The beta-N-Acetylhexosaminidase in the Synthesis of Bioactive Glycans: Protein and Reaction Engineering

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F19%3A00503890" target="_blank" >RIV/61388971:_____/19:00503890 - isvavai.cz</a>

Result on the web

<a href="https://www.mdpi.com/1420-3049/24/3/599" target="_blank" >https://www.mdpi.com/1420-3049/24/3/599</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/molecules24030599" target="_blank" >10.3390/molecules24030599</a>

Result language

angličtina

Original language name

The beta-N-Acetylhexosaminidase in the Synthesis of Bioactive Glycans: Protein and Reaction Engineering

Original language description

N-Acetylhexosamine oligosaccharides terminated with GalNAc act as selective ligands of galectin-3, a biomedically important human lectin. Their synthesis can be accomplished by beta-N-acetylhexosaminidases (EC 3.2.1.52). Advantageously, these enzymes tolerate the presence of functional groups in the substrate molecule, such as the thiourea linker useful for covalent conjugation of glycans to a multivalent carrier, affording glyconjugates. beta-N-Acetylhexosaminidases exhibit activity towards both N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc) moieties. A point mutation of active-site amino acid Tyr into other amino acid residues, especially Phe, His, and Asn, has previously been shown to strongly suppress the hydrolytic activity of beta-N-acetylhexosaminidases, creating enzymatic synthetic engines. In the present work, we demonstrate that Tyr470 is an important mutation hotspot for altering the ratio of GlcNAcase/GalNAcase activity, resulting in mutant enzymes with varying affinity to GlcNAc/GalNAc substrates. The enzyme selectivity may additionally be manipulated by altering the reaction medium upon changing pH or adding selected organic co-solvents. As a result, we are able to fine-tune the beta-N-acetylhexosaminidase affinity and selectivity, resulting in a high-yield production of the functionalized GalNAc beta 4GlcNAc disaccharide, a selective ligand of galectin-3.

Czech name

—

Czech description

—

Type

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

CEP classification

—

OECD FORD branch

10608 - Biochemistry and molecular biology

Project

<a href="/en/project/LTC18038" target="_blank" >LTC18038: Multivalent glyco-biomaterials with a potential in regeneration medicine</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2019

Confidentiality

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

Name of the periodical

Molecules

ISSN

1420-3049

e-ISSN

—

Volume of the periodical

24

Issue of the periodical within the volume

3

Country of publishing house

CH - SWITZERLAND

Number of pages

14

Pages from-to

599

UT code for WoS article

000458934000223

EID of the result in the Scopus database

2-s2.0-85061365096