title: Expression, reconstruction and characterization of codon-optimized carbonic anhydrase from Hahella chejuensis for CO2 sequestration application.
abstract: The high production of functional carbonic anhydrase (CA) is required for practical CO2 sequestration application mediated by CA. Here, the synthetic gene based on Escherichia coli codon usage of new α-type CA (HC-aCA) of Hahella chejuensis, a Korea marine microorganism, was highly expressed in E. coli. We obtained a high yield of functional HC-aCA by denaturing/refolding process and incorporating zinc ion into its active site. The refolded HC-aCA displayed a half-deactivation temperature of 60 °C with maximal activity at 50 °C, and had high pH stability in alkali condition with maximal activity at pH 10.0. The esterase activity of HC-aCA almost doubled at high salt concentration ranging from 0.67 to 2.0 M NaCl. HC-aCA catalyzed the conversion of CO2 to CaCO3 as calcites form in the presence of Ca(2+). The refolded HC-aCA could be a promising candidate for the development of efficient CA-based CO2 sequestration processes.
authors: Department of Biotechnology and Bioinformatics, Korea University, Jochiwon, Chungnam, 339-800, Korea.
authors: Mi-Ran
authors: MR
authors: Ki
authors: Kiha
authors: K
authors: Min
authors: Bashistha Kumar
authors: BK
authors: Kanth
authors: Jinwon
authors: J
authors: Lee
authors: Seung Pil
authors: SP
authors: Pack
mesh_terms: D002245:Carbon Dioxide / Q000737:chemistry*; D002256:Carbonic Anhydrases / Q000096:biosynthesis / Q000737:chemistry*; D002384:Catalysis; D020134:Catalytic Domain; D003062:Codon; D004926:Escherichia coli / Q000378:metabolism; D004950:Esterases / Q000737:chemistry; D020563:Gammaproteobacteria / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D015032:Zinc / Q000737:chemistry
chemical_list: D003062:Codon; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D002245:Carbon Dioxide; D004950:Esterases; D002256:Carbonic Anhydrases; D015032:Zinc