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High expression of recombinant Streptomyces sp. S38 xylanase in Pichia pastoris by codon optimization and analysis of its biochemical properties

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Abstract

In recent years, the biotechnological use of xylanases has grown remarkably. To efficiently produce xylanase for food processing and other industry, a codon-optimized recombinant xylanase gene from Streptomyces sp. S38 was synthesized and extracellularly expressed in Pichia pastoris under the control of AOX1 promoter. SDS-PAGE and activity assay demonstrated that the molecular mass of the recombinant xylanase was estimated to be 25 kDa, the optimum pH and optimum temperature were 5.5 and 50°C, respectively. In shake flask culture, the specific activity of the xylanase activity was 5098.28 U/mg. The K m and V max values of recombinant xylanase were 11.0 mg/ml and 10000 μmol min−1 mg−1, respectively. In the presence of metal ions such as Ca2+, Cu2+, Cr3+ and K+, the activity of the enzyme increased. However, strong inhibition of the enzyme activity was observed in the presence of Hg2+. This is the first report on the expression properties of a recombinant xylanase gene from the Streptomyces sp. S38 using Pichia pastoris. The attractive biochemical properties of the recombinant xylanase suggest that it may be a useful candidate for variety of commercial applications.

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Acknowledgments

The research was supported by the Youth Fund of Shanghai Academy of Agricultural Sciences (2008-6,2009-19); Development Foundation of Shanghai Academy of Agricultural Sciences (2009-09); The Shanghai Basic Reseach Project (08JC1418000); The Key Project Fund of the Shanghai Municipal Committee of Agriculture (No. 2008-7-5).

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  1. Agro-Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd, Shanghai, 201106, China

    Xiao-Yan Fu, Wei Zhao, Ai-Sheng Xiong, Yong-Sheng Tian & Ri-He Peng

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  1. Xiao-Yan Fu
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  2. Wei Zhao
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  3. Ai-Sheng Xiong
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  4. Yong-Sheng Tian
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Correspondence to Ri-He Peng.

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Xiao-Yan Fu and Wei Zhao contributed equally to the article.

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Fu, XY., Zhao, W., Xiong, AS. et al. High expression of recombinant Streptomyces sp. S38 xylanase in Pichia pastoris by codon optimization and analysis of its biochemical properties. Mol Biol Rep 38, 4991–4997 (2011). https://doi.org/10.1007/s11033-010-0644-7

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  • DOI: https://doi.org/10.1007/s11033-010-0644-7

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