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Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli

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Abstract

In this study, phosphoenolpyruvate carboxylase (PEPC) derived from Oceanimonas smirnovii (OS) was expressed as a soluble protein in Escherichia coli BL21(DE3). We isolated OS-PEPC (a recombinant PEPC protein) by his-tag purification. The purified protein showed a single band upon analysis with SDS-PAGE, and it had an apparent molecular mass of 98 kDa. Pufied OS-PEPC showed a specific activity value of 21.8 ± 0.495 U/mg protein. Especially, OS-PEPC showed the enzymatic activity between 40 and 50 °C. It maintained enzymatic activity in basic pH conditions (pH value, 9–10). We also measured OS-PEPC PEP and HCO3 − saturation kinetics and confirmed the effect of divalent cation on OS-PEPC activity.

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Acknowledgments

This research was supported by the Basic Core Technology Development Program for the Oceans and the Polar Regions of the National Research Foundation (NRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2010-0020501).

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, 121-742, South Korea

    Soohyun Park, Wangjun Lee, Hyeonsoo Kim & Jinwon Lee

  2. Department of Biotechnology and Bioinformatics, Korea University, Sejong Campus, Sejong-ro 2511, Sejong, 339-700, South Korea

    Seung Pil Pack

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  1. Soohyun Park
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  2. Wangjun Lee
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  3. Hyeonsoo Kim
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  4. Seung Pil Pack
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  5. Jinwon Lee
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Correspondence to Seung Pil Pack or Jinwon Lee.

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Park, S., Lee, W., Kim, H. et al. Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli . Appl Biochem Biotechnol 177, 217–225 (2015). https://doi.org/10.1007/s12010-015-1739-3

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  • DOI: https://doi.org/10.1007/s12010-015-1739-3

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