One-step formation of three-dimensional macroporous bacterial sponges as a novel approach for the preparation of bioreactors for bioremediation and green treatment of water.

Immobilisation of bacteria on or into a polymer support is a common method for the utilisation of bacteria as biocatalysts for many biotechnological, medical and environmental applications. The main challenge in this approach is the time taken for the formation of stable biofilms, and the typically low percentage of bacterial cells present on or in the polymer matrix. In this work we propose a novel method for producing a porous bacteria based structure with the properties of a sponge (bacterial sponge) that we then use as a bioreactor for water treatment. Cryogelation has been used as a tool to create macroporous ( i.e. with pores in the range 10-100 μm), highly permeable systems with low diffusion constraints and high bacterial content (more than 98% to total material content). A novel crosslinking system was used to form stable bacterial sponges with a high percentage of live bacteria organized in a 3D porous structure. The bacterial sponge was produced in a one step process and can be made from one or several bacterial strains (in this case, two bacterial strains Pseudomonas mendocina and Rhodoccocus koreensis (and a mixture of both) were used). Reduction of the total polymer content to 2% makes the system more sustainable and environmentally friendly under disposal as it can be simply composted. The bacterial sponges have good mechanical stability and cell viability, which enables repeated use of the materials for phenol degradation for up to five weeks. The material can be stored and transported in cryogenic conditions (-80 °C) for prolonged periods of time, retaining its bioremediation activity following 4-6 weeks of frozen storage. The proposed method of producing bioreactors with a high number of live immobilised bacteria, low polymer content and controlled 3D structure is a promising tool for developing novel materials based on active bacterial cells for various environmental, biotechnological, biological and medical applications.