scholarly journals Characterization and High-Level Periplasmic Expression of Thermostable α-Carbonic Anhydrase from Thermosulfurimonas Dismutans in Escherichia Coli for CO2 Capture and Utilization

2019 ◽  
Vol 21 (1) ◽  
pp. 103 ◽  
Author(s):  
Byung Hoon Jo ◽  
In Seong Hwang

Carbonic anhydrase (CA) is a diffusion-controlled enzyme that rapidly catalyzes carbon dioxide (CO2) hydration. CA has been considered as a powerful and green catalyst for bioinspired CO2 capture and utilization (CCU). For successful industrial applications, it is necessary to expand the pool of thermostable CAs to meet the stability requirement under various operational conditions. In addition, high-level expression of thermostable CA is desirable for the economical production of the enzyme. In this study, a thermostable CA (tdCA) of Thermosulfurimonas dismutans isolated from a deep-sea hydrothermal vent was expressed in Escherichia coli and characterized in terms of expression level, solubility, activity and stability. tdCA showed higher solubility, activity, and stability compared to those of CA from Thermovibrio ammonificans, one of the most thermostable CAs, under low-salt aqueous conditions. tdCA was engineered for high-level expression by the introduction of a point mutation and periplasmic expression via the Sec-dependent pathway. The combined strategy resulted in a variant showing at least an 8.3-fold higher expression level compared to that of wild-type tdCA. The E. coli cells with the periplasmic tdCA variant were also investigated as an ultra-efficient whole-cell biocatalyst. The engineered bacterium displayed an 11.9-fold higher activity compared to that of the recently reported system with a halophilic CA. Collectively these results demonstrate that the highly expressed periplasmic tdCA variant, either in an isolated form or within a whole-cell platform, is a promising biocatalyst with high activity and stability for CCU applications.

1996 ◽  
Vol 18 (2) ◽  
pp. 163-168 ◽  
Author(s):  
Kwang-Rae Huh ◽  
Eun Hee Cho ◽  
Soo Ok Lee ◽  
Doe Sun Na

2013 ◽  
Vol 41 (3) ◽  
pp. 527-534 ◽  
Author(s):  
Xing-Jun Feng ◽  
Li-Wei Xing ◽  
Di Liu ◽  
Xue-Ying Song ◽  
Chun-Long Liu ◽  
...  

Gene ◽  
1996 ◽  
Vol 176 (1-2) ◽  
pp. 269-272 ◽  
Author(s):  
Bifeng Gao ◽  
Sonia C. Flores ◽  
Swapan K. Bose ◽  
Joe M. McCord

Sign in / Sign up

Export Citation Format

Share Document