Abstract
A colourimetric high-throughput screening system was established for directed evolution of prodigiosin ligase PigC. The two-step system consists of a colony prescreening test and a subsequent photometric 96-well plate assay. Screening PigC epPCR libraries in Pseudomonas putida revealed a PigC variant that achieved a 2.9× increased yield of prodiginine derivatives.
MeSH terms
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Colorimetry
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Directed Molecular Evolution*
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Escherichia coli / metabolism
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High-Throughput Screening Assays / methods*
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Kinetics
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Ligases / genetics
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Ligases / metabolism*
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Mutagenesis, Site-Directed
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Prodigiosin / metabolism
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Pseudomonas putida / metabolism
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Substrate Specificity
Substances
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Bacterial Proteins
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Ligases
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Prodigiosin