Počet záznamů: 1
The beta-N-Acetylhexosaminidase in the Synthesis of Bioactive Glycans: Protein and Reaction Engineering
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SYSNO ASEP 0503890 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název The beta-N-Acetylhexosaminidase in the Synthesis of Bioactive Glycans: Protein and Reaction Engineering Tvůrce(i) Bojarová, Pavla (MBU-M) ORCID
Kulik, Natalia (MBU-M) ORCID
Hovorková, Michaela (MBU-M)
Slámová, Kristýna (MBU-M) RID, ORCID
Pelantová, Helena (MBU-M) ORCID, RID
Křen, Vladimír (MBU-M) RID, ORCIDČíslo článku 599 Zdroj.dok. Molecules. - : MDPI
Roč. 24, č. 3 (2019)Poč.str. 14 s. Jazyk dok. eng - angličtina Země vyd. CH - Švýcarsko Klíč. slova beta-N-acetylhexosaminidase ; galectin-3 ; site-directed mutagenesis Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP LTC18038 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Způsob publikování Open access Institucionální podpora MBU-M - RVO:61388971 UT WOS 000458934000223 EID SCOPUS 85061365096 DOI 10.3390/molecules24030599 Anotace 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. Pracoviště Mikrobiologický ústav Kontakt Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Rok sběru 2020 Elektronická adresa https://www.mdpi.com/1420-3049/24/3/599
Počet záznamů: 1