A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae

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dc.contributor.authorvan Hille, Rob
dc.contributor.authorFagan, Marijke A
dc.contributor.authorBromfield, Lucinda
dc.contributor.authorPott, Robert
dc.date.accessioned2015-04-29T10:53:27Z
dc.date.available2015-04-29T10:53:27Z
dc.date.issued2014-01
dc.description.abstractThe threat of global warming due to CO2 emissions has stimulated research into carbon sequestration and emissions reduction technologies. Alkaline scrubbing allows CO2 to be captured as bicarbonate, which can be photochemically fixed by microalgae. The carbon concentrating mechanism (CCM), of which external carbonic anhydrase is a key component, allows the organisms to utilise this bicarbonate. In order to select a suitable strain for this application, a screening tool is required. The current method for determining carbonic anhydrase activity, the Wilbur and Anderson assay, was found to be unsuitable as a screening tool as the associated error was unacceptably large and tests on whole cells were inconclusive. This paper presents the development of a new, whole cell assay to measure inorganic carbon uptake and external carbonic anhydrase activity, based on classical pH drift experiments. Spirulina platensis was successfully used to develop a correlation between the specific carbon uptake (C) and the specific pH change (dpH). The relationship is described by: C (mmol C (g dry algae)-1 h-1) = 0.064 × (dpH). Inhibitor and salt dissociation tests validated the activity and presence of external carbonic anhydrase, and allowed correlation between the Wilbur and Anderson assay and the new whole cell assay. Screening tests were conducted on Spirulina platensis, Scenedesmus sp., Chlorella vulgaris and Dunaliella salina which were found to have carbon uptake rates of 5.76, 5.86, 3.86 and 2.15 mmol C (g dry algae)-1 h-1 respectively. These results corresponded to the species’ known bicarbonate utilisation abilities and validated the use of the assay as a screening tool.en_ZA
dc.identifier.apacitationvan Hille, R., Fagan, M. A., Bromfield, L., & Pott, R. (2014). A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae. <i>Journal of Applied Phycology</i>, http://hdl.handle.net/11427/12701en_ZA
dc.identifier.chicagocitationvan Hille, Rob, Marijke A Fagan, Lucinda Bromfield, and Robert Pott "A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae." <i>Journal of Applied Phycology</i> (2014) http://hdl.handle.net/11427/12701en_ZA
dc.identifier.citationvan Hille, R., Fagan, M., Bromfield, L. & Pott, R. (2014). A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae. Journal of Applied Phycology, 26(1): 377-385. http://dx.doi.org/10.1007/s10811-013-0076-6en_ZA
dc.identifier.issn9218971en_ZA
dc.identifier.ris TY - Journal Article AU - van Hille, Rob AU - Fagan, Marijke A AU - Bromfield, Lucinda AU - Pott, Robert AB - The threat of global warming due to CO2 emissions has stimulated research into carbon sequestration and emissions reduction technologies. Alkaline scrubbing allows CO2 to be captured as bicarbonate, which can be photochemically fixed by microalgae. The carbon concentrating mechanism (CCM), of which external carbonic anhydrase is a key component, allows the organisms to utilise this bicarbonate. In order to select a suitable strain for this application, a screening tool is required. The current method for determining carbonic anhydrase activity, the Wilbur and Anderson assay, was found to be unsuitable as a screening tool as the associated error was unacceptably large and tests on whole cells were inconclusive. This paper presents the development of a new, whole cell assay to measure inorganic carbon uptake and external carbonic anhydrase activity, based on classical pH drift experiments. Spirulina platensis was successfully used to develop a correlation between the specific carbon uptake (C) and the specific pH change (dpH). The relationship is described by: C (mmol C (g dry algae)-1 h-1) = 0.064 × (dpH). Inhibitor and salt dissociation tests validated the activity and presence of external carbonic anhydrase, and allowed correlation between the Wilbur and Anderson assay and the new whole cell assay. Screening tests were conducted on Spirulina platensis, Scenedesmus sp., Chlorella vulgaris and Dunaliella salina which were found to have carbon uptake rates of 5.76, 5.86, 3.86 and 2.15 mmol C (g dry algae)-1 h-1 respectively. These results corresponded to the species’ known bicarbonate utilisation abilities and validated the use of the assay as a screening tool. DA - 2014-01 DB - OpenUCT DO - 10.1007/s10811-013-0076-6 DP - University of Cape Town J1 - Journal of Applied Phycology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2014 SM - 9218971 T1 - A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae TI - A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae UR - http://hdl.handle.net/11427/12701 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/12701
dc.identifier.urihttp://dx.doi.org/10.1007/s10811-013-0076-6
dc.identifier.vancouvercitationvan Hille R, Fagan MA, Bromfield L, Pott R. A modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgae. Journal of Applied Phycology. 2014; http://hdl.handle.net/11427/12701.en_ZA
dc.languageengen_ZA
dc.publisherSpringeren_ZA
dc.publisher.departmentCentre for Bioprocess Engineering Research
dc.publisher.facultyFaculty of Engineering and the Built Environment
dc.publisher.institutionUniversity of Cape Town
dc.rightsCreative Commons Attribution 4.0 International (CC BY 4.0)*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_ZA
dc.sourceJournal of Applied Phycologyen_ZA
dc.source.urihttp://link.springer.com/journal/10811
dc.titleA modified pH drift assay for inorganic carbon accumulation and external carbonic anhydrase activity in microalgaeen_ZA
dc.typeJournal Articleen_ZA
uct.subject.keywordsaqueous chemistryen_ZA
uct.subject.keywordscarbon concentrating mechanismen_ZA
uct.subject.keywordsscreening testen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourcePost-printen_ZA
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