Advanced search
1 file | 736.86 KB

Shikimate hydroxycinnamoyl transferase (HCT) activity assays in Populus nigra

Igor Cesarino (UGent) , Ruben Vanholme (UGent) , Geert Goeminne (UGent) , Bartel Vanholme (UGent) and Wout Boerjan (UGent)
(2013) BIO-PROTOCOL. 3(22).
Author
Organization
Project
Biotechnology for a sustainable economy (Bio-Economy)
Abstract
Lignin is a complex phenolic polymer deposited in secondarily-thickened plant cell walls. The polymer is mainly derived from the three primary monolignols: p-coumaryl, coniferyl and sinapyl alcohol which give rise to p-hydroxyphenyl, guaiacyl and syringyl units (H, G and S units, respectively) when coupled into the polymer. The building blocks differ in their degree of methoxylation and their biosynthetic pathway is catalyzed by more than 10 enzymes. HCT plays a crucial role by channeling the phenylpropanoids towards the production of coniferyl and sinapyl alcohols. Interestingly, HCT has been reported to be implicated in the pathway both upstream and downstream of the 3-hydroxylation of the aromatic ring of p-coumaroyl shikimate (Figure 1) (Hoffmann et al., 2003; Hoffmann et al., 2004; Vanholme et al., 2013b). These features highlight the importance of developing an assay to reliably measure HCT activity in planta. Here, we describe a UPLC-MS-based method for the analysis of HCT activity in xylem total protein extracts of Populus nigra, which can be adapted to other woody and herbaceous plant species. The protocol was initially described in Vanholme et al. (2013a).

Downloads

  • Cesarino et al. 2013 Bio-protocol 3 e978.pdf
    • full text
    • |
    • open access
    • |
    • PDF
    • |
    • 736.86 KB

Citation

Please use this url to cite or link to this publication:

Chicago
Cesarino, Igor, Ruben Vanholme, Geert Goeminne, Bartel Vanholme, and Wout Boerjan. 2013. “Shikimate Hydroxycinnamoyl Transferase (HCT) Activity Assays in Populus Nigra.” Bio-protocol 3 (22).
APA
Cesarino, I., Vanholme, R., Goeminne, G., Vanholme, B., & Boerjan, W. (2013). Shikimate hydroxycinnamoyl transferase (HCT) activity assays in Populus nigra. BIO-PROTOCOL, 3(22).
Vancouver
1.
Cesarino I, Vanholme R, Goeminne G, Vanholme B, Boerjan W. Shikimate hydroxycinnamoyl transferase (HCT) activity assays in Populus nigra. BIO-PROTOCOL. 2013;3(22).
MLA
Cesarino, Igor, Ruben Vanholme, Geert Goeminne, et al. “Shikimate Hydroxycinnamoyl Transferase (HCT) Activity Assays in Populus Nigra.” BIO-PROTOCOL 3.22 (2013): n. pag. Print.
@article{5772977,
  abstract     = {Lignin is a complex phenolic polymer deposited in secondarily-thickened plant cell walls. The polymer is mainly derived from the three primary monolignols: p-coumaryl, coniferyl and sinapyl alcohol which give rise to p-hydroxyphenyl, guaiacyl and syringyl units (H, G and S units, respectively) when coupled into the polymer. The building blocks differ in their degree of methoxylation and their biosynthetic pathway is catalyzed by more than 10 enzymes. HCT plays a crucial role by channeling the phenylpropanoids towards the production of coniferyl and sinapyl alcohols. Interestingly, HCT has been reported to be implicated in the pathway both upstream and downstream of the 3-hydroxylation of the aromatic ring of p-coumaroyl shikimate (Figure 1) (Hoffmann et al., 2003; Hoffmann et al., 2004; Vanholme et al., 2013b). These features highlight the importance of developing an assay to reliably measure HCT activity in planta. Here, we describe a UPLC-MS-based method for the analysis of HCT activity in xylem total protein extracts of Populus nigra, which can be adapted to other woody and herbaceous plant species. The protocol was initially described in Vanholme et al. (2013a).},
  articleno    = {e978},
  author       = {Cesarino, Igor and Vanholme, Ruben and Goeminne, Geert and Vanholme, Bartel and Boerjan, Wout},
  issn         = {2331-8325},
  journal      = {BIO-PROTOCOL},
  language     = {eng},
  number       = {22},
  pages        = {8},
  title        = {Shikimate hydroxycinnamoyl transferase (HCT) activity assays in Populus nigra},
  url          = {http://www.bio-protocol.org/e978},
  volume       = {3},
  year         = {2013},
}