Biosynthetic potential of sesquiterpene synthases: Alternative products of tobacco 5-epi-aristolochene synthase

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• 作者：Paul E. O’ ; Maille ; Joe Chappell and Joseph P. Noel
• 关键词：Sesquiterpene ; Terpene synthase ; Terpene cyclase ; GC– ; MS ; Gas chromatography ; Biosynthesis ; Product identification ; Isomerization ; Enzyme mechanism ; Electrophilic cyclization
• 刊名：Archives of Biochemistry and Biophysics
• 出版年：2006
• 出版时间：15 April 2006
• 年：2006
• 卷：448
• 期：1-2
• 页码：73-82
• 全文大小：489 K

文摘

Nicotiana tabacum (tobacco) 5-epi-aristolochene synthase (TEAS) serves as an useful model for understanding the enzyme mechanisms of sesquiterpene biosynthesis. Despite extensive bio-chemical and structural characterization of TEAS, a more detailed analysis of the reaction product spectrum is lacking. This study reports the discovery and quantification of several alternative sesquiterpene products generated by recombinant TEAS in the single-vial GC–MS assay. The combined use of chiral and non-polar stationary phases for gas chromatography separations proved critical for resolving the numerous sesquiterpene products of TEAS for mass spectral analysis and identification. Co-injection studies with available authentic standards from both synthetic and natural sources further corroborated the assignment of several compounds, resulting in an annotated reaction mechanism accounting for their biosynthesis. Moreover, a previously undocumented farnesyl trans–cis isomerization pathway was observed.