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The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans
Fucoidans are complex bioactive sulfated fucosyl-polysaccharides primarily found in brown macroalgae. Endo-fucoidanases catalyze the specific hydrolysis of α-L-fucosyl linkages in fucoidans and can be utilized to tailor-make fucoidan oligosaccharides and elucidate new structural details of fucoidans...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9147238/ https://www.ncbi.nlm.nih.gov/pubmed/35621956 http://dx.doi.org/10.3390/md20050305 |
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author | Tran, Vy Ha Nguyen Nguyen, Thuan Thi Meier, Sebastian Holck, Jesper Cao, Hang Thi Thuy Van, Tran Thi Thanh Meyer, Anne S. Mikkelsen, Maria Dalgaard |
author_facet | Tran, Vy Ha Nguyen Nguyen, Thuan Thi Meier, Sebastian Holck, Jesper Cao, Hang Thi Thuy Van, Tran Thi Thanh Meyer, Anne S. Mikkelsen, Maria Dalgaard |
author_sort | Tran, Vy Ha Nguyen |
collection | PubMed |
description | Fucoidans are complex bioactive sulfated fucosyl-polysaccharides primarily found in brown macroalgae. Endo-fucoidanases catalyze the specific hydrolysis of α-L-fucosyl linkages in fucoidans and can be utilized to tailor-make fucoidan oligosaccharides and elucidate new structural details of fucoidans. In this study, an endo-α(1,3)-fucoidanase encoding gene, Mef2, from the marine bacterium Muricauda eckloniae, was cloned, and the Mef2 protein was functionally characterized. Based on the primary sequence, Mef2 was suggested to belong to the glycosyl hydrolase family 107 (GH107) in the Carbohydrate Active enZyme database (CAZy). The Mef2 fucoidanase showed maximal activity at pH 8 and 35 °C, although it could tolerate temperatures up to 50 °C. Ca(2+) was shown to increase the melting temperature from 38 to 44 °C and was furthermore required for optimal activity of Mef2. The substrate specificity of Mef2 was investigated, and Fourier transform infrared spectroscopy (FTIR) was used to determine the enzymatic activity (Units per μM enzyme: U(f)/μM) of Mef2 on two structurally different fucoidans, showing an activity of 1.2 × 10(−3) U(f)/μM and 3.6 × 10(−3) U(f)/μM on fucoidans from Fucus evanescens and Saccharina latissima, respectively. Interestingly, Mef2 was identified as the first described fucoidanase active on fucoidans from S. latissima. The fucoidan oligosaccharides released by Mef2 consisted of a backbone of α(1,3)-linked fucosyl residues with unique and novel α(1,4)-linked fucosyl branches, not previously identified in fucoidans from S. latissima. |
format | Online Article Text |
id | pubmed-9147238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-91472382022-05-29 The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans Tran, Vy Ha Nguyen Nguyen, Thuan Thi Meier, Sebastian Holck, Jesper Cao, Hang Thi Thuy Van, Tran Thi Thanh Meyer, Anne S. Mikkelsen, Maria Dalgaard Mar Drugs Article Fucoidans are complex bioactive sulfated fucosyl-polysaccharides primarily found in brown macroalgae. Endo-fucoidanases catalyze the specific hydrolysis of α-L-fucosyl linkages in fucoidans and can be utilized to tailor-make fucoidan oligosaccharides and elucidate new structural details of fucoidans. In this study, an endo-α(1,3)-fucoidanase encoding gene, Mef2, from the marine bacterium Muricauda eckloniae, was cloned, and the Mef2 protein was functionally characterized. Based on the primary sequence, Mef2 was suggested to belong to the glycosyl hydrolase family 107 (GH107) in the Carbohydrate Active enZyme database (CAZy). The Mef2 fucoidanase showed maximal activity at pH 8 and 35 °C, although it could tolerate temperatures up to 50 °C. Ca(2+) was shown to increase the melting temperature from 38 to 44 °C and was furthermore required for optimal activity of Mef2. The substrate specificity of Mef2 was investigated, and Fourier transform infrared spectroscopy (FTIR) was used to determine the enzymatic activity (Units per μM enzyme: U(f)/μM) of Mef2 on two structurally different fucoidans, showing an activity of 1.2 × 10(−3) U(f)/μM and 3.6 × 10(−3) U(f)/μM on fucoidans from Fucus evanescens and Saccharina latissima, respectively. Interestingly, Mef2 was identified as the first described fucoidanase active on fucoidans from S. latissima. The fucoidan oligosaccharides released by Mef2 consisted of a backbone of α(1,3)-linked fucosyl residues with unique and novel α(1,4)-linked fucosyl branches, not previously identified in fucoidans from S. latissima. MDPI 2022-04-29 /pmc/articles/PMC9147238/ /pubmed/35621956 http://dx.doi.org/10.3390/md20050305 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Tran, Vy Ha Nguyen Nguyen, Thuan Thi Meier, Sebastian Holck, Jesper Cao, Hang Thi Thuy Van, Tran Thi Thanh Meyer, Anne S. Mikkelsen, Maria Dalgaard The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title | The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title_full | The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title_fullStr | The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title_full_unstemmed | The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title_short | The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans |
title_sort | endo-α(1,3)-fucoidanase mef2 releases uniquely branched oligosaccharides from saccharina latissima fucoidans |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9147238/ https://www.ncbi.nlm.nih.gov/pubmed/35621956 http://dx.doi.org/10.3390/md20050305 |
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