Journal article
BGAL1 depletion boosts the level of β-galactosylation of N- and O-glycans in N. benthamiana
- Abstract:
- Glyco-design of proteins is a powerful tool in fundamental studies of structure-function relationship and in obtaining profiles optimized for efficacy of therapeutic glycoproteins. Plants, particularly Nicotiana benthamiana, are attractive hosts to produce recombinant glycoproteins, and recent advances in glyco-engineering facilitate customized N-glycosylation of plant-derived glycoproteins. However, with exception of monoclonal antibodies, homogenous human-like β1,4-galactosylation is very hard to achieve in recombinant glycoproteins. Despite significant efforts to optimize the expression of β1,4-galactosyltransferase, many plant-derived glycoproteins still exhibit incomplete processed N-glycans with heterogeneous terminal galactosylation. The most obvious suspects to be involved in trimming terminal galactose residues are β-galactosidases (BGALs) from the glycosyl hydrolase family GH35. To elucidate the so far uncharacterized mechanisms leading to the trimming of terminal galactose residues from glycans of secreted proteins, we studied a N. benthamiana BGAL known to be active in the apoplast (NbBGAL1). Here, we determined the NbBGAL1 subcellular localization, substrate specificity and in planta biological activity. We show that NbBGAL1 can remove β1,4- and β1,3-galactose residues on both N- and O-glycans. Transient BGAL1 down-regulation by RNA interference (RNAi) and BGAL1 depletion by genome editing drastically reduce β-galactosidase activity in N. benthamiana and increase the amounts of fully galactosylated complex N-glycans on several plant-produced glycoproteins. Altogether, our data demonstrate that NbBGAL1 acts on galactosylated complex N-glycans of plant-produced glycoproteins.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, 3.5MB, Terms of use)
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- Publisher copy:
- 10.1111/pbi.13316
Authors
- Publisher:
- Wiley
- Journal:
- Plant Biotechnology Journal More from this journal
- Volume:
- 18
- Issue:
- 7
- Pages:
- 1537-1549
- Place of publication:
- England
- Publication date:
- 2020-01-11
- Acceptance date:
- 2019-12-05
- DOI:
- EISSN:
-
1467-7652
- ISSN:
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1467-7644
- Pmid:
-
31837192
- Language:
-
English
- Keywords:
- Pubs id:
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1083834
- Local pid:
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pubs:1083834
- Deposit date:
-
2020-03-30
Terms of use
- Copyright holder:
- Kriechbaum et al.
- Copyright date:
- 2020
- Rights statement:
- © 2020 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Licence:
- CC Attribution (CC BY)
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