Description
Birgitte Zeuner, Marlène Vuillemin, Jan Muschiol, Eduardo S. Moreno Prieto, Kristian Barrett, Jesper Holck, Jens Preben Morth, Anne S. Meyer
Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads 221, DK-2800 Kgs. Lyngby, Denmark.
E-mail: [email protected]
Finding lacto-N-biosidases (LNBases; EC 3.2.1.140) in CAZy family GH20 appears to be like finding a needle in a haystack. Of the >13,000 sequences listed in GH20, 129 are characterized and 28 have resolved 3D structures (August 2022). The vast majority of these are confirmed β-N-acetylhexosaminidases (EC 3.2.1.52). Only two entries are con-firmed LNBases, and only one of these has a resolved structure. The LNBase ability to remove or transfer disaccharide moieties in a single reaction step is an attractive trait in glycoengineering. Here we present work on discovery of novel LNBases in GH20 using a combination of sequence analysis and structural information.In addition, we recently engineered the well-characterized LNBase from Bifidobacterium bifidum (LnbB) to increase its transglycosylation performance. Certain mutations to im-prove transglycosylation are likely transferable to the newly discovered GH20 LNBases. Using a combination of chemical synthesis, NMR analysis, specific enzymatic digestion and LC-ESI-MS, we were able to establish the regioselectivity exhibited by the LnbB var-iants in the transglycosylation to form lacto-N-tetraose (LNT) [1].
[1]Vuillemin M.; Holck J.; Matwiejuk M.; Moreno Prieto E.S.; Muschiol J.; Molnar-Gabor D.; Meyer A.S.; Zeuner B. Applied Sciences 2021, 11, 11493.
(Oral presentation)
| Period | 26 Sept 2022 |
|---|---|
| Event title | 14th Carbohydrate Bioengineering Meeting |
| Event type | Conference |
| Conference number | 14 |
| Location | Norefjell, NorwayShow on map |
| Degree of Recognition | International |