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

Paul E. O'Maille, Joe Chappell, Joseph P. Noel

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)73-82
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume448
Issue number1-2
DOIs
StatePublished - Apr 15 2006

Bibliographical note

Funding Information:
We thank T. Kollner for the celery seed extract, J.W. Mansfield for the LTC1 clone, and R. Coates for synthetic standards of (−)-4-epi-eremophilene and (−)-prezizaene. We appreciate the insightful comments of Robert Coates, Bryan Greenhagen, and Andy Hess on various aspects of the mechanistic work. We are grateful to the National Institutes of Health for a grant that supported this work (GM54029 to Joseph P. Noel/Joseph Chappell). Paul E. O’Maille is an NIH Postdoctoral Research Fellow (GM069056). Joseph P. Noel is an investigator of the Howard Hughes Medical Institute.

Keywords

  • Biosynthesis
  • Electrophilic cyclization
  • Enzyme mechanism
  • GC-MS
  • Gas chromatography
  • Isomerization
  • Product identification
  • Sesquiterpene
  • Terpene cyclase
  • Terpene synthase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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