Plant Molecular Biology

, Volume 63, Issue 1, pp 49–61 | Cite as

Glucosinolate hydrolysis in Lepidium sativum––identification of the thiocyanate-forming protein

  • Meike Burow
  • Andrea Bergner
  • Jonathan Gershenzon
  • Ute WittstockEmail author


Glucosinolates are a class of thioglycosides found predominantly in plants of the order Brassicales whose function in anti-herbivore defense has been attributed to the products formed by myrosinase-catalyzed hydrolysis upon plant tissue damage. The most common type of hydrolysis products, the isothiocyanates, are toxic to a wide range of organisms. Depending on the glucosinolate side-chain structure and the presence of certain protein factors, other types of hydrolysis products, such as simple nitriles, epithionitriles and organic thiocyanates, can be formed whose biological functions are not well understood. Of the proteins controlling glucosinolate hydrolysis, only epithiospecifier proteins (ESPs) that promote the formation of simple nitriles and epithionitriles have been identified on a molecular level. We investigated glucosinolate hydrolysis in Lepidium sativum and identified a thiocyanate-forming protein (TFP) that shares 63-68% amino acid sequence identity with known ESPs and up to 55% identity with myrosinase-binding proteins from Arabidopsis thaliana, but differs from ESPs in its biochemistry. TFP does not only catalyze thiocyanate and simple nitrile formation from benzylglucosinolate but also the formation of simple nitriles and epithionitriles from aliphatic glucosinolates. Analyses of glucosinolate hydrolysis products in L. sativum autolysates and TFP transcript accumulation revealed an organ-specific regulation of thiocyanate formation. The identification of TFP defines a new family of proteins that control glucosinolate hydrolysis and challenges the previously proposed reaction mechanism of epithionitrile formation. As a protein that promotes the formation of a wide variety of hydrolysis products, its identification provides an important tool for further elucidating the mechanisms of glucosinolate hydrolysis as well as the ecological role and the evolutionary origin of the glucosinolate-myrosinase system.


Epithionitrile Epithiospecifier protein Glucosinolate hydrolysis Lepidium sativum Myrosinase-binding protein Organic thiocyanate 



Epithiospecifier protein


Thiocyanate-forming protein



We thank Michael Reichelt for providing intact glucosinolates and the Max Planck Society for financial support.


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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Meike Burow
    • 1
    • 2
  • Andrea Bergner
    • 1
  • Jonathan Gershenzon
    • 1
  • Ute Wittstock
    • 1
    • 2
    Email author
  1. 1.Department of BiochemistryMax Planck Institute for Chemical EcologyJenaGermany
  2. 2.Institut für Pharmazeutische BiologieTechnische Universität BraunschweigBraunschweigGermany

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