Transglycosidase activity of glycosynthase-type mutants of a fungal GH20 beta-N-acetylhexosaminidase
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F20%3A00533177" target="_blank" >RIV/61388971:_____/20:00533177 - isvavai.cz</a>
Result on the web
<a href="https://www.sciencedirect.com/science/article/pii/S0141813020334747" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0141813020334747</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.ijbiomac.2020.05.273" target="_blank" >10.1016/j.ijbiomac.2020.05.273</a>
Alternative languages
Result language
angličtina
Original language name
Transglycosidase activity of glycosynthase-type mutants of a fungal GH20 beta-N-acetylhexosaminidase
Original language description
beta-N-Acetylhexosaminidases (CAZy GH20, EC 3.2.1.52) are exo-glycosidases specific for cleaving N-acetylglucosamine and N-acetylgalactosamine moieties of various substrates. The beta-N-acetylhexosaminidase from the filamentous fungus Talaromyces flavus (TfHex), a model enzyme in this study, has a broad substrate flexibility and outstanding synthetic ability. We have designed and characterized seven glycosynthase-type variants of TfHex mutated at the catalytic aspartate residue that stabilizes the oxazoline reaction intermediate. Most of the obtained enzyme variants lost the majority of their original hydrolytic activity towards the standard substrate p-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside (pNP-beta-GlcNAc), moreover, the mutants were not active with the proposed glycosynthase donor 2-acetamido-2-deoxy-D-glucopyranosyl-alpha-fluoride (GlcNAc-alpha-F) either as would be expected in a glycosynthase. Importantly, the mutant enzymes instead retained a strong transglycosylation activity towards the standard substrate pNP-beta-GlcNAc. In summary, five out of seven prepared TfHex variants bearing mutation at the catalytic Asp370 residue acted as efficient transglycosidases, which makes them excellent tools for the synthesis of chitooligosaccharides, with the advantage of processing an inexpensive, stable and commercially available pNP-beta-GlcNAc.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10606 - Microbiology
Result continuities
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
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
International Journal of Biological Macromolecules
ISSN
0141-8130
e-ISSN
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Volume of the periodical
161
Issue of the periodical within the volume
OCT15
Country of publishing house
GB - UNITED KINGDOM
Number of pages
10
Pages from-to
1206-1215
UT code for WoS article
000569753300005
EID of the result in the Scopus database
2-s2.0-85086941152