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.

doi:10.3390/md16110422

Open AccessFeature PaperArticle

Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes

by

Hang T. T. Cao

^1,2

,

Maria D. Mikkelsen

¹,

Mateusz J. Lezyk

¹,

Ly M. Bui

²,

Van T. T. Tran

²,

Artem S. Silchenko

³,

Mikhail I. Kusaykin

³

,

Thinh D. Pham

²

,

Bang H. Truong

²,

Jesper Holck

¹

and

Anne S. Meyer

^1,*

¹

Protein Chemistry and Enzyme Technology, DTU Bioengineering, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 221, 2800 Kongens Lyngby, Denmark

²

NhaTrang Institute of Technology Research and Application, Vietnam Academy of Science and Technology, 02 Hung Vuong Street, Nhatrang 650000, Vietnam

³

Laboratory 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, Russia

^*

Author to whom correspondence should be addressed.

Mar. Drugs 2018, 16(11), 422; https://doi.org/10.3390/md16110422

Submission received: 30 September 2018 / Revised: 19 October 2018 / Accepted: 22 October 2018 / Published: 1 November 2018

(This article belongs to the Special Issue Marine Enzymes: Sources, Biochemistry and Bioprocesses for Marine Biotechnology)

Download

Browse Figures

Versions Notes

Abstract

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.

Keywords: fucoidan; endo-fucoidanase; galactofucan; molecular stabilisation; Sargassum mcclurei; Turbinaria ornata

Share and Cite

MDPI and ACS Style

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.; et al. Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes. Mar. Drugs 2018, 16, 422. https://doi.org/10.3390/md16110422

AMA Style

Cao HTT, Mikkelsen MD, Lezyk MJ, Bui LM, Tran VTT, Silchenko AS, Kusaykin MI, Pham TD, Truong BH, Holck J, et al. Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes. Marine Drugs. 2018; 16(11):422. https://doi.org/10.3390/md16110422

Chicago/Turabian Style

Cao, Hang T. T., 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, and et al. 2018. "Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes" Marine Drugs 16, no. 11: 422. https://doi.org/10.3390/md16110422

APA Style

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, 16(11), 422. https://doi.org/10.3390/md16110422

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Menu

Novel Enzyme Actions for Sulphated Galactofucan Depolymerisation and a New Engineering Strategy for Molecular Stabilisation of Fucoidan Degrading Enzymes

Abstract

Share and Cite

Article Metrics

Article Access Statistics

Further Information

Guidelines

MDPI Initiatives

Follow MDPI