Skip to Main content Skip to Navigation
Journal articles

Identification of the nucleophile catalytic residue of GH51 alpha-L-arabinofuranosidase from Pleurotus ostreatus

Abstract : In this study, the recombinant α-l-arabinofuranosidase from the fungus Pleurotus ostreatus (rPoAbf) was subjected to site-directed mutagenesis in order to identify the catalytic nucleophile residue. Based on bioinformatics and homology modelling analyses, E449 was revealed to be the potential nucleophilic residue. Thus, the mutant E449G of PoAbf was recombinantly expressed in Pichia pastoris and its recombinant expression level and reactivity were investigated in comparison to the wild-type. The design of a suitable set of hydrolysis experiments in the presence or absence of alcoholic arabinosyl acceptors and/or formate salts allowed to unambiguously identify the residue E449 as the nucleophile residue involved in the retaining mechanism of this GH51 arabinofuranosidase. 1H NMR analysis was applied for the identification of the products and the assignement of their anomeric configuration.
Complete list of metadata

Cited literature [29 references]  Display  Hide  Download

https://hal.archives-ouvertes.fr/hal-01439040
Contributor : Yves Bourne Connect in order to contact the contributor
Submitted on : Thursday, May 28, 2020 - 10:57:05 AM
Last modification on : Tuesday, October 19, 2021 - 10:35:21 PM

File

document.pdf
Publication funded by an institution

Licence


Distributed under a Creative Commons Attribution 4.0 International License

Identifiers

Collections

Citation

Antonella Amore, Alfonso Iadonisi, Florence Vincent, Vincenza Faraco. Identification of the nucleophile catalytic residue of GH51 alpha-L-arabinofuranosidase from Pleurotus ostreatus. AMB Express, Springer Nature, 2015, 5, ⟨10.1186/s13568-015-0164-x⟩. ⟨hal-01439040⟩

Share

Metrics

Record views

40

Files downloads

15