Metal-phenolic networks as tuneable spore coat mimetics

Bacillus subtilis are important probiotic microbes currently formulated for delivery as spores, but their ability to germinate in the gut remains debatable. To optimize their application, cells should be delivered in their vegetative state, but the sensitivity of B. subtilis prevents this. Through the application of self-assembled metal-phenolic network (MPN) cellular coatings, B. subtilis are protected from lyophilization stresses. These MPNs are an important class of self-assembled materials comprised of polyphenols and metal ions, and the efficacy of MPN protection was found to be dependent on the MPN components used for assembly. Both the size of the polyphenol and stability of the metal-phenol coordination were important factors that influenced their cellular protection; the smallest polyphenol, gallic acid, and the most stable chelated ion, Fe^III, were found to provide the highest level of protection. Further, delivery to the gut involves exposure to acidic conditions in the form of stomach acid and intestinal fluid. MPN coatings rapidly disassemble upon mild acid treatment but were found to protect B. subtilis from the negative impacts of the acid. Overall, optimized MPNs were found to protect vegetative B. subtilis cells from lyophilization stress and enable a more complete understanding of the role of each component in MPNs.