Formation of a Novel Macrocyclic Alkaloid from the Unnatural Farnesyl Diphosphate Analogue Anilinogeranyl Diphosphate by 5‑Epi-Aristolochene Synthase

• Published in: ACS chemical biology Vol. 10; no. 7; pp. 1729 - 1736
• Main Authors: Rising, Kathleen A, Crenshaw, Charisse M, Koo, Hyun Jo, Subramanian, Thangaiah, Chehade, Kareem A. H, Starks, Courtney, Allen, Keith D, Andres, Douglas A, Spielmann, H. Peter, Noel, Joseph P, Chappell, Joe
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 17-07-2015
• Subjects: Alkaloids - chemistry; Alkaloids - metabolism; Alkyl and Aryl Transferases - chemistry; Alkyl and Aryl Transferases - metabolism; Macrocyclic Compounds - chemistry; Macrocyclic Compounds - metabolism; Models, Molecular; Nicotiana - chemistry; Nicotiana - enzymology; Nicotiana - metabolism; Polyisoprenyl Phosphates - chemistry; Polyisoprenyl Phosphates - metabolism; Sesquiterpenes - chemistry; Sesquiterpenes - metabolism; Terpenes - chemistry; Terpenes - metabolism
• ISSN: 1554-8929; 1554-8937
• DOI: 10.1021/acschembio.5b00145

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.