Application of the Uridine Auxotrophic Host and Synthetic Nucleosides for a Rapid Selection of Hydrolases from Metagenomic Libraries

Overview

Journal Microb Biotechnol

Specialties Biotechnology
Microbiology

Date 2018 Oct 11

PMID 30302933

Citations 4

Authors

Nina Urbeliene

Simonas Kutanovas

Rita Meskiene

Renata Gasparaviciute

Daiva Tauraite

Martyna Koplunaite

Rolandas Meskys

Affiliations

Soon will be listed here.

Abstract

A high-throughput method (≥ 10 of clones can be analysed on a single agar plate) for the selection of ester-hydrolysing enzymes was developed based on the uridine auxotrophy of Escherichia coli strain DH10B ΔpyrFEC and the acylated derivatives 2',3',5'-O-tri-acetyluridine and 2',3',5'-O-tri-hexanoyluridine as the sole source of uridine. The proposed approach permits the selection of hydrolases belonging to different families and active towards different substrates. Moreover, the ester group of the substrate used for the selection, at least partly, determined the specificity of the selected enzymes.

Citing Articles

α-L-Fucosidases from an Alpaca Faeces Metagenome: Characterisation of Hydrolytic and Transfucosylation Potential.

Krupinskaite A, Stanislauskiene R, Serapinas P, Rutkiene R, Gasparaviciute R, Meskys R Int J Mol Sci. 2024; 25(2).

PMID: 38255883 PMC: 10815079. DOI: 10.3390/ijms25020809.

Cytidine deaminases catalyze the conversion of (,)-substituted pyrimidine nucleosides.

Urbeliene N, Tiskus M, Tamulaitiene G, Gasparaviciute R, Lapinskaite R, Jauniskis V Sci Adv. 2023; 9(5):eade4361.

PMID: 36735785 PMC: 9897663. DOI: 10.1126/sciadv.ade4361.

Identification of a 2'--Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries.

Aucynaite A, Rutkiene R, Tauraite D, Meskys R, Urbonavicius J Molecules. 2018; 23(11).

PMID: 30405065 PMC: 6278475. DOI: 10.3390/molecules23112904.

Application of the uridine auxotrophic host and synthetic nucleosides for a rapid selection of hydrolases from metagenomic libraries.

Urbeliene N, Kutanovas S, Meskiene R, Gasparaviciute R, Tauraite D, Koplunaite M Microb Biotechnol. 2018; 12(1):148-160.

PMID: 30302933 PMC: 6302743. DOI: 10.1111/1751-7915.13316.

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