Acceptor Specificity of beta-N-Acetylhexosaminidase from Talaromyces flavus: A Rational Explanation
Identifikátory výsledku
Kód výsledku v IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68407700%3A21460%2F19%3A00336103" target="_blank" >RIV/68407700:21460/19:00336103 - isvavai.cz</a>
Výsledek na webu
<a href="https://doi.org/10.3390/ijms20246181" target="_blank" >https://doi.org/10.3390/ijms20246181</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.3390/ijms20246181" target="_blank" >10.3390/ijms20246181</a>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
Acceptor Specificity of beta-N-Acetylhexosaminidase from Talaromyces flavus: A Rational Explanation
Popis výsledku v původním jazyce
Fungal beta-N-acetylhexosaminidases, though hydrolytic enzymes in vivo, are useful tools in the preparation of oligosaccharides of biological interest. The beta-N-acetylhexosaminidase from Talaromyces flavus is remarkable in terms of its synthetic potential, broad substrate specificity, and tolerance to substrate modifications. It can be heterologously produced in Pichia pastoris in a high yield. The mutation of the Tyr470 residue to histidine greatly enhances its transglycosylation capability. The aim of this work was to identify the structural requirements of this model beta-N-acetylhexosaminidase for its transglycosylation acceptors and formulate a structure-activity relationship study. Enzymatic reactions were performed using an activated glycosyl donor, 4-nitrophenyl N-acetyl-beta-d-glucosaminide or 4-nitrophenyl N-acetyl-beta-d-galactosaminide, and a panel of glycosyl acceptors of varying structural features (N-acetylglucosamine, glucose, N-acetylgalactosamine, galactose, N-acetylmuramic acid, and glucuronic acid). The transglycosylation products were isolated and structurally characterized. The C-2 N-acetamido group in the acceptor molecule was found to be essential for recognition by the enzyme. The presence of the C-2 hydroxyl moiety strongly hindered the normal course of transglycosylation, yielding unique non-reducing disaccharides in a low yield. Moreover, whereas the gluco-configuration at C-4 steered the glycosylation into the beta(1-4) position, the galacto-acceptor afforded a beta(1-6) glycosidic linkage. The Y470H mutant enzyme was tested with acceptors based on beta-glycosides of uronic acid and N-acetylmuramic acid. With the latter acceptor, we were able to isolate and characterize one glycosylation product in a low yield.
Název v anglickém jazyce
Acceptor Specificity of beta-N-Acetylhexosaminidase from Talaromyces flavus: A Rational Explanation
Popis výsledku anglicky
Fungal beta-N-acetylhexosaminidases, though hydrolytic enzymes in vivo, are useful tools in the preparation of oligosaccharides of biological interest. The beta-N-acetylhexosaminidase from Talaromyces flavus is remarkable in terms of its synthetic potential, broad substrate specificity, and tolerance to substrate modifications. It can be heterologously produced in Pichia pastoris in a high yield. The mutation of the Tyr470 residue to histidine greatly enhances its transglycosylation capability. The aim of this work was to identify the structural requirements of this model beta-N-acetylhexosaminidase for its transglycosylation acceptors and formulate a structure-activity relationship study. Enzymatic reactions were performed using an activated glycosyl donor, 4-nitrophenyl N-acetyl-beta-d-glucosaminide or 4-nitrophenyl N-acetyl-beta-d-galactosaminide, and a panel of glycosyl acceptors of varying structural features (N-acetylglucosamine, glucose, N-acetylgalactosamine, galactose, N-acetylmuramic acid, and glucuronic acid). The transglycosylation products were isolated and structurally characterized. The C-2 N-acetamido group in the acceptor molecule was found to be essential for recognition by the enzyme. The presence of the C-2 hydroxyl moiety strongly hindered the normal course of transglycosylation, yielding unique non-reducing disaccharides in a low yield. Moreover, whereas the gluco-configuration at C-4 steered the glycosylation into the beta(1-4) position, the galacto-acceptor afforded a beta(1-6) glycosidic linkage. The Y470H mutant enzyme was tested with acceptors based on beta-glycosides of uronic acid and N-acetylmuramic acid. With the latter acceptor, we were able to isolate and characterize one glycosylation product in a low yield.
Klasifikace
Druh
J<sub>imp</sub> - Článek v periodiku v databázi Web of Science
CEP obor
—
OECD FORD obor
10401 - Organic chemistry
Návaznosti výsledku
Projekt
—
Návaznosti
V - Vyzkumna aktivita podporovana z jinych verejnych zdroju
Ostatní
Rok uplatnění
2019
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
International Journal of Molecular Sciences
ISSN
1422-0067
e-ISSN
1422-0067
Svazek periodika
20
Číslo periodika v rámci svazku
24
Stát vydavatele periodika
CH - Švýcarská konfederace
Počet stran výsledku
17
Strana od-do
—
Kód UT WoS článku
000506840100075
EID výsledku v databázi Scopus
2-s2.0-85076363105