Applied Microbiology and Biotechnology

, Volume 102, Issue 8, pp 3675–3685 | Cite as

Engineering improved thermostability of the GH11 xylanase from Neocallimastix patriciarum via computational library design

  • Yifan Bu
  • Yinglu Cui
  • Ying Peng
  • Meirong Hu
  • Yu’e Tian
  • Yong Tao
  • Bian Wu
Biotechnologically relevant enzymes and proteins
  • 215 Downloads

Abstract

Xylanases, which cleave the β-1,4-glycosidic bond between xylose residues to release xylooligosaccharides (XOS), are widely used as food additives, animal feeds, and pulp bleaching agents. However, the thermally unstable nature of xylanases would hamper their industrial application. In this study, we used in silico design in a glycoside hydrolase family (GH) 11 xylanase to stabilize the enzyme. A combination of the best mutations increased the apparent melting temperature by 14 °C and significantly enhanced thermostability and thermoactivation. The variant also showed an upward-shifted optimal temperature for catalysis without compromising its activity at low temperatures. Moreover, a 10-fold higher XOS production yield was obtained at 70 °C, which compensated the low yield obtained with the wild-type enzyme. Collectively, the variant constructed by the computational strategy can be used as an efficient biocatalyst for XOS production at industrially viable conditions.

Keywords

Xylanase XOS Xylan FRESCO Thermostability 

Notes

Acknowledgements

This work is supported by the National Natural Science Foundation of China (Grant Nos. 21603013, 31601412), the 100 Talent Program grant, and Biological Resources Service Network Initiative (ZSYS-012) and grant (SKT1604) from the Chinese Academy of Sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

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

Supplementary material

253_2018_8872_MOESM1_ESM.pdf (563 kb)
ESM 1 (PDF 562 kb)

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Copyright information

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Authors and Affiliations

  1. 1.CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of MicrobiologyChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.State Key Laboratory of Transducer TechnologyChinese Academy of SciencesBeijingPeople’s Republic of China

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