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Annals of Microbiology

, Volume 68, Issue 5, pp 261–271 | Cite as

Screening and identification of an Enterobacter ludwigii strain expressing an active β-xylosidase

  • Jiashi Zhang
  • Tangbing Cui
  • Xueqing Li
Original Article
  • 125 Downloads

Abstract

Researchers have expressed increasing interest in the xylanolytic enzymes used in hemicellulose hydrolysis that convert wood and agricultural residues to second-generation biofuels. In our study, 32 isolates showed clear hydrolysis zones on agar plates containing xylan after Congo red staining. Among these isolates, strain LY-62 exhibited the highest β-xylosidase activity (1.29 ± 0.05 U/mL). According to the phylogenetic analysis of the 16S rDNA, strain LY-62 belongs to the Enterobacter genus. Using a combination of electron microscopy, Gram-staining, and conventional physiological and biochemical examinations, the strain LY-62 was identified as Enterobacter ludwigii. The β-xylosidase gene from Enterobacter ludwigii LY-62 was cloned, and the full-length protein was expressed in Escherichia coli as an N-terminal or C-terminal His-tagged fusions protein. Optimal β-xylosidase activity was achieved at pH 7.0 and 40 °C. The Michaelis constant KM values for His-Xyl62 and Xyl62-His were 1.55 and 2.8 mmol/L, respectively. The kcat values for His-Xyl62 and Xyl62-His were 8.51 and 6.94 s−1, respectively. The catalytic efficiencies of His-Xyl62 and Xyl62-His were 5.49 and 2.48 s−1 × mM−1, respectively. Thus, Xyl62 is a functional β-xylosidase, and our study represents the first report of a β-xylosidase from Enterobacter ludwigii.

Keywords

Enterobacter ludwigii Expression GH43 β-xylosidase 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Research involving human participants and/or animals

Not applicable.

Informed consent

Not applicable.

Ethics approval

Not applicable.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018

Authors and Affiliations

  1. 1.School of Bioscience and BioengineeringSouth China University of TechnologyGuangzhouChina

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