Immobilization of a recombinant endo-1,5-arabinanase secreted by Aspergillus nidulans strain A773

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• 作者：André ; Ricardo de Lima Damá ; sio^a ; Benevides Costa Pessela^b ; Cé ; sar Mateo^c ; Fernando Segato^d ; Rolf Alexander Prade^e ; Jose Manuel Guisan^c ; Maria de Lourdes Teixeira de Moraes Polizeli^f ; ^{polizeli@ffclrp.usp.br}
• 关键词：Aspergillus niveus ; Aspergillus nidulans ; Recombinant endo-1 ; 5-arabinanase ; Over-expression ; Purification ; Immobilization
• 刊名：Journal of Molecular Catalysis B ; Enzymatic
• 出版年：2012
• 期刊代码：55_13811177
• 类别：ce
• 出版时间：May, 2012
• 卷：77
• 期：Complete
• 页码：39-45
• 文件大小：623 K

摘要

An endo-1,5-arabinanase (abnA) encoding gene from Aspergillus niveus was identified, cloned and successfully expressed in Aspergillus nidulans strain A773. Based on amino acid sequence comparison, the 34-kDa enzyme could be assigned to CAZy GH family 43. Characterization of purified recombinant endo-1,5-arabinanase (AbnA) revealed that it is active at a wide pH range (pH 4.0-7.0) and an optimum temperature at 70 掳C. The immobilization of the AbnA was performed via covalent binding onto agarose-modified supports: glyoxyl iminodiacetic acid-Ni²⁺, glyoxyl amine, glyoxyl (4%and 10%) and cyanogen bromide activated sepharose. The yield of immobilization was similar on glyoxyl amine and glyoxyl (96%), and higher than glyoxyl iminodiacetic acid-Ni²⁺ (43%) support. The thermal inactivation of these immobilized preparations showed that the stability of the AbnA immobilized on glyoxyl 4 and 10%was improved by 4.0 and 10.3-fold factor at 70 掳C. The half-life of glyoxyl 4%derivative at 60 掳C was >48 h (pH 5), 9 h (pH 7) and 88 min (pH 9). The major hydrolysis product of debranched arabinan or arabinopentaose by glyoxyl agarose-immobilized AbnA was arabinobiose.