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A new acidophilic endo-β-1,4-xylanase from Penicillium oxalicum: cloning, purification, and insights into the influence of metal ions on xylanase activity

  • Bioenergy/Biofuels/Biochemicals
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A new acidophilic xylanase (XYN11A) from Penicillium oxalicum GZ-2 has been purified, identified and characterized. Synchronized fluorescence spectroscopy was used for the first time to evaluate the influence of metal ions on xylanase activity. The purified enzyme was identified by MALDI TOF/TOF mass spectrometry, and its gene (xyn11A) was identified as an open reading frame of 706 bp with a 68 bp intron. This gene encodes a mature protein of 196 residues with a predicted molecular weight of 21.3 kDa that has the 100 % identity with the putative xylanase from the P. oxalicum 114-2. The enzyme shows a structure comprising a catalytic module family 10 (GH10) and no carbohydrate-binding module family. The specific activities were 150.2, 60.2, and 72.6 U/mg for beechwood xylan, birchwood xylan, and oat spelt xylan, respectively. XYN11A exhibited optimal activity at pH 4.0 and remarkable pH stability under extremely acidic condition (pH 3). The specific activity, K m and V max values were 150.2 U/mg, 30.7 mg/mL, and 403.9 μmol/min/mg for beechwood xylan, respectively. XYN11A is a endo-β-1,4-xylanase since it release xylobiose and xylotriose as the main products by hydrolyzing xylans. The activity of XYN11A was enhanced 155 % by 1 mM Fe2+ ions, but was inhibited strongly by Fe3+. The reason of enhancing the xylanase activity of XYN11A with 1 mM Fe2+ treatment may be responsible for the change of microenvironment of tryptophan residues studied by synchronous fluorescence spectrophotometry. Inhibition of the xylanase activity by Fe3+ was first time demonstrated to associate tryptophan fluorescence quenching.

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Acknowledgments

This research was financially supported by the Agricultural Ministry of China (2011-G27), Special Fund for Agro-scientific Research in the Public Interest (201203001), National key technology R&D program (L020130249), and National Undergraduate Innovative Experiment Program (201310307020).

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Authors and Affiliations

  1. Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing, 210095, China

    Hanpeng Liao, Shaowei Sun, Pan Wang, Wenli Bi, Zhong Wei, Xinlan Mei, Dongyang Liu, Waseem Raza, Qirong Shen & Yangchun Xu

  2. Institute of Bast Fiber Crops and Center of Southern Economic Crops, Chinese Academy of Agricultural Sciences, Changsha, China

    Shiyong Tan

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  1. Hanpeng Liao
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Correspondence to Yangchun Xu.

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Liao, H., Sun, S., Wang, P. et al. A new acidophilic endo-β-1,4-xylanase from Penicillium oxalicum: cloning, purification, and insights into the influence of metal ions on xylanase activity. J Ind Microbiol Biotechnol 41, 1071–1083 (2014). https://doi.org/10.1007/s10295-014-1453-0

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