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Applied Microbiology and Biotechnology

, Volume 103, Issue 21–22, pp 8899–8909 | Cite as

Cloning and heterologous expression of a novel halo/alkali-stable multi-domain xylanase (XylM18) from a marine bacterium Marinimicrobium sp. strain LS-A18

  • Hao Yu
  • Shuxue Zhao
  • Yaqin Fan
  • Chunhui Hu
  • Weidong LuEmail author
  • Lizhong GuoEmail author
Biotechnologically relevant enzymes and proteins
  • 75 Downloads

Abstract

Halophilic bacteria are good bioresources for halotolerant alkaline enzymes. A multi-domain high-molecular-weight endo-β-1,4-xylanase gene, xylM18, was cloned from a halophilic marine bacterium Marinimicrobium sp. LS-A18. XylM18 is different from any of the functionally reported xylanases. It has a glycosyl hydrolase (GH) 43 domain, a GH10 domain, and two serine-rich linkers, representing a novel family. The gene, encoding 1022 residues, was cloned and heterologously expressed in Escherichia coli BL21(DE3) cells. Purified XylM18 was proved to be a xylanase. It showed diminished activity without salt and showed activity with a broad NaCl range from 0.2 to 25% (w/v). NaCl can increase the optimal temperature from 30 °C (0% NaCl) to 50 °C (10% NaCl). The purified XylM18 was active between pH 6.0 and 10.0 and was optimally active at pH 7.0. The xylanase activities were basically unchanged at a NaCl concentration range from 10 to 20% or pH from 7 to 10 after 24 h incubation. The apparent Km and Vmax values of XylM18 for xylan were 2.76 mg/mL and 60.0 U/mg, respectively. The GH10 domain of this enzyme, XylM18-GH10, was expressed and characterized. XylM18-GH10 also showed xylanase activity and maintained halo-stable property. The apparent Km and Vmax values of XylM18-GH10 for xylan were 1.60 mg/mL and 130.1 U/mg, respectively. Other domains of XylM18 showed no xylanase activity. In summary, XylM18 is a halo-tolerant and alkali-stable endoxylanase which is a suitable candidate for xylan biodegradation in high-salt and alkali conditions. To our knowledge, this is the first report of a multidomain high-molecular-weight xylanase.

Keywords

Endo-β-1,4-xylanase Glycosyl hydrolase family 10 Halophilic bacterium Marinimicrobium 

Notes

Acknowledgments

This study was funded by the Shandong Provincial Natural Science Foundation (ZR2016CQ06), Key Project of Edible Fungus Genetic Breeding System of Modern Agriculture of Shandong Province, and the Qingdao Agricultural University Scientific Research Foundation (6631115052). We are very grateful to Dr Joel Rankin (Michigan State University) for valuable comments on the manuscript.

Author contributions

H.Y., S.Z., W.L., and L.G. conceived and designed the project. H. Y., S.Z., and Y.F. performed the experiments. H.Y., W.L., and L.G. contributed reagents and materials. H.Y., C.H., Y.F., and S.Z. analyzed data. H.Y., C.H., and L.G. wrote the manuscript. All of the authors have read and approved the manuscript.

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.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Shandong Provincial Key Laboratory of Applied Mycology, College of Life SciencesQingdao Agricultural UniversityQingdaoPeople’s Republic of China

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