Cargando…

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: Sargassum mcclurei, Fu...

Descripción completa

Detalles Bibliográficos
Autores principales: Cao, Hang T. T., Mikkelsen, Maria D., Lezyk, Mateusz J., Bui, Ly M., Tran, Van T. T., Silchenko, Artem S., Kusaykin, Mikhail I., Pham, Thinh D., Truong, Bang H., Holck, Jesper, Meyer, Anne S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267234/
https://www.ncbi.nlm.nih.gov/pubmed/30388774
http://dx.doi.org/10.3390/md16110422
_version_ 1783376020738408448
author Cao, Hang T. T.
Mikkelsen, Maria D.
Lezyk, Mateusz J.
Bui, Ly M.
Tran, Van T. T.
Silchenko, Artem S.
Kusaykin, Mikhail I.
Pham, Thinh D.
Truong, Bang H.
Holck, Jesper
Meyer, Anne S.
author_facet Cao, Hang T. T.
Mikkelsen, Maria D.
Lezyk, Mateusz J.
Bui, Ly M.
Tran, Van T. T.
Silchenko, Artem S.
Kusaykin, Mikhail I.
Pham, Thinh D.
Truong, Bang H.
Holck, Jesper
Meyer, Anne S.
author_sort Cao, Hang T. T.
collection PubMed
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: Sargassum mcclurei, Fucus evanescens, Fucus vesiculosus, Turbinaria ornata, Saccharina cichorioides, and Undaria pinnatifida. 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 l-fucosyls. Fda2 also degraded S. cichorioides and U. pinnatifida fucoidans that have α(1→3)-linked l-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 S. mcclurei galactofucan-fucoidan, known to contain α(1→4) and α(1→3) linked l-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.
format Online
Article
Text
id pubmed-6267234
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-62672342018-12-06 Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes Cao, Hang T. T. Mikkelsen, Maria D. Lezyk, Mateusz J. Bui, Ly M. Tran, Van T. T. Silchenko, Artem S. Kusaykin, Mikhail I. Pham, Thinh D. Truong, Bang H. Holck, Jesper Meyer, Anne S. Mar Drugs Article 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: Sargassum mcclurei, Fucus evanescens, Fucus vesiculosus, Turbinaria ornata, Saccharina cichorioides, and Undaria pinnatifida. 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 l-fucosyls. Fda2 also degraded S. cichorioides and U. pinnatifida fucoidans that have α(1→3)-linked l-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 S. mcclurei galactofucan-fucoidan, known to contain α(1→4) and α(1→3) linked l-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. MDPI 2018-11-01 /pmc/articles/PMC6267234/ /pubmed/30388774 http://dx.doi.org/10.3390/md16110422 Text en © 2018 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cao, Hang T. T.
Mikkelsen, Maria D.
Lezyk, Mateusz J.
Bui, Ly M.
Tran, Van T. T.
Silchenko, Artem S.
Kusaykin, Mikhail I.
Pham, Thinh D.
Truong, Bang H.
Holck, Jesper
Meyer, Anne S.
Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267234/
https://www.ncbi.nlm.nih.gov/pubmed/30388774
http://dx.doi.org/10.3390/md16110422
work_keys_str_mv AT caohangtt novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT mikkelsenmariad novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT lezykmateuszj novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT builym novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT tranvantt novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT silchenkoartems novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT kusaykinmikhaili novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT phamthinhd novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT truongbangh novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT holckjesper novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes
AT meyerannes novelenzymeactionsforsulphatedgalactofucandepolymerisationandanewengineeringstrategyformolecularstabilisationoffucoidandegradingenzymes