Selective Beta-N-acetylhexosaminidase from Aspergillus versicolor—a tool for producing bioactive carbohydrates

0504360 - MBÚ 2020 RIV US eng J - Článek v odborném periodiku
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
Grant CEP: GA MŠMT(CZ) LTC18038; GA MŠMT(CZ) LTC18041
Výzkumná infrastruktura: CESNET II - 90042; CERIT-SC - 90085
Institucionální podpora: RVO:61388971 ; RVO:61388963
Klíčová slova: Aspergillus versicolor * Beta-N-Acetylhexosaminidase * Glycosidase
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 3.530, rok: 2019
Způsob publikování: Omezený přístup
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.
Trvalý link: http://hdl.handle.net/11104/0296013