Purification and characterization of phosphoenolpyruvate carboxylase from a cyanobacterium

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  • Phosphoenolpyruvate (PEP) carboxylase (EC was purified 100-fold from the cyanobacterium Coccochloris peniocystis with a yield of 10%. A single isozyme was found at all stages of purification, and activity of other beta-carboxylase enzymes was not detected. The apparent molecular weight of the native enzyme was 560,000. Optimal activity was observed at pH 8.0 and 40 degrees C, yielding a Vmax of 8.84 mumol/mg of protein per min. The enzyme was not protected from heat inactivation by aspartate, malate, or oxalacetate. Michaelis-Menten reaction kinetics were observed for various concentrations of PEP, Mg2+, and HCO3-, yielding Km values of 0.6, 0.27, and 0.8 mM, respectively. Enzyme activity was inhibited by aspartate and tricarboxylic acid cycle intermediates and noncompetitively inhibited by oxalacetate, while activation by any compound was not observed. However, the enzyme was sensitive to metabolic control at subsaturating substrate concentrations at neutral pH. These data indicate that cyanobacterial PEP carboxylase resembles the enzyme isolated from C3 plants (plants which initially incorporate CO2 into C3 sugars) and suggest that PEP carboxylase functions anapleurotically in cyanobacteria.

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    © 1986 George W. Owttrim et al. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
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    • Owttrim, George W., & Colman, Brian. (1986). Purification and characterization of phosphoenolpyruvate carboxylase from a cyanobacterium. Journal of bacteriology, 168(1), 207-212.
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