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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F20%3A00521889" target="_blank" >RIV/61388963:_____/20:00521889 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/86652036:_____/20:00521889 RIV/61388971:_____/20:00521889 RIV/68378050:_____/20:00540276

Výsledek na webu

<a href="https://febs.onlinelibrary.wiley.com/doi/full/10.1111/febs.15208" target="_blank" >https://febs.onlinelibrary.wiley.com/doi/full/10.1111/febs.15208</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1111/febs.15208" target="_blank" >10.1111/febs.15208</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

Rutinosidases (α‐l ‐rhamnosyl‐β‐d ‐glucosidases) catalyze the cleavage of the glycosidic bond between the aglycone and the disaccharide rutinose (α‐l ‐rhamnopyranosyl‐(1→6)‐β‐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‐Å resolution. The rutinosidase from Aspergillus niger K2 (An Rut), a member of glycoside hydrolase family GH‐5, subfamily 23, was heterologously produced in Pichia pastoris . The X‐ray structure of An Rut is represented by a distorted (β/α)8 barrel fold with its closest structural homologue being an exo‐β‐(1,3)‐glucanase from Candida albicans (Ca Exg). The catalytic site is located in a deep pocket with a striking structural similarity to Ca Exg. However, the entrance to the active site of An Rut has been found to be different from that of Ca Exg – a mostly unstructured section of ~ 40 residues present in Ca Exg is missing in An Rut, whereas an additional loop of 13 amino acids partially covers the active site of An Rut. NMR analysis of reaction products provided clear evidence for a retaining reaction mechanism of An Rut. Unexpectedly, quercetin 3‐O‐glucoside was found to be a better substrate than rutin, and thus, An Rut 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 An Rut has been revised.

Název v anglickém jazyce

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

Popis výsledku anglicky

Rutinosidases (α‐l ‐rhamnosyl‐β‐d ‐glucosidases) catalyze the cleavage of the glycosidic bond between the aglycone and the disaccharide rutinose (α‐l ‐rhamnopyranosyl‐(1→6)‐β‐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‐Å resolution. The rutinosidase from Aspergillus niger K2 (An Rut), a member of glycoside hydrolase family GH‐5, subfamily 23, was heterologously produced in Pichia pastoris . The X‐ray structure of An Rut is represented by a distorted (β/α)8 barrel fold with its closest structural homologue being an exo‐β‐(1,3)‐glucanase from Candida albicans (Ca Exg). The catalytic site is located in a deep pocket with a striking structural similarity to Ca Exg. However, the entrance to the active site of An Rut has been found to be different from that of Ca Exg – a mostly unstructured section of ~ 40 residues present in Ca Exg is missing in An Rut, whereas an additional loop of 13 amino acids partially covers the active site of An Rut. NMR analysis of reaction products provided clear evidence for a retaining reaction mechanism of An Rut. Unexpectedly, quercetin 3‐O‐glucoside was found to be a better substrate than rutin, and thus, An Rut 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 An Rut has been revised.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

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

Název periodika

FEBS Journal

ISSN

1742-464X

e-ISSN

—

Svazek periodika

287

Číslo periodika v rámci svazku

15

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

13

Strana od-do

3315-3327

Kód UT WoS článku

000508932900001

EID výsledku v databázi Scopus

2-s2.0-85078675672