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 Escherichia coli strain DH10B increment pyrFEC and the use of N-4-benzoyl-2'-deoxycytidine as a sole sou...

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Veröffentlicht in:Catalysts 2020-04, Vol.10 (4), p.445, Article 445
Hauptverfasser: Urbeliene, Nina, Meskiene, Rita, Tiskus, Matas, Stanislauskiene, Ruta, Aucynaite, Agota, Laurynenas, Audrius, Meskys, Rolandas
Format: Artikel
Sprache:eng
Schlagworte:
amidase
amidohydrolase
Amino acids
Biocatalysts
Catalysts
Chemical reactions
Chemistry
Chemistry, Physical
Cloning
Deoxyribonucleic acid
DNA
E coli
Enzymes
Gene expression
Genetic testing
Genomes
high-throughput screening
Homology
metagenomic libraries
Methods
Microorganisms
Physical Sciences
Plasmids
Polyethylene terephthalate
Proteins
Science & Technology
selection method
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Zusammenfassung: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 Escherichia coli strain DH10B increment pyrFEC and the use of N-4-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
2073-4344
DOI:10.3390/catal10040445