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Applied Microbiology and Biotechnology

, Volume 98, Issue 5, pp 2155–2163 | Cite as

A novel thermophilic endo-β-1,4-mannanase from Aspergillus nidulans XZ3: functional roles of carbohydrate-binding module and Thr/Ser-rich linker region

  • Haiqiang Lu
  • Huiying Luo
  • Pengjun Shi
  • Huoqing Huang
  • Kun Meng
  • Peilong Yang
  • Bin YaoEmail author
Biotechnologically relevant enzymes and proteins

Abstract

The gene man5XZ3 from Aspergillus nidulans XZ3 encodes a multimodular β-mannanase of glycoside hydrolase family 5 that consists of a family 1 carbohydrate-binding module (CBM1), a Thr/Ser-rich linker region, and a catalytic domain. Recombinant Man5XZ3 and its two truncated derivatives, Man5ΔCBM (removing the CBM1) and Man5ΔCL (removing both the CBM1 and linker region), were produced in Pichia pastoris and showed significant variance in the secondary structure. The three enzymes had similar biochemical properties, such as optimal pH and temperature (pH 5.0 and 80 °C) and excellent pH stability at pH 4.0–10.0. Removal of the CBM1 alone could improve the thermostability of Man5XZ3, but further removal of the linker region resulted in worse thermostability. Man5XZ3 retained greater enzyme activity in the presence of an organic solvent (acetone), two detergents (SDS and Triton X-100), and a chaotropic agent (urea) compared with Man5ΔCBM and Man5ΔCL. This study provides an excellent β-mannanase candidate favorable for various industries and primarily demonstrates the relationship between enzyme structure and function.

Keywords

β-Mannanase Thermophilic CBM Linker region Thermostability 

Notes

Acknowledgments

This research was supported by the National Science Foundation for Distinguished Young Scholars of China (31225026), the National High Technology Research and Development Program of China (863 Program; No. 2012AA022208), and the China Modern Agriculture Research System (CARS-42).

Supplementary material

253_2013_5112_MOESM1_ESM.pdf (151 kb)
ESM 1 (PDF 151 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Haiqiang Lu
    • 1
  • Huiying Luo
    • 1
  • Pengjun Shi
    • 1
  • Huoqing Huang
    • 1
  • Kun Meng
    • 1
  • Peilong Yang
    • 1
  • 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

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