Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli
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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.
KeywordsOceanimonas smirnovii Phosphoenolpyruvate carboxylase Characterization Bicarbonate
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).
- 8.Wang, D., Li, Q., Li, W. L., Xing, J. M., & Su, Z. Q. (2009). Improvement of succinate production by overexpression of a cyanobacterial carbonic anhydrase in Escherichia coli Enzym. Microbial. Technol., 45, 491–497.Google Scholar
- 9.Park, S. H., Hong, S. H., Pack, S. P., & Lee, J. W. (2014). High activity and stability of codon-optimized phosphoenolpyruvate carboxylase from Photobacterium profundum SS9 at low temperatures and its application for in vitro production of oxaloacetate. Bioprocess and Biosystems Engineering, 37, 331–335.CrossRefGoogle Scholar
- 16.Kai, Y., Matsumura, H., Inoue, T., Terada, K., Nagara, Y., Yoshinaga, T., Kihara, A., Tsumura, K., & Izui, K. (1999). Three-dimensional structure of phosphoenolpyruvate carboxylase: a proposed mechanism for allosteric inhibition. Proceedings of the National Academy of Sciences, 96, 823–828.CrossRefGoogle Scholar