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Cell Surface Display and Characterization of Rhizopus oryzae Lipase in Pichia pastoris Using Sed1p as an Anchor Protein

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Abstract

It has been investigated to conduct the surface displaying of lipase from Rhizopus oryzae onto the cells of Pichia pastoris yeast using Sed1p as an anchor protein. A yeast cell surface display plasmid pPICZαA-rol-histag-sed1p was constructed by fusing rol and sed1p gene fragments into the plasmid pPICZαA, followed by introducing recombinant plasmid into P. pastoris cells. Surface display levels were monitored by Western Blot and immunofluorescence microscopy. The activity of displaying lipase obtained from recombinant mutS reached at 60 U/g-dry cell. In addition, the displaying lipase was stable in broad ranges of temperatures and pH, with the optimum temperature at 40 °C and pH 7.5. These results indicate that the P. pastoris displaying lipase may have potential in whole-cell biocatalyst.

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Acknowledgments

This work was financially supported by the Jiangsu Education (11KJA480001), the National Natural Science Foundation of China (No. 31170537), and the Doctorate Fellowship Foundation of Nanjing Forestry University.

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Authors and Affiliations

  1. College of Chemical Engineering, Nanjing Forestry University, Nanjing, China

    Wenqian Li, Hao Shi, Huaihai Ding, Liangliang Wang, Yu Zhang, Xun Li & Fei Wang

  2. Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, Nanjing, China

    Wenqian Li, Hao Shi, Huaihai Ding, Liangliang Wang, Yu Zhang, Xun Li & Fei Wang

  3. College of Life Science and Chemical Engineering, Huaiyin Institute of Technology, Huaian, China

    Wenqian Li

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  1. Wenqian Li
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  2. Hao Shi
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  4. Liangliang Wang
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  5. Yu Zhang
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  6. Xun Li
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  7. Fei Wang
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Correspondence to Fei Wang.

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Li, W., Shi, H., Ding, H. et al. Cell Surface Display and Characterization of Rhizopus oryzae Lipase in Pichia pastoris Using Sed1p as an Anchor Protein. Curr Microbiol 71, 150–155 (2015). https://doi.org/10.1007/s00284-015-0835-5

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  • DOI: https://doi.org/10.1007/s00284-015-0835-5

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