Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 103, Issue 6, Pages 1693-1698Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0504733102
Keywords
enzyme selection; reporter genes
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To determine whether microbial chemosensors can be used to find new or better biocatalysts, we constructed Escherichia coli hosts that recognize the product of a biocatalytic conversion through the transcriptional activator NahR and respond by expression of a lacZ or tetA reporter gene. Equipped with a benzaldehyde dehydrogenase (XyIC from Pseudomonas putida), the lacZ-based host responded to the oxidation of benzaldehyde and 2-hydroxybenzal-dehyde to the corresponding benzoic acids by forming blue colonies, whereas XyIC(-) cells did not. Similarly, the tetA-based host was able to grow under selective conditions only when equipped with XyIC, enabling selection of biocatalytically active cells in inactive populations at frequencies as low as 10(-6).
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