Cloning and Characterization of a Novel Cold-active Bifunctional Xylosidase/Arabinfuranosidease AX543

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.025

Abstract

Abstract: This work aims to obtain the cold-active xylosidase with high catalytic activity at low temperature,and to study its heterologous expression,as well as analyze its enzymatic characteristics. A novel psychrophilic bifunctional xylosidase/arabinofuranosidase gene ax543 in GH43 family was cloned using Touch down PCR and TAIL PCR,and was successfully expressed in Pichia pastoris. Analysis of biochemical characterization suggested that recombinant enzyme AX543 exhibited optimal activity at 25℃,and remained 54% and 21% relative activities at 15℃ and 4℃. Moreover,AX543 with xylosidase and arabinofuranosidase activity hydrolyzed xylobiose,xylotriose,birchwood xylan,beechwood xylan,and wheat arabinoxylan,and showed a 1.46 fold synergic effect with xylanase Xyn11-1. AX543 also presented xylose and arabinose tolerance with K_i 84.78 mmol/L and 54.01 mmol/L.

Key words: xylosidase, arabinofuranosidase, bifunctional, cold-active, synergic effect, product tolerance

ZHANG Ming-hui, LI Zhong-yuan, QIU Hai-yan, WANG Cui-qiong, WANG Hui, ZHANG Tong-cun. Cloning and Characterization of a Novel Cold-active Bifunctional Xylosidase/Arabinfuranosidease AX543[J]. Biotechnology Bulletin, 2016, 32(11): 215-223.

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