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A Xylanase Gene Directly Cloned from the Genomic DNA of Alkaline Wastewater Sludge Showing Application Potential in the Paper Industry

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Abstract

A xylanase gene, aws-2x, was directly cloned from the genomic DNA of the alkaline wastewater sludge using degenerated PCR and modified TAIL-PCR. The deduced amino acid sequence of AWS-2x shared the highest identity (60%) with the xylanase from Chryseobacterium gleum belonging to the glycosyl hydrolase GH family 10. Recombinant AWS-2x was expressed in Escherichia coli BL21 (DE3) and purified to electrophoretic homogeneity. The enzyme showed maximal activity at pH 7.5 and 55 °C, maintained more than 50% of maximal activity when assayed at pH 9.0, and was stable over a wide pH range from 4.0 to 11.0. The specific activity of AWS-2x towards hardwood xylan (beechwood and birchwood xylan) was significantly higher than that to cereal xylan (oat spelt xylan and wheat arabinoxylan). These properties make AWS-2x a potential candidate for application in the pulp and paper industry.

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Acknowledgments

This research was supported by the Earmarked Fund for Modern Agro-industry Technology Research System (NYCYTX-42-G2-05) and the Key Program of Transgenic Plant Breeding (2009ZX08019-002) and the Agricultural Science and Technology Conversion Funds (2009GB23260444).

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Authors and Affiliations

  1. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Yanyu Zhao, Huiying Luo, Kun Meng, Pengjun Shi, Guozeng Wang, Tiezheng Yuan & Bin Yao

  2. Department of Microbial Engineering, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Yanyu Zhao & Peilong Yang

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  1. Yanyu Zhao
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  2. Huiying Luo
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  3. Kun Meng
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  4. Pengjun Shi
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  5. Guozeng Wang
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  6. Peilong Yang
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  7. Tiezheng Yuan
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  8. Bin Yao
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Correspondence to Peilong Yang or Bin Yao.

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Zhao, Y., Luo, H., Meng, K. et al. A Xylanase Gene Directly Cloned from the Genomic DNA of Alkaline Wastewater Sludge Showing Application Potential in the Paper Industry. Appl Biochem Biotechnol 165, 35–46 (2011). https://doi.org/10.1007/s12010-011-9231-1

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  • DOI: https://doi.org/10.1007/s12010-011-9231-1

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