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Exploring a novel β-1,3-glucanosyltransglycosylase, MlGH17B, from a marine Muricauda lutaonensis strain for modification of laminari-oligosaccharides

Allahgholi, Leila LU ; Derks, Maik G.N. ; Dobruchowska, Justyna M ; Jasilionis, Andrius LU ; Moenaert, Antoine ; Jouy, Léonie ; Gulshan Kazi, Zubaida LU ; Linares-Pastén, Javier A LU orcid ; Friðjónsson, Ólafur H and Hreggviðsson, Guðmundur Óli , et al. (2024) In Glycobiology
Abstract

The marine environment, contains plentiful renewable resources, e.g. macroalgae with unique polysaccharides, motivating search for enzymes from marine microorganisms to explore conversion possibilities of the polysaccharides. In this study, the first GH17 glucanosyltransglycosylase, MlGH17B, from a marine bacterium (Muricauda lutaonensis), was characterized. The enzyme was moderately thermostable with Tm at 64.4 °C and 73.2 °C, but an activity optimum at 20 °C, indicating temperature sensitive active site interactions. MlGH17B uses β-1,3 laminari-oligosaccharides with a degree of polymerization (DP) of 4 or higher as donors. Two glucose moieties (bound in the aglycone +1 and + 2 subsites) are cleaved off from the reducing end of the... (More)

The marine environment, contains plentiful renewable resources, e.g. macroalgae with unique polysaccharides, motivating search for enzymes from marine microorganisms to explore conversion possibilities of the polysaccharides. In this study, the first GH17 glucanosyltransglycosylase, MlGH17B, from a marine bacterium (Muricauda lutaonensis), was characterized. The enzyme was moderately thermostable with Tm at 64.4 °C and 73.2 °C, but an activity optimum at 20 °C, indicating temperature sensitive active site interactions. MlGH17B uses β-1,3 laminari-oligosaccharides with a degree of polymerization (DP) of 4 or higher as donors. Two glucose moieties (bound in the aglycone +1 and + 2 subsites) are cleaved off from the reducing end of the donor while the remaining part (bound in the glycone subsites) is transferred to an incoming β-1,3 glucan acceptor, making a β-1,6-linkage, thereby synthesizing branched or kinked oligosaccharides. Synthesized oligosaccharides up to DP26 were detected by mass spectrometry analysis, showing that repeated transfer reactions occurred, resulting in several β-1,6-linked branches. The modelled structure revealed an active site comprising five subsites: three glycone (-3, -2 and - 1) and two aglycone (+1 and + 2) subsites, with significant conservation of substrate interactions compared to the only crystallized 1,3-β-glucanosyltransferase from GH17 (RmBgt17A from the compost thriving fungus Rhizomucor miehei), suggesting a common catalytic mechanism, despite different phylogenetic origin, growth environment, and natural substrate. Both enzymes lacked the subdomain extending the aglycone subsites, found in GH17 endo-β-glucanases from plants, but this extension was also missing in bacterial endoglucanases (modelled here), showing that this feature does not distinguish transglycosylation from hydrolysis, but may rather relate to phylogeny.

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publication status
epub
subject
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Glycobiology
publisher
Oxford University Press
external identifiers
  • scopus:85190100528
  • pmid:38271624
ISSN
1460-2423
DOI
10.1093/glycob/cwae007
language
English
LU publication?
yes
id
5612f95f-22b8-4fd9-ac52-6d0d3ad8a030
date added to LUP
2024-02-01 05:10:11
date last changed
2024-04-23 10:58:17
@article{5612f95f-22b8-4fd9-ac52-6d0d3ad8a030,
  abstract     = {{<p>The marine environment, contains plentiful renewable resources, e.g. macroalgae with unique polysaccharides, motivating search for enzymes from marine microorganisms to explore conversion possibilities of the polysaccharides. In this study, the first GH17 glucanosyltransglycosylase, MlGH17B, from a marine bacterium (Muricauda lutaonensis), was characterized. The enzyme was moderately thermostable with Tm at 64.4 °C and 73.2 °C, but an activity optimum at 20 °C, indicating temperature sensitive active site interactions. MlGH17B uses β-1,3 laminari-oligosaccharides with a degree of polymerization (DP) of 4 or higher as donors. Two glucose moieties (bound in the aglycone +1 and + 2 subsites) are cleaved off from the reducing end of the donor while the remaining part (bound in the glycone subsites) is transferred to an incoming β-1,3 glucan acceptor, making a β-1,6-linkage, thereby synthesizing branched or kinked oligosaccharides. Synthesized oligosaccharides up to DP26 were detected by mass spectrometry analysis, showing that repeated transfer reactions occurred, resulting in several β-1,6-linked branches. The modelled structure revealed an active site comprising five subsites: three glycone (-3, -2 and - 1) and two aglycone (+1 and + 2) subsites, with significant conservation of substrate interactions compared to the only crystallized 1,3-β-glucanosyltransferase from GH17 (RmBgt17A from the compost thriving fungus Rhizomucor miehei), suggesting a common catalytic mechanism, despite different phylogenetic origin, growth environment, and natural substrate. Both enzymes lacked the subdomain extending the aglycone subsites, found in GH17 endo-β-glucanases from plants, but this extension was also missing in bacterial endoglucanases (modelled here), showing that this feature does not distinguish transglycosylation from hydrolysis, but may rather relate to phylogeny.</p>}},
  author       = {{Allahgholi, Leila and Derks, Maik G.N. and Dobruchowska, Justyna M and Jasilionis, Andrius and Moenaert, Antoine and Jouy, Léonie and Gulshan Kazi, Zubaida and Linares-Pastén, Javier A and Friðjónsson, Ólafur H and Hreggviðsson, Guðmundur Óli and Nordberg Karlsson, Eva}},
  issn         = {{1460-2423}},
  language     = {{eng}},
  month        = {{01}},
  publisher    = {{Oxford University Press}},
  series       = {{Glycobiology}},
  title        = {{Exploring a novel β-1,3-glucanosyltransglycosylase, <i>Ml</i>GH17B, from a marine <i>Muricauda lutaonensis</i> strain for modification of laminari-oligosaccharides}},
  url          = {{http://dx.doi.org/10.1093/glycob/cwae007}},
  doi          = {{10.1093/glycob/cwae007}},
  year         = {{2024}},
}