- Author
- Hendrik Grootaert (UGent) , Linde Van Landuyt, Paco Hulpiau (UGent) and Nico Callewaert (UGent)
- Organization
- Abstract
- The deoxy sugar L-fucose is frequently found as a glycan constituent on and outside living cells, and in mammals it is involved in a wide range of biological processes including leukocyte trafficking, histo-blood group antigenicity, and antibody effector functions. The manipulation of fucose levels in those biomedically important systems may provide novel insights and therapeutic leads. However, despite the large established sequence diversity of natural fucosidases, so far very few enzymes have been characterized. We explored the diversity of the α-L-fucosidase-containing CAZY family GH29 by bio-informatic analysis, and by the recombinant production and exploration for fucosidase activity of a subset of 82 protein sequences that represent the family’s large sequence diversity. After establishing that most of the corresponding proteins can be readily expressed in E. coli, more than half of the obtained recombinant proteins (57% of the entire subset) showed activity towards the simple chromogenic fucosylated substrate 4-nitrophenyl α-L-fucopyranoside. Thirty-seven of these active GH29 enzymes (and the GH29 subtaxa that they represent) had not been characterized before. With such a sequence diversity-based collection available, it can easily be used to screen for fucosidase activity towards biomedically relevant fucosylated glycoproteins. As an example, the subset was used to screen GH29 members for activity towards the naturally occurring sialyl-Lewis x-type epitope on glycoproteins, and several such enzymes were identified. Together, the results provide a significant increase in the diversity of characterized GH29 enzymes, and the recombinant enzymes constitute a resource for the further functional exploration of this enzyme family.
- Keywords
- Biochemistry, fucosidase, GH29, glycoengineering, glycosyl hydrolase, ALPHA-L-FUCOSIDASES, MULTIPLE SEQUENCE ALIGNMENT, BIFIDOBACTERIUM-BIFIDUM, PROTEIN-STRUCTURE, I-TASSER, FUCOSE, BINDING, 1, 2-ALPHA-L-FUCOSIDASE, OLIGOSACCHARIDES, MECHANISM
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8653532
- MLA
- Grootaert, Hendrik, et al. “Functional Exploration of the GH29 Fucosidase Family.” GLYCOBIOLOGY, vol. 30, no. 9, 2020, pp. 735–45, doi:10.1093/glycob/cwaa023.
- APA
- Grootaert, H., Van Landuyt, L., Hulpiau, P., & Callewaert, N. (2020). Functional exploration of the GH29 fucosidase family. GLYCOBIOLOGY, 30(9), 735–745. https://doi.org/10.1093/glycob/cwaa023
- Chicago author-date
- Grootaert, Hendrik, Linde Van Landuyt, Paco Hulpiau, and Nico Callewaert. 2020. “Functional Exploration of the GH29 Fucosidase Family.” GLYCOBIOLOGY 30 (9): 735–45. https://doi.org/10.1093/glycob/cwaa023.
- Chicago author-date (all authors)
- Grootaert, Hendrik, Linde Van Landuyt, Paco Hulpiau, and Nico Callewaert. 2020. “Functional Exploration of the GH29 Fucosidase Family.” GLYCOBIOLOGY 30 (9): 735–745. doi:10.1093/glycob/cwaa023.
- Vancouver
- 1.Grootaert H, Van Landuyt L, Hulpiau P, Callewaert N. Functional exploration of the GH29 fucosidase family. GLYCOBIOLOGY. 2020;30(9):735–45.
- IEEE
- [1]H. Grootaert, L. Van Landuyt, P. Hulpiau, and N. Callewaert, “Functional exploration of the GH29 fucosidase family,” GLYCOBIOLOGY, vol. 30, no. 9, pp. 735–745, 2020.
@article{8653532,
abstract = {{The deoxy sugar L-fucose is frequently found as a glycan constituent on and outside living cells, and in mammals it is involved in a wide range of biological processes including leukocyte trafficking, histo-blood group antigenicity, and antibody effector functions. The manipulation of fucose levels in those biomedically important systems may provide novel insights and therapeutic leads. However, despite the large established sequence diversity of natural fucosidases, so far very few enzymes have been characterized. We explored the diversity of the α-L-fucosidase-containing CAZY family GH29 by bio-informatic analysis, and by the recombinant production and exploration for fucosidase activity of a subset of 82 protein sequences that represent the family’s large sequence diversity. After establishing that most of the corresponding proteins can be readily expressed in E. coli, more than half of the obtained recombinant proteins (57% of the entire subset) showed activity towards the simple chromogenic fucosylated substrate 4-nitrophenyl α-L-fucopyranoside. Thirty-seven of these active GH29 enzymes (and the GH29 subtaxa that they represent) had not been characterized before. With such a sequence diversity-based collection available, it can easily be used to screen for fucosidase activity towards biomedically relevant fucosylated glycoproteins. As an example, the subset was used to screen GH29 members for activity towards the naturally occurring sialyl-Lewis x-type epitope on glycoproteins, and several such enzymes were identified. Together, the results provide a significant increase in the diversity of characterized GH29 enzymes, and the recombinant enzymes constitute a resource for the further functional exploration of this enzyme family.}},
author = {{Grootaert, Hendrik and Van Landuyt, Linde and Hulpiau, Paco and Callewaert, Nico}},
issn = {{0959-6658}},
journal = {{GLYCOBIOLOGY}},
keywords = {{Biochemistry,fucosidase,GH29,glycoengineering,glycosyl hydrolase,ALPHA-L-FUCOSIDASES,MULTIPLE SEQUENCE ALIGNMENT,BIFIDOBACTERIUM-BIFIDUM,PROTEIN-STRUCTURE,I-TASSER,FUCOSE,BINDING,1,2-ALPHA-L-FUCOSIDASE,OLIGOSACCHARIDES,MECHANISM}},
language = {{eng}},
number = {{9}},
pages = {{735--745}},
title = {{Functional exploration of the GH29 fucosidase family}},
url = {{http://dx.doi.org/10.1093/glycob/cwaa023}},
volume = {{30}},
year = {{2020}},
}
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