Journal
ACS SYNTHETIC BIOLOGY
Volume 1, Issue 11, Pages 526-531Publisher
AMER CHEMICAL SOC
DOI: 10.1021/sb300010a
Keywords
enzyme; photocontrol; cell-free expression; beta-lactamase; AzoTAB
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Funding
- European Research Council (ERC) [European Community] [258782]
- ENS de Lyon
- European Research Council (ERC) [258782] Funding Source: European Research Council (ERC)
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beta-lactams can be converted into beta-amino acids by beta-lactamase, a bacterial enzyme, leading to significant change in the biological function of the substrate molecules. Here we describe a method for photocontrol of beta-lactam conversion without gene nor enzyme modification. This is achieved by the addition of,a Cationic photosensitive surfactant, AzoTAB, to a gene expression Medium containing DNA Coding for beta-lactamase, the enzyme capable of the desired conversion. In the absence of UV (365 nm) or after illumination by blue light (480 nm) for 4-min, conversion of beta-lactam is strongly reduced. while the application of UV for 4 mm results in a strong enhancement of Substrate conversion. Several cycles of activation! 1 inhibition are obtained upon successive UV/blue light illuminations. When both reconstituted photoresponsive gene expression. medium and beta-lactamase substrate are encapsulated in independent microfluidic chambers, selective UV illumination results in spatially resolved activation of substrate conversion.
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