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Gene Cloning, Overexpression, and Characterization of a Xylanase from Penicillium sp. CGMCC 1669

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Abstract

A xylanase-encoding gene, xyn11F63, was isolated from Penicillium sp. F63 CGMCC1669 using degenerated polymerase chain reaction (PCR) and thermal asymmetric interlaced (TAIL)-PCR techniques. The full-length chromosomal gene consists of 724 bp, including a 73-bp intron, and encodes a 217 amino acid polypeptide. The deduced amino acid sequence of xyn11F63 shows the highest identity of 70% to the xylanase from Penicillium sp. strain 40, which belongs to glycosyl hydrolases family 11. The gene was overexpressed in Pichia pastoris, and its activity in the culture medium reached 516 U ml−1. After purification to electrophoretic homogeneity, the enzyme showed maximal activity at pH 4.5 and 40°C, was stable at acidic buffers of pH 4.5–9.0, and was resistant to proteases (proteinase K, trypsin, subtilisin A, and α-chymotrypsin). The specific activity, K m, and V max for oat spelt xylan substrate was 7,988 U mg−1, 22.2 mg ml−1, and 15,105.7 μmol min−1 mg−1, respectively. These properties make XYN11F63 a potential economical candidate for use in feed and food industrial applications.

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Acknowledgements

This work was supported by the Chinese National High Technology Research and Development Program (863 Program, Grant No. 2007AA100601) and the Chinese Agricultural Microorganism Collection and Share Program (No. 2005DKA21201).

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Authors and Affiliations

  1. Key Laboratory of 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

    Wanli Liu, Pengjun Shi, Peilong Yang, Guozeng Wang, Yaru Wang, Huiying Luo & Bin Yao

  2. Institute of Biochemistry and Molecular Biology, College of Life Science, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Wanli Liu & Qiang Chen

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  1. Wanli Liu
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  2. Pengjun Shi
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  3. Qiang Chen
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  5. Guozeng Wang
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  6. Yaru Wang
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  7. Huiying Luo
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  8. Bin Yao
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Correspondence to Bin Yao.

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Liu, W., Shi, P., Chen, Q. et al. Gene Cloning, Overexpression, and Characterization of a Xylanase from Penicillium sp. CGMCC 1669. Appl Biochem Biotechnol 162, 1–12 (2010). https://doi.org/10.1007/s12010-009-8719-4

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  • DOI: https://doi.org/10.1007/s12010-009-8719-4

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