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Gene cloning and expression of a new acidic family 7 endo-β-1,3-1,4-glucanase from the acidophilic fungus Bispora sp. MEY-1

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Abstract

Most reported microbial β-1,3-1,4-glucanases belong to the glycoside hydrolase family 16. Here, we report a new acidic family 7 endo-β-1,3-1,4-glucanase (Bgl7A) from the acidophilic fungus Bispora sp. MEY-1. The cDNA of Bgl7A was isolated and over-expressed in Pichia pastoris, with a yield of about 1,000 U ml–1 in a 3.7-l fermentor. The purified recombinant Bgl7A had three activity peaks at pH 1.5, 3.5, and 5.0 (maximum), respectively, and a temperature optimum at 60°C. The enzyme was stable at pH 1.0–8.0 and highly resistant to both pepsin and trypsin. Belonging to the group of non-specific endoglucanase, Bgl7A can hydrolyze not only β-glucan and cellulose but also laminarin and oat spelt xylan. The specific activity of Bgl7A against barley β-glucan and lichenan (4,040 and 2,740 U mg–1) was higher than toward carboxymethyl cellulose sodium (395 U mg–1), which was different from other family 7 endo-β-glucanases.

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Acknowledgments

This research was supported by the National High Technology Research and Development Program of China (863 program, grant no. 2007AA100601), Chinese Program on Research for Public Good (grant no. 2005DIB4J038), and 948 program of the Ministry of Agriculture (grant no. 2007-Z3).

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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

    Huiying Luo, Jun Yang, Peilong Yang, Huoqing Huang, Pengjun Shi, Yingguo Bai, Yaru Wang & Bin Yao

  2. Department of Biology, East China Institute of Technology, Fuzhou, Jiangxi, 344000, People’s Republic of China

    Jiang Li

  3. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Yunliu Fan

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  1. Huiying Luo
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  2. Jun Yang
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  3. Peilong Yang
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  4. Jiang Li
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  5. Huoqing Huang
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  6. Pengjun Shi
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  7. Yingguo Bai
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  8. Yaru Wang
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  9. Yunliu Fan
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  10. Bin Yao
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Correspondence to Bin Yao.

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Huiying Luo and Jun Yang contributed equally to this work.

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Luo, H., Yang, J., Yang, P. et al. Gene cloning and expression of a new acidic family 7 endo-β-1,3-1,4-glucanase from the acidophilic fungus Bispora sp. MEY-1. Appl Microbiol Biotechnol 85, 1015–1023 (2010). https://doi.org/10.1007/s00253-009-2119-0

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