138
Views
13
CrossRef citations to date
0
Altmetric
Original Articles

Carbonic anhydrase-based environmental bioassay

Pages 895-903 | Received 08 Oct 2004, Accepted 01 Feb 2005, Published online: 25 Jan 2007
 

Abstract

Carbonic anhydrase (CA) is a metalloenzyme catalysing the reversible idratation of CO2 in H+ and

. It is an ubiquitous enzyme in bacteria, plant and animal kingdoms, playing a fundamental role in a number of physiological processes. Previous studies demonstrated the sensitivity of CA activity to dichlorodiphenyl-dichloroethane (DDT) exposure in birds and to cadmium exposure in teleosts. The aim of the present work was to develop a new in vitro enzymatic bioassay for detecting toxic chemicals in environmental samples as a cost-effective tool in environmental monitoring. This bioassay uses the commercial available CA isozyme II from bovine erythrocytes whose sensitivity to the main classes of chemical pollutants of importance in water quality research was tested in this work. CA activity was determined by a modification of the electrometric method previously described by Wilbur and Anderson [K.M. Wilbur, G.N. Anderson. J. Biol. Chem., 176, 147 (1948).]: briefly, CA activity units were calculated from the rate of H+ production in the reaction mixture (where CO2 was present as substrate) against a blank containing the specific CA inhibitor acetazolamide. [H+] variation was followed at 0°C in the reaction mixture using a Mettler Delta 350 pH-meter. In our experimental set-up bovine CA activity was significantly inhibited by heavy metals (Cd, Cu and Hg), by the organochlorate compound arochlor and by the carbammate pesticides carbaryl in a dose-dependent manner. CA in vitro bioassay can represent a novel tool for rapid and low cost understanding of the toxicity of environmental samples, for assessing bioavailability of pollutants in environmental matrices and their additive or synergistic biological effects when present in mixtures.

Acknowledgements

This study was granted by MEMOBIOMAR project CLUSTER10 − Piano Ambiente Marino (financed by M.I.U.R. and coordinated by CoNISMa).

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.