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

SIN, IN; PERINI, MA; MARTÍNEZ, GA; CIVELLO, PM

PLANT PHYSIOLOGY AND BIOCHEMISTRY

ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

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

0981-9428

α-L-arabinofuranosidases (EC 3.2.1.55) are enzymes involved in thecatabolism of several cell-wall polysaccharides such as pectins andhemicelluloses, catalyzing the hydrolysis of terminal non-reducing α-L-arabinofuranosilresidues. Bioinformatic analysis on the aminoacidic sequence of Fragaria x ananassa α-L-arabinofuranosidasespredict a putative carbohydrate-binding-module of the family CBM_4_9,associated to a wide range of carbohydrate affinities. In this study, we reportthe characterization of the binding affinity profile to different cell wallpolysaccharides of the putative CBM of α?L-arabinofuranosidase 1 from Fragaria x ananassa (CBM-FaARA1). Thesequence encoding for the putative CBM was cloned and expressed in E. coli, and the resultant recombinantprotein was purified from inclusion bodies by a Nickel affinity chromatographyunder denaturing conditions. The refolded recombinant protein was thensubjected to binding assays and Affinity Gel Electrophoresis, which indicated itsability to bind cellulose and also high affinity to homogalacturonans.