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Cloning, expression and characterization of the δ‐carbonic anhydrase of Thalassiosira weissflogii (Bacillariophyceae)

Lee, R. B. Y., Smith, J. .A. C. and Rickaby, R. E. M. (2013) Cloning, expression and characterization of the δ‐carbonic anhydrase of Thalassiosira weissflogii (Bacillariophyceae). Journal of Phycology, 49 (1). pp. 170-177. ISSN 1529-8817

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To link to this item DOI: 10.1111/j.1529-8817.2012.01226.x

Abstract/Summary

Carbonic anhydrase (CA) is a ubiquitous metalloenzyme responsible for accelerating the interconversion of CO2 and bicarbonate. Although CAs are involved in a broad range of biochemical processes involving carboxylation or decarboxylation reactions, they are of special interest due to their role in photosynthetic CO2 assimilation in marine phytoplankton, especially under low‐CO2 conditions. Several phylogenetically independent classes of CAs have been identified in a variety of marine phytoplankton. TWCA1, first discovered in Thalassiosira weissflogii (Grunow) G. Fryxell & Hasle, is the founding member of the δ‐class of CAs; these appear to be extracellular enzymes, but are still relatively poorly characterized. To date, it has remained uncertain whether TWCA1 possesses true CA activity due to the difficulty in producing a functional protein in a heterologous expression system. Herein we describe the fusion of a full‐length open reading frame of TWCA1 to the coding sequence of a self‐splicing intein in a pTWIN2 expression vector that has allowed successful production of a functional enzyme in Escherichia coli. Assay of the recombinant protein shows that TWCA1 is a catalytically active δ‐CA possessing both CO2 hydration and esterase activity.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Biological Sciences > Ecology and Evolutionary Biology
ID Code:80106
Publisher:Wiley Online Library

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