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Selective Beta-N-acetylhexosaminidase from Aspergillus versicolor—a tool for producing bioactive carbohydrates
- 1.0504360 - MBÚ 2020 RIV US eng J - Journal Article
Bojarová, Pavla - Kulik, Natalia - Slámová, Kristýna - Hubálek, Martin - Kotík, Michael - Cvačka, Josef - Pelantová, Helena - Křen, Vladimír
Selective Beta-N-acetylhexosaminidase from Aspergillus versicolor—a tool for producing bioactive carbohydrates.
Applied Microbiology and Biotechnology. Roč. 103, č. 4 (2019), s. 1737-1753. ISSN 0175-7598. E-ISSN 1432-0614
R&D Projects: GA MŠMT(CZ) LTC18038; GA MŠMT(CZ) LTC18041
Research Infrastructure: CESNET II - 90042; CERIT-SC - 90085
Institutional support: RVO:61388971 ; RVO:61388963
Keywords : Aspergillus versicolor * Beta-N-Acetylhexosaminidase * Glycosidase
OECD category: Biochemistry and molecular biology
Impact factor: 3.530, year: 2019
Method of publishing: Limited access
https://link.springer.com/article/10.1007%2Fs00253-018-9534-z
Beta-N-Acetylhexosaminidases (EC 3.2.1.52) are typical of their dual activity encompassing both N-acetylglucosamine and N-acetylgalactosamine substrates. Here we present the isolation and characterization of a selective -N-acetylhexosaminidase from the fungal strain of Aspergillus versicolor. The enzyme was recombinantly expressed in Pichia pastoris KM71H in a high yield and purified in a single step using anion-exchange chromatography. Homologous molecular modeling of this enzyme identified crucial differences in the enzyme active site that may be responsible for its high selectivity for N-acetylglucosamine substrates compared to fungal -N-acetylhexosaminidases from other sources. The enzyme was used in a sequential reaction together with a mutant -N-acetylhexosaminidase from Talaromyces flavus with an enhanced synthetic capability, affording a bioactive disaccharide bearing an azido functional group. The azido function enabled an elegant multivalent presentation of this disaccharide on an aromatic carrier. The resulting model glycoconjugate is applicable as a selective ligand of galectin-3a biomedically attractive human lectin. These results highlight the importance of a general availability of robust and well-defined carbohydrate-active enzymes with tailored catalytic properties for biotechnological and biomedical applications.
Permanent Link: http://hdl.handle.net/11104/0296013
Number of the records: 1