Computational-guided discovery and characterization of a sesquiterpene synthase from Streptomyces clavuligerus

Jeng Yeong Chow, Bo Xue Tian, Gurusankar Ramamoorthy, Brandan S. Hillerich, Ronald D. Seidel, Steven C. Almo, Matthew P. Jacobson, C. Dale Poulter

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Terpenoids are a large structurally diverse group of natural products with an array of functions in their hosts. The large amount of genomic information from recent sequencing efforts provides opportunities and challenges for the functional assignment of terpene synthases that construct the carbon skeletons of these compounds. Inferring function from the sequence and/or structure of these enzymes is not trivial because of the large number of possible reaction channels and products. We tackle this problem by developing an algorithm to enumerate possible carbocations derived from the farnesyl cation, the first reactive intermediate of the substrate, and evaluating their steric and electrostatic compatibility with the active site. The homology model of a putative pentalenene synthase (Uniprot: B5GLM7) from Streptomyces clavuligerus was used in an automated computational workflow for product prediction. Surprisingly, the workflow predicted a linear triquinane scaffold as the top product skeleton for B5GLM7. Biochemical characterization of B5GLM7 reveals the major product as (5S,7S,10R,11S)-cucumene, a sesquiterpene with a linear triquinane scaffold. To our knowledge, this is the first documentation of a terpene synthase involved in the synthesis of a linear triquinane. The success of our prediction for B5GLM7 suggests that this approach can be used to facilitate the functional assignment of novel terpene synthases.

Original languageEnglish (US)
Pages (from-to)5661-5666
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number18
DOIs
StatePublished - May 5 2015

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Sesquiterpenes
Streptomyces
Workflow
Skeleton
Terpenes
Biological Products
Static Electricity
Documentation
Cations
Catalytic Domain
Carbon
Enzymes
terpene synthase

Keywords

  • Annotation
  • Cyclase
  • Sesquiterpene
  • Structure-function

ASJC Scopus subject areas

  • General

Cite this

Computational-guided discovery and characterization of a sesquiterpene synthase from Streptomyces clavuligerus. / Chow, Jeng Yeong; Tian, Bo Xue; Ramamoorthy, Gurusankar; Hillerich, Brandan S.; Seidel, Ronald D.; Almo, Steven C.; Jacobson, Matthew P.; Poulter, C. Dale.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, No. 18, 05.05.2015, p. 5661-5666.

Research output: Contribution to journalArticle

Chow, Jeng Yeong ; Tian, Bo Xue ; Ramamoorthy, Gurusankar ; Hillerich, Brandan S. ; Seidel, Ronald D. ; Almo, Steven C. ; Jacobson, Matthew P. ; Poulter, C. Dale. / Computational-guided discovery and characterization of a sesquiterpene synthase from Streptomyces clavuligerus. In: Proceedings of the National Academy of Sciences of the United States of America. 2015 ; Vol. 112, No. 18. pp. 5661-5666.
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