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Extremophiles

, Volume 13, Issue 5, pp 849–857 | Cite as

A thermophilic and acid stable family-10 xylanase from the acidophilic fungus Bispora sp. MEY-1

  • Huiying Luo
  • Jiang Li
  • Jun Yang
  • Hui Wang
  • Yuhui Yang
  • Huoqing Huang
  • Pengjun Shi
  • Tiezheng Yuan
  • Yunliu Fan
  • Bin YaoEmail author
Original Paper

Abstract

A complete gene, xyl10C, encoding a thermophilic endo-1,4-β-xylanase (XYL10C), was cloned from the acidophilic fungus Bispora sp. MEY-1 and expressed in Pichia pastoris. XYL10C shares highest nucleotide and amino acid sequence identities of 57.3 and 49.7%, respectively, with a putative xylanase from Aspergillus fumigatus Af293 of glycoside hydrolase family 10. A high expression level in P. pastoris (73,400 U ml−1) was achieved in a 3.7–l fermenter. The purified recombinant XYL10C was thermophilic, exhibiting maximum activity at 85°C, which is higher than that reported from any fungal xylanase. The enzyme was also highly thermostable, exhibiting ~100% of the initial activity after incubation at 80°C for 60 min and >87% of activity at 90°C for 10 min. The half lives of XYL10C at 80 and 85°C were approximately 45 and 3 h, respectively. It had two activity peaks at pH 3.0 and 4.5–5.0 (maximum), respectively, and was very acid stable, retaining more than 80% activity after incubation at pH 1.5−6.0 for 1 h. The enzyme was resistant to Co2+, Mn2+, Cr3+ and Ag+. The specific activity of XYL10C for oat spelt xylan was 18,831 U mg−1. It also had wide substrate specificity and produced simple products (65.1% xylose, 25.0% xylobiose and 9.9% xylan polymer) from oat spelt xylan.

Keywords

Thermophilic xylanase Acidophilic fungus Bispora sp. MEY-1 Pichia pastoris 

Notes

Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 program, Grant No. 2007AA100601), the Chinese Program on Research for Public Good (Grant No. 2005DIB4J038), and the 948 program of the Ministry of Agriculture (Grant No. 2007-Z3).

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Copyright information

© Springer 2009

Authors and Affiliations

  • Huiying Luo
    • 1
  • Jiang Li
    • 2
  • Jun Yang
    • 1
  • Hui Wang
    • 1
  • Yuhui Yang
    • 1
  • Huoqing Huang
    • 1
  • Pengjun Shi
    • 1
  • Tiezheng Yuan
    • 1
  • Yunliu Fan
    • 3
  • Bin Yao
    • 1
    Email author
  1. 1.Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research InstituteChinese Academy of Agricultural SciencesBeijingPeople’s Republic of China
  2. 2.Department of BiologyEast China Institute of TechnologyFuzhouPeople’s Republic of China
  3. 3.Biotechnology Research InstituteChinese Academy of Agricultural SciencesBeijingPeople’s Republic of China

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