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Cloning, Expression, and Characterization of an GHF 11 Xylanase from Aspergillus niger XZ-3S

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Abstract

A xylanase gene (xynZF-2) from the Aspergillus niger XZ-3S was cloned and expressed in Escherichia coli. The coding region of the gene was separated by only one intron with the 68 bp in length. It encoded 225 amino acid residues of a protein with a calculated molecular weight of 24.04 kDa plus a signal peptide of 18 amino acids. The amino acid sequence of the xynZF-2 gene had a high similarity with those of family 11 of glycosyl hydrolases reported from other microorganisms. The mature peptide encoding cDNA was subcloned into pET-28a(+) expression vector. The resultant recombinant plasmid pET-28a-xynZF-2 was transformed into E. coli BL21(DE3), and finally the recombinant strain BL21/xynZF-2 was obtained. A maximum activity of 42.33 U/mg was gained from cellular of E. coli BL21/xynZF-2 induced by IPTG. The optimum temperature and pH for recombinant enzyme which has a good stability in alkaline conditions were 40 °C and 5.0, respectively. Fe3+ had an active effect on the enzyme obviously.

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Acknowledgments

This study was financially supported by the Foundation of He’nan Educational Committee (No. 2011A180026), the Foundation of He’nan Science and Technology Agency of China (No. 112102210299) and the Foundation of He’nan Province of China for Key Young Teacher in University (No. 2011GGJS-125).

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Authors and Affiliations

  1. Department of Life Science and Technology, Xinxiang Medical University, 601 Jinsui Road, Xinxiang, 453003, People’s Republic of China

    Guanhua Fu, Dandan Wang & Chenyan Zhou

  2. The First Affiliated Hospital, Xinxiang Medical University, Jiankang Road, Weihui, 453100, People’s Republic of China

    Yongtao Wang

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  1. Guanhua Fu
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  2. Yongtao Wang
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  3. Dandan Wang
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  4. Chenyan Zhou
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Correspondence to Chenyan Zhou.

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Fu, G., Wang, Y., Wang, D. et al. Cloning, Expression, and Characterization of an GHF 11 Xylanase from Aspergillus niger XZ-3S. Indian J Microbiol 52, 682–688 (2012). https://doi.org/10.1007/s12088-012-0314-7

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  • DOI: https://doi.org/10.1007/s12088-012-0314-7

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