Modification-Free Photocontrol of beta-Lactam Conversion with Spatiotemporal Resolution

作者：Venancio Marques Anna; Liu Yan Jun; Diguet Antoine; di Maio Thomas; Gautier Arnaud; Baigl Damien^*

来源：ACS Synthetic Biology, 2012, 1(11): 526-531.

DOI：10.1021/sb300010a

• enzyme
• photocontrol
• cell-free expression
• beta-lactamase
• AzoTAB

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.

• 出版日期2012-11