Iron-Catalysed Radical Polymerisation by Living Bacteria.

The ability to harness cellular redox processes for abiotic synthesis might allow the preparation of engineered hybrid living systems. Towards this goal we describe a new bacteria-mediated iron-catalysed reversible deactivation radical polymerisation (RDRP), with a range of metal-chelating agents and monomers that can be used under ambient conditions with a bacterial redox initiation step to generate polymers. Cupriavidus metallidurans, Escherichia coli, and Clostridium sporogenes species were chosen for their redox enzyme systems and evaluated for their ability to induce polymer formation. Parameters including cell and catalyst concentration, initiator species, and monomer type were investigated. Water-soluble synthetic polymers were produced in the presence of the bacteria with full preservation of cell viability. This method provides a means by which bacterial redox systems can be exploited to generate "unnatural" polymers in the presence of "host" cells, thus setting up the possibility of making natural-synthetic hybrid structures and conjugates.

Oxygen-Tolerant RAFT Polymerization Initiated by Living Bacteria.

Living organisms can synthesize a wide range of macromolecules from a small set of natural building blocks, yet there is potential for even greater materials diversity by exploiting biochemical processes to convert unnatural feedstocks into new abiotic polymers. Ultimately, the synthesis of these polymers in situ might aid the coupling of organisms with synthetic matrices, and the generation of biohybrids or engineered living materials. The key step in biohybrid materials preparation is to harness the relevant biological pathways to produce synthetic polymers with predictable molar masses and defined architectures under ambient conditions. Accordingly, we report an aqueous, oxygen-tolerant RAFT polymerization platform based on a modified Fenton reaction, which is initiated by Cupriavidus metallidurans CH34, a bacterial species with iron-reducing capabilities. We show the synthesis of a range of water-soluble polymers under normoxic conditions, with control over the molar mass distribution, and also the production of block copolymer nanoparticles via polymerization-induced self-assembly. Finally, we highlight the benefits of using a bacterial initiation system by recycling the cells for multiple polymerizations. Overall, our method represents a highly versatile approach to producing well-defined polymeric materials within a hybrid natural-synthetic polymerization platform and in engineered living materials with properties beyond those of biotic macromolecules.

Engineering bacteria to control electron transport altering the synthesis of non-native polymer.

The use of bacteria as catalysts for radical polymerisations of synthetic monomers has recently been established. However, the role of trans Plasma Membrane Electron Transport (tPMET) in modulating these processes is not well understood. We sort to study this by genetic engineering a part of the tPMET system NapC in E. coli. We show that this engineering altered the rate of extracellular electron transfer coincided with an effect on cell-mediated polymerisation using a model monomer. A plasmid with arabinose inducible PBAD promoters were shown to upregulate NapC protein upon induction at total arabinose concentrations of 0.0018% and 0.18%. These clones (E. coli (IP_0.0018%) and E. coli (IP_0.18%), respectively) were used in iron-mediated atom transfer radical polymerisation (Fe ATRP), affecting the nature of the polymerisation, than cultures containing suppressed or empty plasmids (E. coli (IP_S) and E. coli (E), respectively). These results lead to the hypothesis that EET (Extracellular Electron Transfer) in part modulates cell instructed polymerisations.

Dual Stimuli-Responsive Comb Polymers from Modular N-Acylated Poly(aminoester)-Based Macromonomers.

We report the synthesis of dual-responsive N-acylated poly(aminoester) (NPAE)-based comb polymers with varying molecular composition and monomer sequence via a combination of spontaneous zwitterionic copolymerization and redox-initiated reversible addition-fragmentation chain transfer (RRAFT) polymerization. NPAE macromonomers were synthesized from different nucleophilic (MN), for example, 2-ethyl-2-oxazoline (EtOx) or 2-ethyl-2-oxazine (EtOz), and electrophilic monomers (ME), for example, acrylic acid (AA) or 2-carboxyethyl acrylate (CEA), to tune the hydrophilicity and sequence of the systems. The latter was found to influence the thermal properties and stability of the respective comb polymers. Turbidity investigations in aqueous solution revealed a dual-responsive behavior of the comb polymers being responsive to both temperature and pH changes due to ω-carboxylic end groups of the NPAE-based macromonomers. Additional methylene groups in the NPAE backbone rendered the corresponding systems more hydrophobic and, hence, decreased the cloud point temperatures and, at the same time, increased the pH values (at constant temperature) at which the polymer phase separates from the aqueous solution.