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Title: Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution

Abstract

The mint family (Lamiaceae) is well documented as a rich source of terpene natural products. More than 200 diterpene skeletons have been reported from mints, but biosynthetic pathways are known for just a few of these. We crossreferenced chemotaxonomic data with publicly available transcriptomes to select common selfheal (Prunella vulgaris) and its highly unusual vulgarisin diterpenoids as a case study for exploring the origins of diterpene skeletal diversity in Lamiaceae. Four terpene synthases (TPS) from the TPS-a subfamily, including two localised to the plastid, were cloned and functionally characterised. Previous examples of TPS-a enzymes from Lamiaceae were cytosolic and reported to act on the 15-carbon farnesyl diphosphate. Plastidial TPS-a enzymes using the 20-carbon geranylgeranyl diphosphate are known from other plant families, having apparently arisen independently in each family. All four new enzymes were found to be active on multiple prenyl-diphosphate substrates with different chain lengths and stereochemistries. One of the new enzymes catalysed the cyclisation of geranylgeranyl diphosphate into 11-hydroxy vulgarisane, the likely biosynthetic precursor of the vulgarisins. We uncovered the pathway to a rare diterpene skeleton. Our results support an emerging paradigm of substrate and compartment switching as important aspects of TPS evolution and diversification.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [1];  [1];  [1]; ORCiD logo [1]
  1. Michigan State Univ., East Lansing, MI (United States). Dept. of of Biochemistry and Molecular Biology
  2. Michigan State Univ., East Lansing, MI (United States). Dept. of of Biochemistry and Molecular Biology and Dept. of Pharmacology and Toxicology
Publication Date:
Research Org.:
Univ. of Wisconsin, Madison, WI (United States). Great Lakes Bioenergy Research Center
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)
OSTI Identifier:
1546989
Grant/Contract Number:  
FC02-07ER64494; SC0018409; 1737898.
Resource Type:
Accepted Manuscript
Journal Name:
New Phytologist
Additional Journal Information:
Journal Volume: 223; Journal Issue: 1; Journal ID: ISSN 0028-646X
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Diterpenoid; Prunella vulgaris (common selfheal); terpene synthase; transcriptome; transit peptide; vulgarisane

Citation Formats

Johnson, Sean R., Bhat, Wajid Waheed, Sadre, Radin, Miller, Garret P., Garcia, Alekzander Sky, and Hamberger, Björn. Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution. United States: N. p., 2019. Web. doi:10.1111/nph.15778.
Johnson, Sean R., Bhat, Wajid Waheed, Sadre, Radin, Miller, Garret P., Garcia, Alekzander Sky, & Hamberger, Björn. Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution. United States. doi:10.1111/nph.15778.
Johnson, Sean R., Bhat, Wajid Waheed, Sadre, Radin, Miller, Garret P., Garcia, Alekzander Sky, and Hamberger, Björn. Mon . "Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution". United States. doi:10.1111/nph.15778. https://www.osti.gov/servlets/purl/1546989.
@article{osti_1546989,
title = {Promiscuous terpene synthases from Prunella vulgaris highlight the importance of substrate and compartment switching in terpene synthase evolution},
author = {Johnson, Sean R. and Bhat, Wajid Waheed and Sadre, Radin and Miller, Garret P. and Garcia, Alekzander Sky and Hamberger, Björn},
abstractNote = {The mint family (Lamiaceae) is well documented as a rich source of terpene natural products. More than 200 diterpene skeletons have been reported from mints, but biosynthetic pathways are known for just a few of these. We crossreferenced chemotaxonomic data with publicly available transcriptomes to select common selfheal (Prunella vulgaris) and its highly unusual vulgarisin diterpenoids as a case study for exploring the origins of diterpene skeletal diversity in Lamiaceae. Four terpene synthases (TPS) from the TPS-a subfamily, including two localised to the plastid, were cloned and functionally characterised. Previous examples of TPS-a enzymes from Lamiaceae were cytosolic and reported to act on the 15-carbon farnesyl diphosphate. Plastidial TPS-a enzymes using the 20-carbon geranylgeranyl diphosphate are known from other plant families, having apparently arisen independently in each family. All four new enzymes were found to be active on multiple prenyl-diphosphate substrates with different chain lengths and stereochemistries. One of the new enzymes catalysed the cyclisation of geranylgeranyl diphosphate into 11-hydroxy vulgarisane, the likely biosynthetic precursor of the vulgarisins. We uncovered the pathway to a rare diterpene skeleton. Our results support an emerging paradigm of substrate and compartment switching as important aspects of TPS evolution and diversification.},
doi = {10.1111/nph.15778},
journal = {New Phytologist},
number = 1,
volume = 223,
place = {United States},
year = {2019},
month = {4}
}

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