Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes
| Autor: | Ly Minh Bui, Mateusz Jakub Lezyk, Artem S. Silchenko, Bang H Truong, Jesper Holck, Mikhail I. Kusaykin, Maria Dalgaard Mikkelsen, Hang T T Cao, Anne S. Meyer, Thinh Duc Pham, Van Thi Thanh Tran |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Oligosaccharides Pharmaceutical Science Protein Engineering 01 natural sciences Polymerization Substrate Specificity chemistry.chemical_compound fucoidan Enzyme Stability Drug Discovery Pharmacology Toxicology and Pharmaceutics (miscellaneous) lcsh:QH301-705.5 Sargassum mcclurei chemistry.chemical_classification Gel electrophoresis Endo-fucoidanase biology Sulfates Chemistry Fucoidan Turbinaria ornata Oligosaccharide Recombinant Proteins Biochemistry endo-fucoidanase Glycoside Hydrolases Size-exclusion chromatography Fucus vesiculosus Molecular stabilisation Phaeophyta Flavobacterium Article 03 medical and health sciences Bacterial Proteins Polysaccharides galactofucan molecular stabilisation Enzyme Assays Polysaccharide-Lyases 010405 organic chemistry biology.organism_classification Enzyme assay 0104 chemical sciences 030104 developmental biology Enzyme lcsh:Biology (General) Galactofucan biology.protein |
| Zdroj: | Marine Drugs, Vol 16, Iss 11, p 422 (2018) Marine Drugs Cao, H T T, Mikkelsen, M D, Lezyk, M J, Bui, L M, Tran, V T T, Silchenko, A S, Kusaykin, M I, Pham, T D, Truong, B H, Holck, J & Meyer, A S 2018, ' Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes ', Marine Drugs, vol. 16, no. 11, 422 . https://doi.org/10.3390/md16110422 Volume 16 Issue 11 |
| ISSN: | 1660-3397 |
| Popis: | 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 &alpha (1&rarr 4)-linked l-fucosyls. Fda2 also degraded S. cichorioides and U. pinnatifida fucoidans that have &alpha 3)-linked l-fucosyls in their backbone. In the stabilised form, Fda1 also cleaved &alpha 3) bonds. For the first time, we also show that several enzymes catalyse degradation of S. mcclurei galactofucan-fucoidan, known to contain &alpha 4) and &alpha 3) linked l-fucosyls and galactosyl-&beta 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. |
| Databáze: | OpenAIRE |
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