Increased expression level and catalytic activity of internally-duplicated carbonic anhydrase from Dunaliella species by reconstitution of two separate domains

2012 ◽  
Vol 47 (9) ◽  
pp. 1423-1427 ◽  
Author(s):  
Mi-Ran Ki ◽  
Bashistha Kumar Kanth ◽  
Ki Ha Min ◽  
Jinwon Lee ◽  
Seung Pil Pack
Keyword(s):  
Catalytic Activity ◽  
Carbonic Anhydrase ◽  
Expression Level ◽  
Dunaliella Species

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
Keyword(s):  
Escherichia Coli ◽  
Carbonic Anhydrase ◽  
Co2 Capture ◽  
Industrial Applications ◽  
Expression Level ◽  
High Level Expression ◽  
Whole Cell ◽  
Operational Conditions ◽  
Periplasmic Expression ◽  
High Level

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.

scholarly journals Azobenzene-based inhibitors of human carbonic anhydrase II

2015 ◽  
Vol 11 ◽  
pp. 1129-1135 ◽  
Author(s):  
Leander Simon Runtsch ◽  
David Michael Barber ◽  
Peter Mayer ◽  
Michael Groll ◽  
Dirk Trauner ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Catalytic Activity ◽  
Carbonic Anhydrase ◽  
Drug Class ◽  
Carbonic Anhydrase Ii ◽  
Carbonic Anhydrases ◽  
X Ray ◽  
X Ray Crystallography ◽  
Human Carbonic Anhydrase

Aryl sulfonamides are a widely used drug class for the inhibition of carbonic anhydrases. In the context of our program of photochromic pharmacophores we were interested in the exploration of azobenzene-containing sulfonamides to block the catalytic activity of human carbonic anhydrase II (hCAII). Herein, we report the synthesis and in vitro evaluation of a small library of nine photochromic sulfonamides towards hCAII. All molecules are azobenzene-4-sulfonamides, which are substituted by different functional groups in the 4´-position and were characterized by X-ray crystallography. We aimed to investigate the influence of electron-donating or electron-withdrawing substituents on the inhibitory constant K i. With the aid of an hCAII crystal structure bound to one of the synthesized azobenzenes, we found that the electronic structure does not strongly affect inhibition. Taken together, all compounds are strong blockers of hCAII with K i = 25–65 nM that are potentially photochromic and thus combine studies from chemical synthesis, crystallography and enzyme kinetics.

In vitro functional reconstitution of duplicate alpha-type carbonic anhydrase of Dunaliella species from purified individual recombinant N and C-half domains

2012 ◽  
Vol 29 ◽  
pp. S97
Author(s):  
Mi-Ran Ki ◽  
Bashistha Kumar Kanth ◽  
Ki Ha Min ◽  
Eui Kyoung Jang ◽  
Ki Baek Yeo ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Dunaliella Species ◽  
Alpha Type

scholarly journals Carbonic anhydrase VII is S-glutathionylated without loss of catalytic activity and affinity for sulfonamide inhibitors

2012 ◽  
Vol 22 (4) ◽  
pp. 1560-1564 ◽  
Author(s):  
Emanuela Truppo ◽  
Claudiu T. Supuran ◽  
Annamaria Sandomenico ◽  
Daniela Vullo ◽  
Alessio Innocenti ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Carbonic Anhydrase

Catalytic activity of a ζ-class zinc and cadmium containing carbonic anhydrase. Compared work mechanisms

2011 ◽  
Vol 13 (8) ◽  
pp. 3468 ◽  
Author(s):  
Orazio Amata ◽  
Tiziana Marino ◽  
Nino Russo ◽  
Marirosa Toscano
Keyword(s):  
Catalytic Activity ◽  
Carbonic Anhydrase

Highly active enzymes immobilized in large pore colloidal mesoporous silica nanoparticles

2019 ◽  
Vol 43 (4) ◽  
pp. 1671-1680 ◽  
Author(s):  
Dorothée Gößl ◽  
Helena Singer ◽  
Hsin-Yi Chiu ◽  
Alexandra Schmidt ◽  
Martina Lichtnecker ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Carbonic Anhydrase ◽  
Horseradish Peroxidase ◽  
Mesoporous Silica ◽  
Silica Nanoparticles ◽  
Mesoporous Silica Nanoparticles ◽  
Click Reactions ◽  
Highly Active ◽  
Large Pore ◽  
Colorimetric Assays

Carbonic anhydrase and horseradish peroxidase are immobilized inside the ordered material by click reactions. Colorimetric assays prove their catalytic activity.

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