Počet záznamů: 1
Rutinosidase from Aspergillus niger: crystal structure and insight into the enzymatic activity
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SYSNO ASEP 0540276 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Rutinosidase 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 autorů 9 Zdroj.dok. FEBS Journal - ISSN 1742-464X
Roč. 287, č. 15 (2020), s. 3315-3327Poč.str. 13 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova catalytic mechanism ; diglycosidase ; rutin ; siras ; X-ray crystallography Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Biochemistry and molecular biology Způsob publikování Omezený přístup Institucionální podpora UMG-J - RVO:68378050 UT WOS 000508932900001 DOI 10.1111/febs.15208 Anotace Rutinosidases (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. Pracoviště Ústav molekulární genetiky Kontakt Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Rok sběru 2021 Elektronická adresa https://febs.onlinelibrary.wiley.com/doi/abs/10.1111/febs.15208
Počet záznamů: 1