Formation of a Novel Macrocyclic Alkaloid from the Unnatural Farnesyl Diphosphate Analogue Anilinogeranyl Diphosphate by 5-Epi-Aristolochene Synthase.
Formation of a Novel Macrocyclic Alkaloid from the Unnatural Farnesyl Diphosphate Analogue Anilinogeranyl Diphosphate by 5-Epi-Aristolochene Synthase.
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DOI：
10.1021/acschembio.5b00145
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发表时间：
2015-07-17
影响因子：
4
通讯作者：
Chappell J
中科院分区：
文献类型：
--
作者：
Rising KA;Crenshaw CM;Koo HJ;Subramanian T;Chehade KA;Starks C;Allen KD;Andres DA;Spielmann HP;Noel JP;Chappell J
作为寻找适合于解析萜类合成酶重要结构特征的底物类似物的一部分，研究了烟草5-表马兜铃烯合成酶与苯丙二磷酸(AGPP)孵育对5-表马兜铃烯生物合成的抑制作用。这种抑制的明显非竞争性性质支持进一步评估AGPP可能如何与酶的结晶形式结合。令人惊讶的是，抑制剂的结合形式似乎经历了与FPP催化相关的天然机制一致的环化事件。通过高分辨GC-MS和核磁共振分析，证实了一种新的13元大环对环烷生物碱的生物催化形成。这项工作为产生新的、功能多样化的中等大小的萜类生物碱提供了新的生物合成方法。
As part of an effort to identify substrate analogs suitable for helping to resolve structural features important for terpene synthases, the inhibition of 5-epi-aristolochene biosynthesis from farnesyl diphosphate (FPP) by the tobacco 5-epi-aristolochene synthase incubated with anilinogeranyl diphosphate (AGPP) was examined. The apparent noncompetitive nature of the inhibition supported further assessment of how AGPP might be bound to crystallographic forms of the enzyme. Surprisingly, the bound form of the inhibitor appeared to have undergone a cyclization event consistent with the native mechanism associated with FPP catalysis. Biocatalytic formation of a novel 13-membered macrocyclic paracyclophane alkaloid was confirmed by high-resolution GC-MS and NMR analysis. This work provides insights into new biosynthetic means for generating novel, functionally diversified, medium-sized terpene alkaloids.