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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F19%3A00504360" target="_blank" >RIV/61388963:_____/19:00504360 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/19:00504360

Výsledek na webu

<a href="https://link.springer.com/article/10.1007%2Fs00253-018-9534-z" target="_blank" >https://link.springer.com/article/10.1007%2Fs00253-018-9534-z</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00253-018-9534-z" target="_blank" >10.1007/s00253-018-9534-z</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

—

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Applied Microbiology and Biotechnology

ISSN

0175-7598

e-ISSN

—

Svazek periodika

103

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

17

Strana od-do

1737-1753

Kód UT WoS článku

000459250200016

EID výsledku v databázi Scopus

2-s2.0-85059464051