Fiche publication
Date publication
janvier 2020
Journal
Journal of visualized experiments : JoVE
Auteurs
Membres identifiés du Cancéropôle Est :
Dr TERRYN Christine
Tous les auteurs :
Terryn C, Habrant A, Paës G, Spriet C
Lien Pubmed
Résumé
In lignocellulosic biomass (LB), the activity of enzymes is limited by the appearance of non-specific interactions with lignin during the hydrolysis process, which maintains enzymes far from their substrate. Characterization of these complex interactions is thus a challenge in complex substrates such as LB. The method here measures molecular interactions between fluorophore-tagged molecules and native autofluorescent lignin, to be revealed by Förster resonance energy transfer (FRET). Contrary to FRET measurements in living cells using two exogenous fluorophores, FRET measurements in plants using lignin is not trivial due to its complex autofluorescence. We have developed an original acquisition and analysis pipeline with correlated observation of two complementary properties of fluorescence: fluorescence emission and lifetime. sFLIM (spectral and fluorescent lifetime imaging microscopy) provides the quantification of these interactions with high sensitivity, revealing different interaction levels between biomolecules and lignin.
Référence
J Vis Exp. 2020 Jan 2;(155):
