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Expression and characterisation of a pH and salt tolerant, thermostable and xylose tolerant recombinant GH43 β-xylosidase from Thermobifida halotolerans YIM 90462T for promoting hemicellulose degradation

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Abstract

A gene encoding a β-xylosidase (designated as Thxyl43A) was cloned from strain Thermobifida halotolerans YIM 90462T. The open reading frame of this gene encodes 550 amino acid residues. The gene was over-expressed in Escherichia coli and the recombinant protein was purified. The monomeric Thxyl43A protein presented a molecular mass of 61.5 kDa. When p-nitrophenyl-β-d-xylopyranoside was used as the substrate, recombinant Thxyl43A exhibited optimal activity at 55 °C and pH 4.0 to 7.0, being thermostable by maintaining 47% of its activity after 30 h incubation at 55 °C. The recombinant enzyme retained more than 80% residual activity after incubation at pH range of 4.0 to 12.0 for 24 h, respectively, which indicated notable thermostability and pH stability of Thxyl43A. Moreover, Thxyl43A displayed high catalytic activity (> 60%) in presence of 5–35% NaCl (w/v) or 1–20% ionic liquid (w/v) or 1–50 mM xylose. These properties suggest that Thxyl43A has potential for promoting hemicellulose degradation and other industrial applications.

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Acknowledgements

This research was supported by Infrastructure work project of China Ministry of Science and Technology (No. 2015FY110100), Guangzhou Municipal People’s Livelihood Science and Technology Plan (No. 201803030030), the Deanship of Scientific Research at Princess Nourah bint Abdulrahman University, through the Research Groups Program Grant No. (RGP-1438-0004) and China Postdoctoral Science Foundation (Grant No. 2017M622861). W-J Li is supported by project funded by Guangdong Province Higher Vocational Colleges & Schools Pearl River Scholar Funded Scheme (2014).

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Author notes

  1. Yi-Rui Yin and Wen-Dong Xian have been contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Biocontrol and Guangdong Provincial Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, People’s Republic of China

    Yi-Rui Yin, Wen-Dong Xian, Ming-Xian Han, En-Min Zhou, Lan Liu, Min Xiao & Wen-Jun Li

  2. Biology Department, Faculty of Science, Princess Nourah bint Abdulrahman University, Riyadh, 11564, Kingdom of Saudi Arabia

    Dalal Hussien M. Alkhalifah

  3. Bioproducts Research Chair, Zoology Department, College of Science, King Saud University, Riyadh, 11451, Kingdom of Saudi Arabia

    Wael N. Hozzein

  4. Botany and Microbiology Department, Faculty of Science, Beni-Suef University, Beni Suef, 62511, Egypt

    Wael N. Hozzein

  5. Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Ürümqi, 830011, China

    Wen-Jun Li

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  1. Yi-Rui Yin
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Contributions

Y.R.Y., M.X. and W.J.L. conceived the study. W.D.X. performed the culture of strains and collection of E. coli biomass. Y.R.Y. and M.X.H. performed the protein separation. E.M.Z., L.L and D.H.M.A. performed the measurement of enzyme activity. W.N.H. and M.X. performed the data analysis. Y.R.Y., W.D.X., M.X. and W.J.L. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Min Xiao or Wen-Jun Li.

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Yin, YR., Xian, WD., Han, MX. et al. Expression and characterisation of a pH and salt tolerant, thermostable and xylose tolerant recombinant GH43 β-xylosidase from Thermobifida halotolerans YIM 90462T for promoting hemicellulose degradation. Antonie van Leeuwenhoek 112, 339–350 (2019). https://doi.org/10.1007/s10482-018-1161-2

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