Analysis of the carbohydrate-binding-module from Fragaria x ananassa α-L-arabinofuranosidase 1

MARTINEZ, GUSTAVO ADOLFO; SIN, IGNACIO NICOLAS; CIVELLO, PEDRO MARCOS; PERINI MAURO ALEJANDRO

PLANT PHYSIOLOGY AND BIOCHEMISTRY

ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

Lugar: Paris; Año: 2016 vol. 107 p. 96 - 103

0981-9428

a-L-arabinofuranosidases (EC 3.2.1.55) are enzymes involved in the catabolism of several cell-wallpolysaccharides such as pectins and hemicelluloses, catalyzing the hydrolysis of terminal nonreducinga-L-arabinofuranosil residues. Bioinformatic analysis of the aminoacidic sequences of Fragariax ananassa a-L-arabinofuranosidases predict a putative carbohydrate-binding-module of the familyCBM_4_9, associated to a wide range of carbohydrate affinities. In this study, we report the characterizationof the binding affinity profile to different cell wall polysaccharides of the putative CBM of aeLarabinofuranosidase1 from Fragaria x ananassa (CBM-FaARA1). The sequence encoding for the putativeCBM was cloned and expressed in Escherichia coli, and the resultant recombinant protein was purifiedfrom inclusion bodies by a Nickel affinity chromatography under denaturing conditions. The refoldedrecombinant protein was then subjected to binding assays and affinity gel electrophoresis, which indicatedits ability to bind cellulose and also high affinity for homogalacturonans.