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Molecular and biochemical characterization of a novel xylanase from the symbiotic Sphingobacterium sp. TN19

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Abstract

A xylanase-encoding gene, designated xynA19, was cloned from Sphingobacterium sp. TN19—a symbiotic bacterium isolated from the gut of Batocera horsfieldi larvae—and expressed in Escherichia coli BL21 (DE3). The full-length xynA19 (1,155 bp in length) encodes a 384-residue polypeptide (XynA19) containing a predicted signal peptide of 24 residues and a catalytic domain belonging to glycosyl hydrolase family 10 (GH 10). The deduced amino acid sequence of XynA19 is most similar (53.1% identity) to an endo-1,4-β-xylanase from Prevotella bryantii B14. Phylogenetic analysis of GH 10 Bacteroidia xylanases indicated that GH 10 xylanases from Sphingobacteria were separated into two clusters, and XynA19 is more closely related to the xylanases of Bacteroidia from gut or rumen than to those of Flavobacteria and Sphingobacteria from other sources. Recombinant XynA19 (r-XynA19) showed apparent optimal activity at pH 6.5 and 45°C. Compared with thermophilic and mesophilic counterparts, r-XynA19 was more active at low temperatures, retaining >65% of its maximum activity at 20–28°C and ~40% even at 10°C, and modeling indicated that XynA19 has fewer hydrogen bonds and salt bridges. These properties suggest that XynA19 has various potential applications, especially in aquaculture and the food industry.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program; No. 2007AA100601), the National Key Technology Research and Development Program of China (No. 2006BAD12B05-03), and the Chinese Program on Research for Public Good (Grant No. 2005DIB4J038). We are grateful to Professor Youqing Luo of Beijing Forestry University, China, for identification of Batocera horsfieldi.

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Correspondence to Bin Yao.

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Zhou, J., Huang, H., Meng, K. et al. Molecular and biochemical characterization of a novel xylanase from the symbiotic Sphingobacterium sp. TN19. Appl Microbiol Biotechnol 85, 323–333 (2009). https://doi.org/10.1007/s00253-009-2081-x

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Keywords

  • Batocera horsfieldi
  • Sphingobacterium sp. TN19
  • Xylanase
  • Gut