding in great detail the biosynthetic pathways leading towards commercially important phosphonates such as phosphinothricin and fostomycin. As the genetic and biochemical studies became more efficient, a new bottleneck was identified that involves inefficient purification strategies and hence slow-down of structure elucidation of the products of newly discovered gene clusters. Part of the proposed studies in this subproject will focus on the development of efficient methods for purification and structural elucidation of those new compounds. In addition, the genome sequencing efforts of the past grant period identified cases in which the same natural product is made in different organisms by very different biosynthetic routes. We will investigate these examples of convergent evolution. Finally, many natural product phosphonates are produced as short peptides to promote uptake by peptide permeases in the target organisms. Whereas for some compounds small non-ribosomal peptide synthetase (NRPS) clusters are responsible for peptide bond formation, many phosphonate containing peptides are assembled in different, often unidentified ways. We will attempt to provide more insight into this very important aspect with respect to possible future engineering efforts.