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.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Abu-Khader M (2006) Recent progress in CO2 capture/sequestration: a review. Energy Sour Part A Recover Util Environ Effects 28:1261–1279
Ansari SA, Husain Q (2011) Immobilization of Kluyveromyces lactis β galactosidase on concanavalin A layered aluminium oxide nanoparticles—its future aspects in biosensor applications. J Mol Catal B Enzym 70:119–126
Bertini I, Luchinat C, Monnanni R, Roelens S, Moratal JM (1987) Interaction of carbon dioxide and copper (II) carbonic anhydrase. J Am Chem Soc 109:7855–7856
Bond GM, Stringer J, Brandvold DK, Simsek FA, Medina MG, Egeland G (2001) Development of integrated system for biomimetic CO2 sequestration using the enzyme carbonic anhydrase. Energy Fuels 15:309–316
Camacho-Rubio F, Jurado-Alameda E, Gonzalez-Tello P, Luzon GGA (1996) Comparative study of the activity of free and immobilized enzymes and its application to glucose isomerase. Chem Eng Sci 51:4159–4165
Chart H, Smith HR, La Ragione RM, Woodward MJ (2000) An investigation into the pathogenic properties of Escherichia coli strains BLR, BLR21, DH5α and EQ1. J Appl Microbiol 89:1048–1058
Favre N, Christ ML, Pierre AC (2009) Biocatalytic capture of CO2 with carbonic anhydrase and its transformation to solid carbonate. J Mol Catal B Enzym 60:163–170
Hanley SJ, Giasson J, Revol JF, Gray DG (1992) Atomic force microscopy of cellulose microfibrils: comparison with transmission electron microscopy. Polymer 33:4639–4642
Heck RW, Tanhouser SM, Manda R, Tu C, Laipis PJ, Silverman DN (1994) Catalytic properties of mouse carbonic anhydrase V. J Biol Chem 269:24742–24746
Karlsson J, Clarke AK, Chen ZY, Hugghins SY, Park Y, Husic HD, Moroney JV, Samuelsson G (1998) A novel α-type carbonic anhydrase associated with the thylakoid membrane in Clamydomonas reinhardtii is required for growth at ambient CO2. EMBO J 17(5):1208–1216
Keeling CD, Whorf TP, Wahlen M, Van der Pllcht J (1995) Interannual extremes in the rate of rise of atmospheric carbon dioxide since 1980. Nature 375:666–670
Kotwica J, Ciuk MA, Joachimiak E, Rowinski S, Cymborowiski PB (2006) Carbonic anhydrase activity in the vas deferens of the of the cotton leaf worm–Spodopetra littoralis (Lepidoptera: Noctuidae) controlled by circadian clock. J Physiol Pharmacol 57(8):107–123
Krajewska B (2004) Application of chitin- and chitosan-based materials for enzyme immobilizations: a review. Enzym Microb Technol 35:126–139
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lowry OH, Rosebbrough NJ, Farr AL, Randall RL (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–273
Oviya M, Giri SS, Sukumaran V, Natarajan P (2012) Immobilization of carbonic anhydrase enzyme purified from Bacillus subtilis VSG-4 and its application as CO2 sequester. Prep Biochem Biotech 42:462–475
Prabhu C, Wanjari S, Gawande S, Das S, Labhsetwar N, Kotwal S, Puri AK, Satyanarayana T, Rayalu S (2009) Immobilization of carbonic anhydrase enriched microorganism on biopolymer based materials. J Mol Catal B Enzym 60:13–21
Prabhu C, Valechha A, Wanjari S, Labhsetwar N, Kotwal S, Satyanarayanan T, Rayalu S (2011) Carbon composite beads for immobilization of carbonic anhydrase. J Mol Catal B Enzym 71:71–78
Ramanan R, Kannan K, Sivanesan SD, Mudliar S, Kaur S, Tripathi AK, Chakrabarti T (2009a) Bio-sequestration of carbon dioxide using carbonic anhydrase enzyme purified from Citrobacter freundii. World J Microbiol Biotechnol 25:981–987
Ramanan R, Kannan K, Vinayagamoorthy N, Ramkumar KM, Sivanesan SD, Chakrabarti T (2009b) Purification and characterization of a novel plant-type carbonic anhydrase from Bacillus subtilis. Biotechnol Bioprocess Eng 14:32–37
Reifert H (2007) Anthropogenic change-CO2 rising faster. Proc Natl Acad Sci USA 104:10288–10293
Sahin F, Demirel G, Tumturk H (2005) A novel matrix for the immobilization of acetylcholinesterase. Int J Biol Macromol 37:148–153
Sharma A, Bhattacharya A (2010) Enhanced biomimetic sequestration of CO2 into CaCO3 using purified carbonic anhydrase from indigenous bacterial strains. J Mol Catal B Enzym 67:122–128
Sharma A, Bhattacharya A, Shrivastava A (2011) Biomimetic CO2 sequestration using purified carbonic anhydrase from indigenous bacterial strains immobilized on biopolymeric materials. Enzym Microbiol Technol 48:416–426
Smith KS, Ferry JG (2000) Prokaryotic carbonic anhydrases. FEMS Microbiol Rev 24:335–366
Tripp BC, Bell CB, Cruz F, Krebs C, Ferry JG (2004) A role for iron in an ancient carbonic anhydrase. J Biol Chem 279:6683–6687
Vu TKH, Le VVM (2008) Biochemical studies on the immobilization of the enzyme invertase (EC 3. 2. 1. 26) in alginate gel and its kinetics. ASEAN Food J 15(1):73–78
Wanjari S, Prabhu C, Yadav R, Satyanarayana T, Labhsetwar N (2011) Immobilization of carbonic anhydrase on chitosan beads for enhanced carbonation reaction. Process Biochem 46:1010–1018
Wilbur KM, Anderson NG (1948) Electrometric and calorimetric determination of carbonic anhydrase. J Biol Chem 176:147–154
Yadav RR, Mudliar SN, Shekh AY, Fulke AB, Devi SS, Krishnamurthi K, Juwarkar A, Chakrabarti T (2012) Immobilization of carbonic anhydrase in alginate and its influence on transformation of CO2 to calcite. Process Biochem 47:585–590
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.
About this article
Cite this article
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). https://doi.org/10.1007/s11274-013-1343-z
- E. coli MO1
- Carbonic anhydrase
- CO2 sequestration