Improvement of fungal arabinofuranosidase thermal stability by reversible immobilization

A gene encoding a-L-arabinofuranosidase (abfA) from Aspergillus niveus was identified, cloned, and successfully expressed in Aspergillus nidulans. Based on amino acid sequence comparison, the 88.6 kDa enzyme could be assigned to the GH family 51. The characterization of the purified recombinant AbfA revealed that the enzyme was active at a limited pH range (pH 4.0-5.0) and an optimum temperature of 70 degrees C. The AbfA was able to hydrolyze arabinoxylan, xylan from birchwood, debranched arabinan, and 4-nitrophenyl arabinofuranoside. Synergistic reactions using both AbfA and endoxylanase were also assessed. The highest degree of synergy was obtained after the sequential treatment of the substrate with endoxylanase, followed by AbfA, which was observed to release noticeably more reducing sugars than that of either enzyme acting individually. The immobilization of AbfA was performed via ionic adsorption onto various supports: agarose activated by polyethyleneimine polymers, cyanogen bromide activated Sepharose, DEAE-Sepharose, and Sepharose-Q The Sepharose-Q derivative remained fully active at pH 5 after 360 min at 60 degrees C, whereas the free AbfA was inactivated after 60 min. A synergistic effect of arabinoxylan hydrolysis by AbfA immobilized in Sepharose-Q and endoxylanase immobilized in glyoxyl agarose was also observed. The stabilization of arabinofuranosidases using immobilization tools is a novel and interesting topic. (C) 2012 Elsevier Ltd. All rights reserved.National System for Research on Biodiversity (SisbiotaBrazil Conselho de Desenvolvimento Cientifico e Tecnologico, CNPq)National System for Research on Biodiversity (Sisbiota-Brazil - Conselho de Desenvolvimento Cientifico e Tecnologico, CNPq) [563260/2010-6]National System for Research on Biodiversity (Sisbiota-Brazil - Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, FAPESP) [2010/52322-3]National System for Research on Biodiversity (SisbiotaBrazil Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, FAPESP)Banco SANTANDER S.A.Banco SANTANDER S.A.FAPESPFAPESPDepartment of Energy [06103-OKL, ZDJ-7-77608-01]Department of EnergyMinisterio de Ciencia e Innovacion, Spain [AGL-2009-07625]Ministerio de Ciencia e Innovacion, Spai