Combined mass mapping and biochemical characterization of grape beta-glycosidase-enriched extract.
A beta-glucosidase enzyme activity was enriched from skins of ripe grape berry by cell wall fractionation, hydrophobic interaction and cation-exchange chromatographies. This enriched enzyme extract contained several beta-glycosidase activities hydrolyzing a wide range of synthetic and natural monoglycosides and diglycosides, as well as a beta-fructosidase activity. The enzyme extract was further characterized by two-dimensional gel electrophoresis coupled to peptide mass fingerprinting of eight spots using MALDI-TOF mass spectrometry. No beta-glucosidase but a beta-fructosidase associated to the relevant spot at 66 kDa/pI 5.1 was identified. Taken together all results issued from the biochemical characterization, the substrate specificity and the mass spectrometry-based identification of this enriched enzyme extract, we propose that this protein could be a specific beta-fructosidase isoform associated with a broad spectrum of beta-glycosidase activities in grape berry skin and involved in cell wall modifications which occur during the ripening-induced thickness of the grape.
Sarry
Jean-Emmanuel
JE
UMR UMI/ENSAM/INRA Science Pour l'Oenologie, INRA, 2 Place Pierre Viala, F-34060, Montpellier Cedex 1, France. jesarry@sas.upenn.edu
Grimplet
Jérôme
J
Sommerer
Nicolas
N
Vallier
Marie-José
MJ
Pradal
Martine
M
Mondolot
Laurence
L
Andary
Claude
C
Günata
Ziya
Z
Romieu
Charles
C
D002473:Cell Wall / Q000201:enzymology; D015180:Electrophoresis, Gel, Two-Dimensional; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D027843:Vitis / Q000201:enzymology*; D043324:beta-Fructofuranosidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D043324:beta-Fructofuranosidase