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Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology

The carrageenophyte red alga Chondrus crispus produces three family 16 glycoside hydrolases (CcGH16-1, CcGH16-2, and CcGH16-3). Phylogenetically, the red algal GH16 members are closely related to bacterial GH16 homologs from subfamilies 13 and 14, which have characterized marine bacterial β-carragee...

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Autores principales: Manat, Guillaume, Fanuel, Mathieu, Jouanneau, Diane, Jam, Murielle, Mac-Bear, Jessica, Rogniaux, Hélène, Mora, Théo, Larocque, Robert, Lipinska, Agnieszka, Czjzek, Mirjam, Ropartz, David, Ficko-Blean, Elizabeth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9771727/
https://www.ncbi.nlm.nih.gov/pubmed/36402445
http://dx.doi.org/10.1016/j.jbc.2022.102707
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author Manat, Guillaume
Fanuel, Mathieu
Jouanneau, Diane
Jam, Murielle
Mac-Bear, Jessica
Rogniaux, Hélène
Mora, Théo
Larocque, Robert
Lipinska, Agnieszka
Czjzek, Mirjam
Ropartz, David
Ficko-Blean, Elizabeth
author_facet Manat, Guillaume
Fanuel, Mathieu
Jouanneau, Diane
Jam, Murielle
Mac-Bear, Jessica
Rogniaux, Hélène
Mora, Théo
Larocque, Robert
Lipinska, Agnieszka
Czjzek, Mirjam
Ropartz, David
Ficko-Blean, Elizabeth
author_sort Manat, Guillaume
collection PubMed
description The carrageenophyte red alga Chondrus crispus produces three family 16 glycoside hydrolases (CcGH16-1, CcGH16-2, and CcGH16-3). Phylogenetically, the red algal GH16 members are closely related to bacterial GH16 homologs from subfamilies 13 and 14, which have characterized marine bacterial β-carrageenase and β-porphyranase activities, respectively, yet the functions of these CcGH16 hydrolases have not been determined. Here, we first confirmed the gene locus of the ccgh16-3 gene in the alga to facilitate further investigation. Next, our biochemical characterization of CcGH16-3 revealed an unexpected β-porphyranase activity, since porphyran is not a known component of the C. crispus extracellular matrix. Kinetic characterization was undertaken on natural porphyran substrate with an experimentally determined molecular weight. We found CcGH16-3 has a pH optimum between 7.5 and 8.0; however, it exhibits reasonably stable activity over a large pH range (pH 7.0–9.0). CcGH16-3 has a K(M) of 4.0 ± 0.8 μM, a k(cat) of 79.9 ± 6.9 s(−1), and a k(cat)/K(M) of 20.1 ± 1.7 μM(−1) s(−1). We structurally examined fine enzymatic specificity by performing a subsite dissection. CcGH16-3 has a strict requirement for D-galactose and L-galactose-6-sulfate in its −1 and +1 subsites, respectively, whereas the outer subsites are less restrictive. CcGH16-3 is one of a handful of algal enzymes characterized with a specificity for a polysaccharide unknown to be found in their own extracellular matrix. This β-porphyranase activity in a carrageenophyte red alga may provide defense against red algal pathogens or provide a competitive advantage in niche colonization.
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spelling pubmed-97717272022-12-23 Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology Manat, Guillaume Fanuel, Mathieu Jouanneau, Diane Jam, Murielle Mac-Bear, Jessica Rogniaux, Hélène Mora, Théo Larocque, Robert Lipinska, Agnieszka Czjzek, Mirjam Ropartz, David Ficko-Blean, Elizabeth J Biol Chem Research Article The carrageenophyte red alga Chondrus crispus produces three family 16 glycoside hydrolases (CcGH16-1, CcGH16-2, and CcGH16-3). Phylogenetically, the red algal GH16 members are closely related to bacterial GH16 homologs from subfamilies 13 and 14, which have characterized marine bacterial β-carrageenase and β-porphyranase activities, respectively, yet the functions of these CcGH16 hydrolases have not been determined. Here, we first confirmed the gene locus of the ccgh16-3 gene in the alga to facilitate further investigation. Next, our biochemical characterization of CcGH16-3 revealed an unexpected β-porphyranase activity, since porphyran is not a known component of the C. crispus extracellular matrix. Kinetic characterization was undertaken on natural porphyran substrate with an experimentally determined molecular weight. We found CcGH16-3 has a pH optimum between 7.5 and 8.0; however, it exhibits reasonably stable activity over a large pH range (pH 7.0–9.0). CcGH16-3 has a K(M) of 4.0 ± 0.8 μM, a k(cat) of 79.9 ± 6.9 s(−1), and a k(cat)/K(M) of 20.1 ± 1.7 μM(−1) s(−1). We structurally examined fine enzymatic specificity by performing a subsite dissection. CcGH16-3 has a strict requirement for D-galactose and L-galactose-6-sulfate in its −1 and +1 subsites, respectively, whereas the outer subsites are less restrictive. CcGH16-3 is one of a handful of algal enzymes characterized with a specificity for a polysaccharide unknown to be found in their own extracellular matrix. This β-porphyranase activity in a carrageenophyte red alga may provide defense against red algal pathogens or provide a competitive advantage in niche colonization. American Society for Biochemistry and Molecular Biology 2022-11-17 /pmc/articles/PMC9771727/ /pubmed/36402445 http://dx.doi.org/10.1016/j.jbc.2022.102707 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Manat, Guillaume
Fanuel, Mathieu
Jouanneau, Diane
Jam, Murielle
Mac-Bear, Jessica
Rogniaux, Hélène
Mora, Théo
Larocque, Robert
Lipinska, Agnieszka
Czjzek, Mirjam
Ropartz, David
Ficko-Blean, Elizabeth
Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title_full Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title_fullStr Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title_full_unstemmed Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title_short Specificity of a β-porphyranase produced by the carrageenophyte red alga Chondrus crispus and implications of this unexpected activity on red algal biology
title_sort specificity of a β-porphyranase produced by the carrageenophyte red alga chondrus crispus and implications of this unexpected activity on red algal biology
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9771727/
https://www.ncbi.nlm.nih.gov/pubmed/36402445
http://dx.doi.org/10.1016/j.jbc.2022.102707
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