Works (1)
Updated: July 31st, 2023 21:14
2011 journal article
A high-throughput screen for directed evolution of aminocoumarin amide synthetases
Analytical Biochemistry, 419(1), 61–66.
author keywords: Directed evolution; Aminocoumarin; High-throughput screening; Natural products
MeSH headings : Acylation; Amide Synthases / metabolism; Aminocoumarins / chemistry; Aminocoumarins / metabolism; Chromatography, High Pressure Liquid; Cloning, Molecular; Directed Molecular Evolution / methods; Enzyme Activation; Escherichia coli / genetics; Escherichia coli / metabolism; Fluorescence; Fluorometry / methods; High-Throughput Screening Assays / methods; Molecular Structure; Novobiocin / analogs & derivatives; Novobiocin / chemistry; Novobiocin / metabolism; Plasmids / genetics; Plasmids / metabolism; Reproducibility of Results; Substrate Specificity
TL;DR:
A fluorimetric high-throughput screen is designed and validated, which can be used to report the activity of mutant amide synthetases toward a broad range of coumarin and acyl donor substrates, and is anticipated to accelerate the creation of amid synthetase mutants with new specificities by directed evolution.
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Added: August 6, 2018
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