Abstract
Intracellular radical polymerizations allow for the direct bioorthogonal synthesis of various synthetic polymers within living cells, thereby providing a pathway to polymer-modified cells or the fermentative production of polymers. Here, we show that Escherichia coli cells can initiate the polymerization of various acrylamide, acrylic, and methacrylic monomers through an atom transfer radical reaction triggered by the activity of naturally occurring biomolecules within the bacterial cells. Intracellular radical polymerizations were confirmed by nuclear magnetic resonance spectroscopy, gel permeation chromatography of polymers extracted from the cells, and fluorescence labeling of the polymer directly inside the cells. The effect of polymerization on cell behavior and the response of the cells to polymerization was investigated through fluorescence microscopy and flow cytometry techniques, as well as metabolic and membrane integrity assays. The polymer synthesis and resulting products are cell-compatible, as indicated by the high viability of the polymerized cells. In cellulo synthesis of synthetic polymers containing fluorescent dyes was also achieved. These results not only enhance our understanding of the untapped potential of bacterial cells as living catalysts for polymer production but also reveal intracellular polymerization based on atom transfer radical polymerization initiators as a bioorthogonal tool for cell engineering and synthetic biology.