海洋微生物次级代谢产物往往具有独特、新颖和复杂的化学结构，为发掘新颖酶学机理提供了良好的材料。从海洋异壁放线菌Actinoalloteichus cyanogriseus WH1-2216-6 中分离鉴定到新生物碱类化合物cyanogramide，该化合物具有独特的螺环的吡咯并[1,2-c]咪唑结构，可有效的降低肿瘤细胞对阿霉素（adriamycin）和长春新碱（vincristine）产生的抗性。为在阐明生物合成机制的基础上获得更多结构衍生物，拟开展以下内容的研究：（1）克隆鉴定cyanogramide生物合成基因簇；（2）获得高产cyanogramide的工程菌株；（3）解析cyanogramide结构中螺环的吡咯并[1,2-c]咪唑结构生物合成机制，确定关键酶的催化机理；（4）在解析生物合成途径的基础上，利用组合生物合成技术进行结构衍生和多样化，初步开展构效关系研究。

The secondary metabolites produced by marine microorganisms often have unique, novel and complex chemical structure, which provided materials for exploring novel enzymatic mechanisms. Cyanogramide, an unprecedented alkaloid bearing a novel spirocyclic pyrrolo[1,2-c]imidazole skeleton, was isolated from the marine-derived Actinoalloteichus cyanogriseus WH1-2216-6.Cyanogramide could efficiently reverse the adriamycin-induced resistance of K562/A02 and MCF-7/Adr cells, and the vincristine-induced resistance of KB/VCR cells.The project intends to carry out the following studies:(1) cloning and identification the biosynthesis gene cluster of cyanogramide; (2)constructing the engineering strain for high producing cyanogramide; （3）elucidating tailoring enzymes in cyanogramide biosynthesis for spirocyclic pyrrolo[1,2-c]imidazole skeleton; (4) generating cyanogramide analogues by combinatorial biosynthesis methods for studying their structure-activity relationships.