磷酸酰胺天然产物是生物体产生的含有氮磷键的化合物，因为其结构与磷酸化的生物小分子相似，所以容易被细胞内某些酶错误的识别、结合，但是不能被有效的催化。磷酸酰胺天然产物通过抑制靶标生物生命活动某些必须酶的功能，表现出各种各样的生物活性，比如菜豆菌毒素（Phaseolotoxin，PHT）就是一种广谱的生物除草剂。迄今为止发现的磷酸酰胺天然产物仅14个，对于磷酸酰胺天然产物氮磷键的生物合成机制认识也非常有限。PHT含有最多的氮磷键数目(三根)，本项目计划以PHT为模型，研究氮磷键的生物合成机制。我们计划通过基因中断和合成途径重构，鉴定中间体，确定PHT氮磷键合成直接相关的基因。然后通过体外生化实验解析PHT氮磷键的生物合成过程，辅以酶学和结构生物学手段揭示关键反应的催化机制。在对氮磷键的生物合成形成规律性认识的基础上，我们将开展基因组挖掘和组合生物合成，以便发现和创造更多的磷酸酰胺天然产物。

Phosphoramidate natural products refer to a group of Nitrogen-Phosphorus bond (N-P bond) containing compounds produced by living organisms. Attribute to the structure similarity with phosphate biomolecules, it was liable to be misrecognized and bound by certain enzymes in vivo, while could not be catalyzed successfully. Phosphoramidate natural products show diverse bioactivity by inhibiting some essential enzymes of the target organisms, such as Phaseolotoxin is a broad-spectrum biopesticide. Thus far, only fourteen phosphoramidate natural product molecules have been discovered and our understanding about its N-P bond biosynthesis is really limited. Phaseolotoxin, which contains the most N-P bonds number, was selected as a model for N-P bond biosynthesis study in this project. Gene interruption, biosynthetic pathway rebuilding, and biosynthetic intermediates characterization would be employed to identify genes directly related to N-P bond formation. Then, in vitro biochemical experiments would be conducted to address N-P bond biosynthetic pathway. Key reaction mechanisms would be deciphered by enzymology and structural biology experiments. Based on the general understanding of N-P bond formation mechanism, genome mining and combinatorial biosynthesis of phosphoramidate natural products would be carried out as well.