Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi

Marine coccolithophorid algae are thought to play a significant role in carbon cycling due to their ability to incorporate dissolved inorganic carbon (DIC) into both calcite and photosynthetic products. Among coccolithophorids, Emiliania huxleyi is the most prolific, forming massive blooms that affe...

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Main Authors: Soto, Amelia R., Zheng, Hong, Shoemaker, Dorinda, Rodriguez, Jason, Read, Betsy A., Wahlund, Thomas M.
Formato: Artigo
Idioma:English
Publicado em: American Society for Microbiology 2006
Assuntos:
Genetics and Molecular Biology
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC1538761/
https://ncbi.nlm.nih.gov/pubmed/16885304
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/AEM.00237-06
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spelling pubmed-15387612006-12-01 Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi Soto, Amelia R. Zheng, Hong Shoemaker, Dorinda Rodriguez, Jason Read, Betsy A. Wahlund, Thomas M. Appl Environ Microbiol Genetics and Molecular Biology Marine coccolithophorid algae are thought to play a significant role in carbon cycling due to their ability to incorporate dissolved inorganic carbon (DIC) into both calcite and photosynthetic products. Among coccolithophorids, Emiliania huxleyi is the most prolific, forming massive blooms that affect the global environment. In addition to its ecological importance, the elaborate calcite structures (coccoliths) are being investigated for the design of potential materials for science and biotechnological devices. To date, most of the research focus in this organism has involved the partitioning of DIC between calcification and photosynthesis, primarily using measurements of an external versus internal carbonic anhydrase (CA) activity under defined conditions. The actual genes, proteins, and pathways employed in these processes have not been identified and characterized (see the work of Quinn et al. in this issue [P. Quinn, R. M. Bowers, X. Zhang, T. M. Wahlund, M. A. Fanelli, D. Olszova, and B. A. Read, Appl. Environ. Microbiol. 72:5512-5526, 2006]). In this study, the cloning and preliminary characterization of two genetically distinct carbonic anhydrase cDNAs are described. Phylogenetic analysis indicated that these two genes belonged to the gamma (γ-EhCA2) and delta (δ-EhCA1) classes of carbonic anhydrases. The deduced amino acid sequence of δ-EhCA1 revealed that it encodes a protein of 702 amino acids (aa) (ca. 77.3 kDa), with a transmembrane N-terminal region of 373 aa and an in-frame C-terminal open reading frame of 329 aa that defines the CA region. The γ-EhCA2 protein was 235 aa in length (ca. 24.9 kDa) and was successfully expressed in Escherichia coli BL21(DE3) and purified as an active recombinant CA. The expression levels of each transcript from quantitative reverse transcription-PCR experiments under bicarbonate limitation and over a 24-h time course suggest that these isozymes perform different functions in E. huxleyi. American Society for Microbiology 2006-08 /pmc/articles/PMC1538761/ /pubmed/16885304 http://dx.doi.org/10.1128/AEM.00237-06 Text en Copyright © 2006, American Society for Microbiology
institution US National Library of Medicine
collection PubMed Central
language English
format Article
topic Genetics and Molecular Biology
spellingShingle Genetics and Molecular Biology
Soto, Amelia R.
Zheng, Hong
Shoemaker, Dorinda
Rodriguez, Jason
Read, Betsy A.
Wahlund, Thomas M.
Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
description Marine coccolithophorid algae are thought to play a significant role in carbon cycling due to their ability to incorporate dissolved inorganic carbon (DIC) into both calcite and photosynthetic products. Among coccolithophorids, Emiliania huxleyi is the most prolific, forming massive blooms that affect the global environment. In addition to its ecological importance, the elaborate calcite structures (coccoliths) are being investigated for the design of potential materials for science and biotechnological devices. To date, most of the research focus in this organism has involved the partitioning of DIC between calcification and photosynthesis, primarily using measurements of an external versus internal carbonic anhydrase (CA) activity under defined conditions. The actual genes, proteins, and pathways employed in these processes have not been identified and characterized (see the work of Quinn et al. in this issue [P. Quinn, R. M. Bowers, X. Zhang, T. M. Wahlund, M. A. Fanelli, D. Olszova, and B. A. Read, Appl. Environ. Microbiol. 72:5512-5526, 2006]). In this study, the cloning and preliminary characterization of two genetically distinct carbonic anhydrase cDNAs are described. Phylogenetic analysis indicated that these two genes belonged to the gamma (γ-EhCA2) and delta (δ-EhCA1) classes of carbonic anhydrases. The deduced amino acid sequence of δ-EhCA1 revealed that it encodes a protein of 702 amino acids (aa) (ca. 77.3 kDa), with a transmembrane N-terminal region of 373 aa and an in-frame C-terminal open reading frame of 329 aa that defines the CA region. The γ-EhCA2 protein was 235 aa in length (ca. 24.9 kDa) and was successfully expressed in Escherichia coli BL21(DE3) and purified as an active recombinant CA. The expression levels of each transcript from quantitative reverse transcription-PCR experiments under bicarbonate limitation and over a 24-h time course suggest that these isozymes perform different functions in E. huxleyi.
author Soto, Amelia R.
Zheng, Hong
Shoemaker, Dorinda
Rodriguez, Jason
Read, Betsy A.
Wahlund, Thomas M.
author_facet Soto, Amelia R.
Zheng, Hong
Shoemaker, Dorinda
Rodriguez, Jason
Read, Betsy A.
Wahlund, Thomas M.
author_sort Soto, Amelia R.
title Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
title_short Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
title_full Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
title_fullStr Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
title_full_unstemmed Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi
title_sort identification and preliminary characterization of two cdnas encoding unique carbonic anhydrases from the marine alga emiliania huxleyi
publisher American Society for Microbiology
publisher_facet American Society for Microbiology
publishDate 2006
url https://ncbi.nlm.nih.gov/pmc/articles/PMC1538761/
https://ncbi.nlm.nih.gov/pubmed/16885304
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/AEM.00237-06
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