Selective β-N-acetylhexosaminidase from Aspergillus versicolor-a tool for producing bioactive carbohydrates
Language English Country Germany Media print-electronic
Document type Journal Article
Grant support
LTC18038
Ministerstvo Školství, Mládeže a Tělovýchovy
LTC18041
Ministerstvo Školství, Mládeže a Tělovýchovy
LM2015042
CESNET Data Care
LM2015085
CERIT Scientific Cloud
PubMed
30603849
DOI
10.1007/s00253-018-9534-z
PII: 10.1007/s00253-018-9534-z
Knihovny.cz E-resources
- Keywords
- Aspergillus versicolor, Glycosidase, Heterologous expression, Homology modeling, Pichia pastoris, β-N-Acetylhexosaminidase,
- MeSH
- Aspergillus enzymology MeSH
- beta-N-Acetylhexosaminidases chemistry genetics isolation & purification metabolism MeSH
- Chromatography, Ion Exchange MeSH
- Disaccharides metabolism MeSH
- Gene Expression MeSH
- Catalytic Domain MeSH
- Protein Conformation MeSH
- Models, Molecular MeSH
- Pichia genetics metabolism MeSH
- Recombinant Proteins chemistry genetics isolation & purification metabolism MeSH
- Talaromyces enzymology MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- beta-N-Acetylhexosaminidases MeSH
- Disaccharides MeSH
- Recombinant Proteins MeSH
β-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 β-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 β-N-acetylhexosaminidases from other sources. The enzyme was used in a sequential reaction together with a mutant β-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-3 - a 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.
References provided by Crossref.org
The β-N-Acetylhexosaminidase in the Synthesis of Bioactive Glycans: Protein and Reaction Engineering
