TY - JOUR
T1 - Stability-Controllable Self-Immobilization of Carbonic Anhydrase Fused with a Silica-Binding Tag onto Diatom Biosilica for Enzymatic CO2Capture and Utilization
AU - Kim, Suhyeok
AU - Joo, Kye Il
AU - Jo, Byung Hoon
AU - Cha, Hyung Joon
N1 - Funding Information:
This work was supported by the Basic Core Technology Development Program for the Oceans and the Polar Regions (NRF-2015M1A5A1037055) of the National Research Foundation funded by the Ministry of Science and ICT, Korea (to H.J.C.), the Korea Electric Power Research Institute grant (to H.J.C.), the Korea Institute of Energy Technology Evaluation and Planning grant (20182010600430) funded by the Ministry of Trade, Industry & Energy, Korea (to B.H.J.), and the National Research Foundation grant (NRF-2019R1F1A1063181) funded by the Ministry of Science and ICT, Korea (to B.H.J.).
Funding Information:
This work was supported by the Basic Core Technology Development Program for the Oceans and the Polar Regions (NRF-2015M1A5A1037055) of the National Research Foundation funded by the Ministry of Science and ICT, Korea (to H.J.C.), the Korea Electric Power Research Institute grant (to H.J.C.), the Korea Institute of Energy Technology Evaluation and Planning grant (20182010600430) funded by the Ministry of Trade Industry & Energy, Korea (to B.H.J.), and the National Research Foundation grant (NRF-2019R1F1A1063181) funded by the Ministry of Science and ICT, Korea (to B.H.J.).
Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/6/17
Y1 - 2020/6/17
N2 - Exploiting carbonic anhydrase (CA), an enzyme that catalyzes the hydration of CO2, is a powerful route for eco-friendly and cost-effective carbon capture and utilization. For successful industrial applications, the stability and reusability of CA should be improved, which necessitates enzyme immobilization. Herein, the ribosomal protein L2 (Si-tag) from Escherichia coli was utilized for the immobilization of CA onto diatom biosilica, a promising renewable support material. The Si-tag was redesigned (L2NC) and genetically fused to CA from the marine bacterium Hydrogenovibrio marinus (hmCA). One-step self-immobilization of hmCA-L2NC onto diatom biosilica by simple mixing was successfully achieved via Si-tag-mediated strong binding, showing multilayer adsorption with a maximal loading of 1.4 wt %. The immobilized enzyme showed high reusability and no enzyme leakage even under high temperature conditions. The activity of hmCA-L2NC was inversely proportional to the enzyme loading, while the stability was directly proportional to the enzyme loading. This discovered activity-stability trade-off phenomenon could be attributed to macromolecular crowding on the highly dense surface of the enzyme-immobilized biosilica. Collectively, our system not only facilitates the stability-controllable self-immobilization of enzyme via Si-tag on a diatom biosilica support for the robust, facile, and green construction of stable biocatalysts, but is also a unique model for studying the macromolecular crowding effect on surface-immobilized enzymes.
AB - Exploiting carbonic anhydrase (CA), an enzyme that catalyzes the hydration of CO2, is a powerful route for eco-friendly and cost-effective carbon capture and utilization. For successful industrial applications, the stability and reusability of CA should be improved, which necessitates enzyme immobilization. Herein, the ribosomal protein L2 (Si-tag) from Escherichia coli was utilized for the immobilization of CA onto diatom biosilica, a promising renewable support material. The Si-tag was redesigned (L2NC) and genetically fused to CA from the marine bacterium Hydrogenovibrio marinus (hmCA). One-step self-immobilization of hmCA-L2NC onto diatom biosilica by simple mixing was successfully achieved via Si-tag-mediated strong binding, showing multilayer adsorption with a maximal loading of 1.4 wt %. The immobilized enzyme showed high reusability and no enzyme leakage even under high temperature conditions. The activity of hmCA-L2NC was inversely proportional to the enzyme loading, while the stability was directly proportional to the enzyme loading. This discovered activity-stability trade-off phenomenon could be attributed to macromolecular crowding on the highly dense surface of the enzyme-immobilized biosilica. Collectively, our system not only facilitates the stability-controllable self-immobilization of enzyme via Si-tag on a diatom biosilica support for the robust, facile, and green construction of stable biocatalysts, but is also a unique model for studying the macromolecular crowding effect on surface-immobilized enzymes.
KW - biosilica
KW - carbonic anhydrase
KW - enzyme immobilization
KW - Hydrogenovibrio marinus
KW - macromolecular crowding
KW - silica-binding tag
UR - http://www.scopus.com/inward/record.url?scp=85086681799&partnerID=8YFLogxK
U2 - 10.1021/acsami.0c03804
DO - 10.1021/acsami.0c03804
M3 - Article
C2 - 32460480
AN - SCOPUS:85086681799
SN - 1944-8244
VL - 12
SP - 27055
EP - 27063
JO - ACS Applied Materials and Interfaces
JF - ACS Applied Materials and Interfaces
IS - 24
ER -