Simone Krings

Biofilm bioreactors are attracting growing interest in the wastewater industry, as they allow higher cell densities and thus higher reaction rates compared to conventional bioreactors. However, some commonly used nitrifying bacteria, such as Nitrosomonas europaea, are slow-growing and need a prolonged period of time to develop a mature biofilm. Here, a biocoating or "living paint" is introduced, which is a synthetic biofilm made from a colloidal polymer (synthetic latex) binder encapsulating viable nitrifying bacteria at high density. Conventionally, the film formation of biocoatings is achieved by drying a bacteria/latex mixture. However, this fabrication is detrimental to the viability of the encapsulated bacteria because of the osmotic stress induced by desiccation. A nondesiccating film formation process is presented for biocoatings, which exploits two colloid science phenomena: coagulation and wet sintering. Desiccation-sensitive, nitrifying bacteria are employed in the biocoatings to convert NH4+ to NO2- and then NO3-. These biocoatings have a conversion rate (NO2- and NO3- production) of 3 mg N g(-1) d(-1) that is five times higher than in conventionally desiccated biocoatings. The reactivity continues over a period of 1 month. The processing method for these living paints is transformative for wastewater treatment and other applications using delicate, desiccation-sensitive microorganisms.

A biocoating confines non-growing, metabolically-active bacteria within a synthetic colloidal polymer (i.e. latex) film. Bacteria encapsulated inside biocoatings can perform useful functions, such as a biocatalyst in wastewater treatment. A biocoating needs to have high a permeability to allow a high rate of mass transfer for rehydration and the transport of both nutrients and metabolic products. It therefore requires an interconnected porous structure. Tuning the porosity architecture is a challenge. Here, we exploited rigid tubular nanoclays (halloysite) and non-toxic latex particles (with a relatively high glass transition temperature) as the colloidal “building blocks” to tailor the porosity inside biocoatings containing Escherichia coli bacteria as a model organism. Electron microscope images revealed inefficient packing of the rigid nanotubes and proved the existence of nanovoids along the halloysite/polymer interfaces. Single-cell observations using confocal laser scanning microscopy provided evidence for metabolic activity of the E. coli within the biocoatings through the expression of yellow fluorescent protein. A custom-built apparatus was used to measure the permeability of a fluorescein sodium salt in the biocoatings. Whereas there was no measurable permeability in a coating made from only latex particles, the permeability coefficient of the composite biocoatings increased with increasing halloysite content up to a value of 110-4 m h-1. The effects of this increase in permeability was demonstrated through a specially-developed resazurin reduction assay. Bacteria encapsulated in halloysite composite biocoatings had statistically significant higher metabolic activities in comparison to bacteria encapsulated in a non-optimized coating made from latex particles alone.