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Immobilization and characterization of carbonic anhydrase purified from E. coli MO1 and its influence on CO2 sequestration


The present investigation entails the immobilisation and characterisation of Escherichia coli MO1-derived carbonic anhydrase (CA) and its influence on the transformation of CO2 to CaCO3. CA was purified from MO1 using a combination of Sephadex G-75 and DEAE cellulose column chromatography, resulting in 4.64-fold purification. The purified CA was immobilised in chitosan-alginate polyelectrolyte complex (C-A PEC) with an immobilisation potential of 94.5 %. Both the immobilised and free forms of the enzyme were most active and stable at pH 8.2 and at 37 °C. The K m and V max of the immobilised enzyme were found to be 19.12 mM and 416.66 μmol min−1 mg−1, respectively; whereas, the K m and V max of free enzyme were 18.26 mM and 434.78 μmol min−1 mg−1, respectively. The presence of metal ions such as Cu2+, Fe2+, and Mg2+ stimulated the enzyme activity. Immobilised CA showed higher storage stability and maintained its catalytic efficiency after repeated operational cycles. Furthermore, both forms of the enzyme were tested for targeted application of the carbonation reaction to convert CO2 to CaCO3. The amounts of CaCO3 precipitated over free and immobilised CA were 267 and 253 mg/mg of enzyme, respectively. The results of this study show that immobilised CA in chitosan-alginate beads can be useful for CO2 sequestration by the biomimetic route.

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First author gratefully acknowledges the Jawaharlal Nehru Memorial Fund (JNMF), New Delhi, for providing fellowship for doctoral study. Authors also thankful to Dr. S. Kumaran, Asst. Prof., Department of Biotechnology, Periyar Maniammai University, for his kind help in AFM study.

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Correspondence to Sib Sankar Giri.

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Oviya, M., Sukumaran, V. & Giri, S.S. Immobilization and characterization of carbonic anhydrase purified from E. coli MO1 and its influence on CO2 sequestration. World J Microbiol Biotechnol 29, 1813–1820 (2013).

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  • E. coli MO1
  • Carbonic anhydrase
  • Immobilisation
  • Chitosan-alginate
  • CO2 sequestration