A Rapid Method for the Selection of Amidohydrolases from Metagenomic Libraries by Applying Synthetic Nucleosides and a Uridine Auxotrophic Host

A Rapid Method for the Selection of Amidohydrolases from Metagenomic Libraries by Applying Synthetic Nucleosides and a Uridine Auxotrophic Host

In this study, the development of a rapid, high-throughput method for the selection of amide-hydrolysing enzymes from the metagenome is described. This method is based on uridine auxotrophic <i>Escherichia coli</i> strain DH10B ∆<i>pyrFEC</i> and the use of <i>N</i&g...

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Bibliographic Details
Main Authors: Nina Urbelienė, Rita Meškienė, Matas Tiškus, Rūta Stanislauskienė, Agota Aučynaitė, Audrius Laurynėnas, Rolandas Meškys
Format: Article
Language:English
Published: MDPI AG 2020-04-01
Series:Catalysts
Subjects:
metagenomic libraries
amidohydrolase
amidase
selection method
high-throughput screening
Online Access:https://www.mdpi.com/2073-4344/10/4/445
Description
Summary:In this study, the development of a rapid, high-throughput method for the selection of amide-hydrolysing enzymes from the metagenome is described. This method is based on uridine auxotrophic <i>Escherichia coli</i> strain DH10B ∆<i>pyrFEC</i> and the use of <i>N</i><sup>4</sup>-benzoyl-2’-deoxycytidine as a sole source of uridine in the minimal microbial M9 medium. The approach described here permits the selection of unique biocatalysts, e.g., a novel amidohydrolase from the activating signal cointegrator homology (ASCH) family and a polyethylene terephthalate hydrolase (PETase)-related enzyme.
ISSN:2073-4344

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