Počet záznamů: 1  

Rutinosidase from Aspergillus niger: crystal structure and insight into the enzymatic activity

  1. 1.
    SYSNO ASEP0540276
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    Poddruh JČlánek ve WOS
    NázevRutinosidase from Aspergillus niger: crystal structure and insight into the enzymatic activity
    Tvůrce(i) Pachl, P. (CZ)
    Kapešová, J. (CZ)
    Brynda, Jiří (UMG-J) RID
    Biedermannova, L. (CZ)
    Pelantová, H. (CZ)
    Bojarová, P. (CZ)
    Křen, V. (CZ)
    Řezáčová, Pavlína (UMG-J) RID
    Kotik, M. (CZ)
    Celkový počet autorů9
    Zdroj.dok.FEBS Journal - ISSN 1742-464X
    Roč. 287, č. 15 (2020), s. 3315-3327
    Poč.str.13 s.
    Forma vydáníOnline - E
    Jazyk dok.eng - angličtina
    Země vyd.GB - Velká Británie
    Klíč. slovacatalytic mechanism ; diglycosidase ; rutin ; siras ; X-ray crystallography
    Vědní obor RIVEB - Genetika a molekulární biologie
    Obor OECDBiochemistry and molecular biology
    Způsob publikováníOmezený přístup
    Institucionální podporaUMG-J - RVO:68378050
    UT WOS000508932900001
    DOI10.1111/febs.15208
    AnotaceRutinosidases (alpha-l-rhamnosyl-beta-d-glucosidases) catalyze the cleavage of the glycosidic bond between the aglycone and the disaccharide rutinose (alpha-l-rhamnopyranosyl-(1> 6)-beta-d-glucopyranose) of specific flavonoid glycosides such as rutin (quercetin 3-O-rutinoside). Microbial rutinosidases are part of the rutin catabolic pathway, enabling the microorganism to utilize rutin and related plant phenolic glycosides. Here, we report the first three-dimensional structure of a rutinosidase determined at 1.27-angstrom resolution. The rutinosidase from Aspergillus niger K2 (AnRut), a member of glycoside hydrolase family GH-5, subfamily 23, was heterologously produced in Pichia pastoris. The X-ray structure of AnRut is represented by a distorted (beta/alpha)(8) barrel fold with its closest structural homologue being an exo-beta-(1,3)-glucanase from Candida albicans (CaExg). The catalytic site is located in a deep pocket with a striking structural similarity to CaExg. However, the entrance to the active site of AnRut has been found to be different from that of CaExg a mostly unstructured section of similar to 40 residues present in CaExg is missing in AnRut, whereas an additional loop of 13 amino acids partially covers the active site of AnRut. NMR analysis of reaction products provided clear evidence for a retaining reaction mechanism of AnRut. Unexpectedly, quercetin 3-O-glucoside was found to be a better substrate than rutin, and thus, AnRut cannot be considered a typical diglycosidase. Mutational analysis of conserved active site residues in combination with in silico modeling allowed identification of essential interactions for enzyme activity and helped to reveal further details of substrate binding. The protein sequence of AnRut has been revised.
    PracovištěÚstav molekulární genetiky
    KontaktNikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217
    Rok sběru2021
    Elektronická adresahttps://febs.onlinelibrary.wiley.com/doi/abs/10.1111/febs.15208
Počet záznamů: 1  

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