, Volume 249, Issue 1, pp 59–70 | Cite as

In planta and in silico characterization of five sesquiterpene synthases from Vitis vinifera (cv. Shiraz) berries

  • Bjørn Dueholm
  • Damian P. Drew
  • Crystal Sweetman
  • Henrik T. SimonsenEmail author
Original Article
Part of the following topical collections:
  1. Terpenes and Isoprenoids


Main conclusion

Five Vitis vinifera sesquiterpene synthases were characterized, two was previously uncharacterized, one being a caryophyllene/cubebene synthase and the other a cadinene synthase. Residue differences with other Vitis sesquiterpene synthases are described.

The biochemical composition of grape berries at harvest can have a profound effect on the varietal character of the wine produced. Sesquiterpenes are an important class of volatile compounds produced in grapes that contribute to the flavor and aroma of wine, making the elucidation of their biosynthetic origin an important field of research. Five cDNAs corresponding to sesquiterpene synthase genes (TPSs) were isolated from Shiraz berries and expressed in planta in Nicotiana benthamiana followed by chemical characterization by GC–MS. Three of the TPS cDNAs were isolated from immature berries and two were isolated from ripe Shiraz berries. Two of the investigated enzymes, TPS26 and TPS27, have been previously investigated by expression in E. coli, and the in planta products generally correspond to these previous studies. The enzyme TPS07 differed by eight amino acids (none of which are in the active site) from germacrene B and D synthase isolated from Gewürztraminer grapes and characterized in vitro. Here in planta characterization of VvShirazTPS07 yielded ylangene, germacrene D and several minor products. Two of the enzymes isolated from immature berries were previously uncharacterized enzymes. VvShirazTPS-Y1 produced cadinene as a major product and at least 17 minor sesquiterpenoid skeletons. The second, VvShirazTPS-Y2, was characterized as a caryophyllene/cubebene synthase, a combination of products not previously reported from a single enzyme. Using in silico methods, we identified residues that could play key roles regarding differences in product formation of these enzymes. The first ring closure that is either a 1,10- or 1,11-ring closure is likely controlled by three neighboring amino acids in helices G1, H2, and J. As for many other investigated TPS enzymes, we also observe that only a few residues can account for radical changes in product formation.


Sesquiterpene synthase Shiraz Isoprenoid Sesquiterpenes Wine aroma 



This research was supported by the European Union Seventh Framework Programme (FP7/2007-2013) under Grant agreement 275422, which supported a Marie Curie International Outgoing Fellowship (DPD). The Danish Strategic Research Council Grant “SPOTLight” supported the work of HTS.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

425_2018_2986_MOESM1_ESM.pdf (2 mb)
Online Resource 1: Product profiles of the five characterized VvShirazTPS enzymes (PDF 2072 kb)
425_2018_2986_MOESM2_ESM.pdf (167 kb)
Online Resource 2: Full-length sequence alignment of the VvShirazTPS enzymes (PDF 166 kb)
425_2018_2986_MOESM3_ESM.pdf (196 kb)
Online Resource 3: Positions of residues Ser, Ser, and Met in VvShirazTPS27 corresponding to residue-locations 402, 440, and 516 in TEAS. Distances from the residues to C10 of FPP are shown (PDF 195 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biotechnology and BiomedicineTechnical University of DenmarkLyngbyDenmark
  2. 2.Wine Science, School of Agriculture, Food and WineUniversity of AdelaideUrrbraeAustralia

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