Development of silk fibroin-based beads for immobilized cell fermentations

Silk fibroin was evaluated as a new matrix for immobilized cell fermentation. Silk fibroin was extracted from Bombyx mori cocoon, purified, concentrated in polyethylene glycol solution and diluted to 3 wt% with distilled water. This fibroin solution was used to encapsulate sensitive cells of the probiotic strain, Bifidobacterium longum ATCC 15707. Polymer droplets produced with an encapsulator were collected in liquid nitrogen and lyophilized. A low overall survival of 0.2% was measured after lyophilization. Lyophilized beads were hardened for 24 h under vacuum with an atmosphere of 89% relative humidity. The inoculated beads were colonized in two successive batch fermentations. Structure of silk fibroin beads and colonization of cells were examined with scanning electron microscopy. Colonized beads were tested in continuous fermentations for cell production. A biomass productivity of 1.7 x 10(9) CFU ml(-1) h(-1) was achieved, which was limited by loss of bead structure. This instability might be due to bead degradation by proteolytic activity of cells and/or limited mechanical stability during continuous fermentation in the stirred tank reactor.