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

Producción científica: Article › revisión exhaustiva

39 Citas (Scopus)

Resumen

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.

Idioma original	English
Páginas (desde-hasta)	73-82
Número de páginas	10
Publicación	Archives of Biochemistry and Biophysics
Volumen	448
N.º	1-2
DOI	https://doi.org/10.1016/j.abb.2005.10.028
Estado	Published - abr 15 2006

Nota bibliográfica

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.

Financiación

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.

Financiadores	Número del financiador
National Institutes of Health (NIH)
National Institute of General Medical Sciences	R01GM054029

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/j.abb.2005.10.028

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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",

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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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KW - Sesquiterpene

KW - Terpene cyclase

KW - Terpene synthase

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

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

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