Marine Biology

, 165:88 | Cite as

Carbonic anhydrase activity in seaweeds: overview and recommendations for measuring activity with an electrometric method, using Macrocystis pyrifera as a model species

  • Pamela A. Fernández
  • Michael Y. Roleda
  • Ralf Rautenberger
  • Catriona L. Hurd
Method

Abstract

Carbonic anhydrase (CA) plays an important physiological role in all biological systems by accelerating the interconversion of CO2 and HCO3. In algae, CA is essential for photosynthesis: external CA (CAext) dehydrates HCO3, enhancing the supply of CO2 to the cell surface, and internal CA (CAint) interconverts HCO3 and CO2 to maintain the inorganic carbon (Ci) pool and supply CO2 to RuBisCO. We first conducted a literature review comparing the conditions in which CA extraction and measurement have been carried out, using the commonly used Wilbur–Anderson method. We found that the assay has been widely modified since its introduction in 1948, mostly without being optimized for the species tested. Based on the review, an optimized protocol for measuring CA in Macrocystis pyrifera was developed, which showed that the assay conditions can strongly affect CA activity. Tris–HCl buffer gave the highest levels of CA activity, but phosphate buffer reduced activity significantly. Buffers containing polyvinylpyrrolidone (PVP) and dithiothreitol (DTT) stabilized CA. Using the optimized assay, CAext and CAint activities were readily measured in Macrocystis with higher precision compared to the non-optimized method. The CAint activity was 2 × higher than CAext, which is attributed to the Ci uptake mechanisms of Macrocystis. This study suggests that the CA assay needs to be optimized for each species prior to experimental work to obtain both accurate and precise results.

Notes

Acknowledgements

We acknowledge the support of a PhD scholarship from the Chilean government to Pamela A. Fernández (BECAS CHILE–CONICYT), a grant from The Royal Society of New Zealand Mardsen fund (UOO0914) to Catriona L. Hurd, and a German Research Foundation grant (RA 2030/1,1) to Ralf Rautenberger. The authors are grateful to Pablo Leal for his help with seaweed collection.

Funding

This study was funded by the Chilean program for BECAS CHILE-CONICYT, for a Royal Society of New Zealand Mardsen grant (UOO0914), and a German Research Foundation grant (RA 2030/1,1).

Compliance with ethical standard

Conflict of interest

Pamela A. Fernández declares that she has no conflict of interest. Michael Y. Roleda declares that he has no conflict of interest. Ralf Rautenberger declares that he has no conflict of interest. Catriona L. Hurd declares that she has no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.

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Authors and Affiliations

  1. 1.Department of BotanyUniversity of OtagoDunedinNew Zealand
  2. 2.Centro i ~ marUniversidad De Los LagosPuerto MonttChile
  3. 3.Norwegian Institute of Bioeconomy ResearchBodøNorway
  4. 4.The Marine Science Institute, College of ScienceUniversity of the Philippines DilimanQuezon CityPhilippines
  5. 5.Institute for Marine and Antarctic Studies (IMAS)University of TasmaniaHobartAustralia

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