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

Result code in 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>

Alternative codes found

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

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

—

Czech description

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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

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2020

Confidentiality

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

Name of the periodical

FEBS Journal

ISSN

1742-464X

e-ISSN

—

Volume of the periodical

287

Issue of the periodical within the volume

15

Country of publishing house

GB - UNITED KINGDOM

Number of pages

13

Pages from-to

3315-3327

UT code for WoS article

000508932900001

EID of the result in the Scopus database

2-s2.0-85078675672