Journal article
Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes
Enzyme Technology, Section for Protein Chemistry and Enzyme Technology, Department of Biotechnology and Biomedicine, Technical University of Denmark1
Section for Protein Chemistry and Enzyme Technology, Department of Biotechnology and Biomedicine, Technical University of Denmark2
Department of Biotechnology and Biomedicine, Technical University of Denmark3
Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark4
Department of Chemical and Biochemical Engineering, Technical University of Denmark5
Vietnamese Academy of Science and Technology6
Russian Academy of Sciences7
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.
Language: | English |
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Publisher: | MDPI |
Year: | 2018 |
Pages: | 422 |
ISSN: | 16603397 |
Types: | Journal article |
DOI: | 10.3390/md16110422 |
ORCIDs: | 0000-0001-7117-8103 , 0000-0002-8863-7385 , Holck, Jesper and Meyer, Anne S. |
Endo-fucoidanase Fucoidan Galactofucan Molecular stabilisation Sargassum mcclurei Turbinaria ornata
<i>Sargassum mcclurei</i> <i>Turbinaria ornata</i> Bacterial Proteins Biology (General) Enzyme Assays Enzyme Stability Flavobacterium Glycoside Hydrolases Oligosaccharides Phaeophyceae Polymerization Polysaccharide-Lyases Polysaccharides Protein Engineering QH301-705.5 Recombinant Proteins Substrate Specificity Sulfates endo-fucoidanase fucoidan galactofucan molecular stabilisation