Immobilized bacteria are being extensively used for metabolite production, biocatalysts, and biosensor construction. However, long-term viability and metabolic activity of entrapped bacteria is affected by several conditions such as their physiological state, the presence of high-osmolarity environments, porous structure and shrinkage of the matrix. The aim of this work was to evaluate the effect of various parameters on bacteria immobilized in sol–gel-derived silica matrices. With this purpose, we evaluated the stress of immobilization over bacteria cultures obtained from different growing states, the effect of cell density and bacteria capability to proliferate inside matrices. Best results to attain longer preservation times were obtained when we immobilized suspensions with an optimized bacterial number of 1 × 107 cfu/gel in the presence of LB medium using aqueous silica precursors. Furthermore, the impact of osmotic stress with the subsequent intracellular trehalose accumulation and the addition of osmolites were investigated. Shorter preservation times were found for bacteria immobilized in the presence of osmolites while trehalose accumulation in stressed cells did not produce changes on entrapped bacteria viability. Finally, nutrient addition in silica matrices was studied indicating that the presence of a carbon source without the simultaneous addition of nitrogen was detrimental for immobilized E. coli. However, when both carbon and nitrogen sources were present, bacteria were able to survive longer periods of time.
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G.S.A. is grateful for her doctoral fellowship granted by the National Research Council (CONICET). M.L.F is grateful for her undergraduate fellowship granted by the University of Buenos Aires. The authors would like to acknowledge the support of grants from the Universidad of Buenos Aires UBACYT B049 (L.E.D.) and B407 (M.F.D.), and Agencia Nacional de Investigaciones Científicas y Técnicas BID 1728/OC-AR PICT 14192 (L.E.D.) and 32310 (M.F.D.).
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Alvarez, G.S., Foglia, M.L., Copello, G.J. et al. Effect of various parameters on viability and growth of bacteria immobilized in sol–gel-derived silica matrices. Appl Microbiol Biotechnol 82, 639–646 (2009). https://doi.org/10.1007/s00253-008-1783-9
- Escherichia coli
- Living cells