Selection of biocatalysts for chemical synthesis
2006; National Academy of Sciences; Volume: 103; Issue: 6 Linguagem: Inglês
10.1073/pnas.0504733102
ISSN1091-6490
AutoresStephan van Sint Fiet, Jan B. van Beilen, Bernard Witholt,
Tópico(s)Enzyme Catalysis and Immobilization
ResumoTo 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 (XylC from Pseudomonas putida), the lacZ-based host responded to the oxidation of benzaldehyde and 2-hydroxybenzaldehyde to the corresponding benzoic acids by forming blue colonies, whereas XylC- cells did not. Similarly, the tetA-based host was able to grow under selective conditions only when equipped with XylC, enabling selection of biocatalytically active cells in inactive populations at frequencies as low as 10(-6).
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