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The N-domain of Escherichia coli phosphoglycerate kinase is a novel fusion partner to express aggregation-prone heterologous proteins

Authors

  • Jong-Am Song,

    1. Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102
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  • Dae-Sung Lee,

    1. Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102
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  • Jin-Seung Park,

    1. Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102
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  • Kyung-Yeon Han,

    1. Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102
    Current affiliation:
    1. Emerging Technology Research Center, Corporate Technology Operations SAIT, Samsung Electronics Co., Ltd., Giheung, Republic of Korea.
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  • Jeewon Lee

    Corresponding author
    1. Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102
    • Department of Chemical and Biological Engineering, Korea University, 136-713 Anam-Dong 5-1, Seoul, Republic Korea, telephone: +82-2-3290-3304; fax: +82-2-926-6102.
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  • Jong-Am Song and Dae-Sung Lee contributed equally to this work.

Abstract

As a fusion partner to express aggregation-prone heterologous proteins, we investigated the efficacy of Escherichia coli phosphoglycerate kinase (ePGK) that consists of two functional domains (N- and C-domain) and reportedly has a high structural stability. When the full-length ePGK (F-ePGK) was used as a fusion partner, the solubility of the heterologous proteins increased, but some of them still had a large fraction of insoluble aggregates. Surprisingly, the fusion expression using the N-domain of ePGK (N-ePGK) made the insoluble fraction significantly reduce to less than 10% for all the heterologous fusion proteins tested. Also, we evaluated the efficacy of N-ePGK in making the target proteins be expressed with their own native function or structure. It was found that of human ferritin light chain, bacterial arginine deiminase, human granulocyte colony stimulating factor were synthesized evidently with the self-assembly function, L-arginine-degrading activity, and the correct secondary structure, respectively, through the fusion expression using N-ePGK. These results indicate that N-ePGK is a highly potent fusion partner that can be widely used for the synthesis of a variety of heterologous proteins in E. coli. Biotechnol. Bioeng. 2012; 109:325–335. © 2011 Wiley Periodicals, Inc.

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