title: Cloning and expression of gamma carbonic anhydrase from Serratia sp. ISTD04 for sequestration of carbon dioxide and formation of calcite.
abstract: Bacterial strains isolated from marble mines rock and enriched in the chemostat culture with different concentrations of sodium bicarbonate. The enriched consortium had six bacterial isolates. One of bacterium isolate showed carbonic anhydrase (CA) activity by catalyzing the reversible hydration reaction of carbon dioxide to bicarbonate. The bacterium was identified as Serratia sp. by 16S rRNA sequence analysis. The carbonic anhydrase gene from Serratia sp. was found to be homologous with gamma carbonic anhydrase. The carbonic anhydrase gene was cloned in PET21b(+) and expressed it in recombinant Escherichia coli BL21 (DE3) with His-tag at the C-terminus. The recombinant protein was purified efficiently by using one-step nickel affinity chromatography. Expected size of carbonic anhydrase was approximately 29 kDa in SDS-PAGE gel. Recombinant carbonic anhydrase enzyme was used for biomineralization-based conversion of atmospheric CO2 into valuable calcite minerals. The calcification was confirmed by using XRD, FTIR, EDX and SEM analysis.
authors: School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110067, India; Amity School of Earth and Environmental Science, Amity University Gurgaon, India.
authors: Shaili
authors: S
authors: Srivastava
authors: School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110067, India.
authors: Randhir Kumar
authors: RK
authors: Bharti
authors: National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi, India.
authors: Praveen Kumar
authors: PK
authors: Verma
authors: School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110067, India. Electronic address: isthakur@hotmail.com.
authors: Indu Shekhar
authors: IS
authors: Thakur
mesh_terms: D002119:Calcium Carbonate / Q000737:chemistry*; D002245:Carbon Dioxide / Q000737:chemistry*; D057965:Carbon Sequestration; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D057232:Global Warming; D008906:Mining; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012705:Serratia / Q000201:enzymology*; D017693:Sodium Bicarbonate / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D014961:X-Ray Diffraction
chemical_list: D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D002245:Carbon Dioxide; D017693:Sodium Bicarbonate; D002256:Carbonic Anhydrases; D002119:Calcium Carbonate
keywords: CO(2) sequestration; Carbonic anhydrase; FTIR; Serratia sp.; XRD