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Applied Microbiology and Biotechnology

, Volume 103, Issue 9, pp 3783–3793 | Cite as

Highly thermostable GH51 α-arabinofuranosidase from Hungateiclostridium clariflavum DSM 19732

  • Alei GengEmail author
  • Jian Wu
  • Rongrong Xie
  • Hongcheng Wang
  • Yanfang Wu
  • Xia Li
  • Fuxiang Chang
  • Jianzhong SunEmail author
Biotechnologically relevant enzymes and proteins
  • 119 Downloads

Abstract

Arabinofuranosidase plays an essential role in the process of hydrolysis of arabinoxylan (AX). Thermostable, versatile, and efficient arabinofuranosidase is thus of great interest for the biorefinery industry. A GH51 arabinofuranosidase, Abf51, from Hungateiclostridium clariflavum DSM 19732 was heterogeneously expressed in Escherichia coli. Abf51 was found to have an optimal pH and temperature of 6.5 and 60 °C, respectively, with very high thermostability. At the optimal working temperature (60 °C), Abf51 retained over 90% activity after a 2-day incubation and over 60% activity after a 6-day incubation. Abf51 could effectively remove the arabinofuranosyls from three kinds of AX oligosaccharides [23-α-l-arabinofuranosyl-xylotriose (A2XX), 32-α-l-arabinofuranosyl-xylobiose (A3X), and 2333-di-α-l-arabinofuranosyl-xylotriose (A2 + 3XX)], which characterized as either single substitution or double substitution by arabinofuranosyls on terminal xylopyranosyl units. The maximal catalytic efficiency (Kcat/Km) was observed using p-nitrophenyl-α-l-arabinofuranoside (pNPAF) as a substrate (205.0 s−1 mM−1), followed by using A3X (22.8 s−1 mM−1), A2XX (6.9 s−1 mM−1), and A2 + 3XX (0.5 s−1 mM−1) as substrates. Moreover, the presence of Abf51 significantly stimulated the saccharification level of AX (18.5 g L−1) up to six times along with a β-xylanase as well as a β-xylosidase. Interestingly, in our survey of top thermostable arabinofuranosidases, most members were found from GH51, probably due to their owning of (β/α)8-barrel architectures. Our results suggested the great importance of GH51s as candidates for thermostable, versatile, and efficient arabinofuranosidases toward industry application.

Keywords

Arabinofuranosidase Thermostable Hungateiclostridium clariflavum Arabinoxylan 

Notes

Funding

This work was financially supported by the Postdoctoral Science Foundation of China (2018T110447), Jiangsu Key Laboratory for Biomass Energy and Material (JSBEM201807), National Natural Science Foundation of China (31772529), and the Startup Foundation of Jiangsu University (11JDG109). Hongcheng Wang acknowledges the supports of the Natural Science Foundation of Jiangsu Province (BK20170541) and the Priority Academic Program Development of Jiangsu Higher Education Institution.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9753_MOESM1_ESM.pdf (319 kb)
ESM 1 (PDF 319 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Biofuels institute, School of the Environment and Safety EngineeringJiangsu UniversityZhenjiangChina
  2. 2.Jiangsu Key Laboratory for Biomass Energy and MaterialNanjingChina

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