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

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

doi:10.1016/j.abb.2005.10.028

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

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

Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

35 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 language	English
Pages (from-to)	73-82
Number of pages	10
Journal	Archives of Biochemistry and Biophysics
Volume	448
Issue number	1-2
DOIs	https://doi.org/10.1016/j.abb.2005.10.028
State	Published - 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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10.1016/j.abb.2005.10.028

Cite this

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title = "Biosynthetic potential of sesquiterpene synthases: Alternative products of tobacco 5-epi-aristolochene synthase",

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.",

keywords = "Biosynthesis, Electrophilic cyclization, Enzyme mechanism, GC-MS, Gas chromatography, Isomerization, Product identification, Sesquiterpene, Terpene cyclase, Terpene synthase",

author = "O'Maille, {Paul E.} and Joe Chappell and Noel, {Joseph P.}",

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{\textquoteright}Maille is an NIH Postdoctoral Research Fellow (GM069056). Joseph P. Noel is an investigator of the Howard Hughes Medical Institute. ",

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N1 - 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.

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N2 - 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.

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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