Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes
Fucoidans from brown macroalgae have beneficial biomedical properties but their use as pharma products requires homogenous oligomeric products. In this study, the action of five recombinant microbial fucoidan degrading enzymes were evaluated on fucoidans from brown macroalgae: <i>Sargassum mcc...
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MDPI AG
2018-11-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/16/11/422 |
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| author | Hang T. T. Cao Maria D. Mikkelsen Mateusz J. Lezyk Ly M. Bui Van T. T. Tran Artem S. Silchenko Mikhail I. Kusaykin Thinh D. Pham Bang H. Truong Jesper Holck Anne S. Meyer |
| author_facet | Hang T. T. Cao Maria D. Mikkelsen Mateusz J. Lezyk Ly M. Bui Van T. T. Tran Artem S. Silchenko Mikhail I. Kusaykin Thinh D. Pham Bang H. Truong Jesper Holck Anne S. Meyer |
| author_sort | Hang T. T. Cao |
| collection | DOAJ |
| description | Fucoidans from brown macroalgae have beneficial biomedical properties but their use as pharma products requires homogenous oligomeric products. In this study, the action of five recombinant microbial fucoidan degrading enzymes were evaluated on fucoidans from brown macroalgae: <i>Sargassum mcclurei</i>, <i>Fucus evanescens</i>, <i>Fucus vesiculosus</i>, <i>Turbinaria ornata</i>, <i>Saccharina cichorioides</i>, and <i>Undaria pinnatifida</i>. The enzymes included three endo-fucoidanases (EC 3.2.1.-GH 107), FcnA2, Fda1, and Fda2, and two unclassified endo-fucoglucuronomannan lyases, FdlA and FdlB. The oligosaccharide product profiles were assessed by carbohydrate-polyacrylamide gel electrophoresis and size exclusion chromatography. The recombinant enzymes FcnA2, Fda1, and Fda2 were unstable but were stabilised by truncation of the C-terminal end (removing up to 40% of the enzyme sequence). All five enzymes catalysed degradation of fucoidans containing α(1→4)-linked <span style="font-variant: small-caps;">l</span>-fucosyls. Fda2 also degraded <i>S. cichorioides</i> and <i>U. pinnatifida</i> fucoidans that have α(1→3)-linked <span style="font-variant: small-caps;">l</span>-fucosyls in their backbone. In the stabilised form, Fda1 also cleaved α(1→3) bonds. For the first time, we also show that several enzymes catalyse degradation of <i>S. mcclurei</i> galactofucan-fucoidan, known to contain α(1→4) and α(1→3) linked <span style="font-variant: small-caps;">l</span>-fucosyls and galactosyl-β(1→3) bonds in the backbone. These data enhance our understanding of fucoidan degrading enzymes and their substrate preferences and may assist development of enzyme-assisted production of defined fuco-oligosaccharides from fucoidan substrates. |
| first_indexed | 2024-04-11T18:03:20Z |
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| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-11T18:03:20Z |
| publishDate | 2018-11-01 |
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| series | Marine Drugs |
| spelling | doaj.art-acd25ef4b3b74d3aa283586f90c3a65e2022-12-22T04:10:24ZengMDPI AGMarine Drugs1660-33972018-11-01161142210.3390/md16110422md16110422Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading EnzymesHang T. T. Cao0Maria D. Mikkelsen1Mateusz J. Lezyk2Ly M. Bui3Van T. T. Tran4Artem S. Silchenko5Mikhail I. Kusaykin6Thinh D. Pham7Bang H. Truong8Jesper Holck9Anne S. Meyer10Protein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, DenmarkProtein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, DenmarkProtein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, DenmarkNhaTrang Institute of Technology Research and Application, Vietnam Academy of Science and Technology, 02 Hung Vuong Street, Nhatrang 650000, VietnamNhaTrang Institute of Technology Research and Application, Vietnam Academy of Science and Technology, 02 Hung Vuong Street, Nhatrang 650000, VietnamLaboratory of Enzyme Chemistry, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 159 100-Let Vladivostoku Ave., Vladivostok 690022, RussiaLaboratory of Enzyme Chemistry, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 159 100-Let Vladivostoku Ave., Vladivostok 690022, RussiaNhaTrang Institute of Technology Research and Application, Vietnam Academy of Science and Technology, 02 Hung Vuong Street, Nhatrang 650000, VietnamNhaTrang Institute of Technology Research and Application, Vietnam Academy of Science and Technology, 02 Hung Vuong Street, Nhatrang 650000, VietnamProtein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, DenmarkProtein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, DenmarkFucoidans from brown macroalgae have beneficial biomedical properties but their use as pharma products requires homogenous oligomeric products. In this study, the action of five recombinant microbial fucoidan degrading enzymes were evaluated on fucoidans from brown macroalgae: <i>Sargassum mcclurei</i>, <i>Fucus evanescens</i>, <i>Fucus vesiculosus</i>, <i>Turbinaria ornata</i>, <i>Saccharina cichorioides</i>, and <i>Undaria pinnatifida</i>. The enzymes included three endo-fucoidanases (EC 3.2.1.-GH 107), FcnA2, Fda1, and Fda2, and two unclassified endo-fucoglucuronomannan lyases, FdlA and FdlB. The oligosaccharide product profiles were assessed by carbohydrate-polyacrylamide gel electrophoresis and size exclusion chromatography. The recombinant enzymes FcnA2, Fda1, and Fda2 were unstable but were stabilised by truncation of the C-terminal end (removing up to 40% of the enzyme sequence). All five enzymes catalysed degradation of fucoidans containing α(1→4)-linked <span style="font-variant: small-caps;">l</span>-fucosyls. Fda2 also degraded <i>S. cichorioides</i> and <i>U. pinnatifida</i> fucoidans that have α(1→3)-linked <span style="font-variant: small-caps;">l</span>-fucosyls in their backbone. In the stabilised form, Fda1 also cleaved α(1→3) bonds. For the first time, we also show that several enzymes catalyse degradation of <i>S. mcclurei</i> galactofucan-fucoidan, known to contain α(1→4) and α(1→3) linked <span style="font-variant: small-caps;">l</span>-fucosyls and galactosyl-β(1→3) bonds in the backbone. These data enhance our understanding of fucoidan degrading enzymes and their substrate preferences and may assist development of enzyme-assisted production of defined fuco-oligosaccharides from fucoidan substrates.https://www.mdpi.com/1660-3397/16/11/422fucoidanendo-fucoidanasegalactofucanmolecular stabilisation<i>Sargassum mcclurei</i><i>Turbinaria ornata</i> |
| spellingShingle | Hang T. T. Cao Maria D. Mikkelsen Mateusz J. Lezyk Ly M. Bui Van T. T. Tran Artem S. Silchenko Mikhail I. Kusaykin Thinh D. Pham Bang H. Truong Jesper Holck Anne S. Meyer Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes Marine Drugs fucoidan endo-fucoidanase galactofucan molecular stabilisation <i>Sargassum mcclurei</i> <i>Turbinaria ornata</i> |
| title | Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes |
| title_full | Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes |
| title_fullStr | Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes |
| title_full_unstemmed | Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes |
| title_short | Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes |
| title_sort | novel enzyme actions for sulphated galactofucan depolymerisation and a new engineering strategy for molecular stabilisation of fucoidan degrading enzymes |
| topic | fucoidan endo-fucoidanase galactofucan molecular stabilisation <i>Sargassum mcclurei</i> <i>Turbinaria ornata</i> |
| url | https://www.mdpi.com/1660-3397/16/11/422 |
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