Hypertransglycosylating Variants of the GH20 beta-N-Acetylhexosaminidase for the Synthesis of Chitooligomers

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F22%3A00558832" target="_blank" >RIV/61388971:_____/22:00558832 - isvavai.cz</a>

Result on the web

<a href="https://onlinelibrary.wiley.com/doi/epdf/10.1002/adsc.202200046" target="_blank" >https://onlinelibrary.wiley.com/doi/epdf/10.1002/adsc.202200046</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/adsc.202200046" target="_blank" >10.1002/adsc.202200046</a>

Result language

angličtina

Original language name

Hypertransglycosylating Variants of the GH20 beta-N-Acetylhexosaminidase for the Synthesis of Chitooligomers

Original language description

Fungal beta-N-acetylhexosaminidases of the CAZy family 20 of glycoside hydrolases are well-established tools for the enzymatic synthesis of a wide variety of natural and modified oligosaccharides and glycoconjugates. In order to increase their synthetic efficiency, the beta-N-acetylhexosaminidase from Aspergillus oryzae (AoHex) was employed as a model enzyme for enzyme engineering aiming at shifting the reaction course from hydrolysis toward transglycosylation. Specifically, nine mutant variants of AoHex were designed by molecular modeling based on its crystal structure and molecular dynamics simulations. The selected mutation hotspots included the tyrosine residue at the active site, which stabilizes the transition state of the reaction, and two residues at the aglycone-binding site, which were replaced by tryptophan residues to increase the hydrophobicity of this subsite. Besides the individual mutants, combined double-mutant variants were also prepared and characterized. As a result, eight out of the studied new AoHex variants had transglycosidase activity, with V306W/Y445N AoHex being a superior transglycosidase with a transglycosylation-to-hydrolysis ratio greater than 110, which is entirely unique among the hypertransglycosylating glycosidase mutants including the GH20 beta-N-acetylhexosaminidases.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2022

Confidentiality

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

Name of the periodical

Advanced Synthesis & Catalysis

ISSN

1615-4150

e-ISSN

1615-4169

Volume of the periodical

364

Issue of the periodical within the volume

12

Country of publishing house

DE - GERMANY

Number of pages

14

Pages from-to

2009-2022

UT code for WoS article

000796769700001

EID of the result in the Scopus database

2-s2.0-85130509136