title: Engineering de novo disulfide bond in bacterial α-type carbonic anhydrase for thermostable carbon sequestration.
abstract: Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial α-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 °C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions.
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors.forename: Byung Hoon
authors.initials: BH
authors.lastname: Jo
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors.forename: Tae Yoon
authors.initials: TY
authors.lastname: Park
authors.affiliation: Bio-Max Institute, Seoul National University, Seoul 151-742, Korea.
authors.forename: Hyun June
authors.initials: HJ
authors.lastname: Park
authors.affiliation: The Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea.
authors.forename: Young Joo
authors.initials: YJ
authors.lastname: Yeon
authors.affiliation: Bio-Max Institute, Seoul National University, Seoul 151-742, Korea.|School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea.
authors.forename: Young Je
authors.initials: YJ
authors.lastname: Yoo
authors.affiliation: Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Cha
mesh_terms: D057965:Carbon Sequestration / Q000502:physiology*; D002256:Carbonic Anhydrases / Q000378:metabolism*; D020134:Catalytic Domain / Q000502:physiology; D004220:Disulfides / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D013696:Temperature; D013816:Thermodynamics
chemical_list: D004220:Disulfides; D002256:Carbonic Anhydrases