M. Bucko, P. Gemeiner, T. Krajcovic, M. Hakarova, D. Chorvat, A. Chorvatova, I. Lacik, F. Rudroff, M.D. Mihovilovic:
"Immobilized cell physiology imaging and stabilization of enzyme cascade reaction using recombinant cells escherichia coli entrapped in polyelectrolyte complex beads by jet break-up encapsulator";
A novel, high performance, and scalable immobilization protocol using a laminar jet break-up technique was developed for the production of polyelectrolyte complex beads with entrapped viable Escherichia coli cells expressing an enzyme cascade of alcohol dehydrogenase, enoate reductase, and cyclohexanone monooxygenase. A significant improvement of operational stability was achieved by cell immobilization, which was manifested as an almost two-fold higher summative product yield of 63% after five cascade reaction cycles as compared to the yield using free cells of 36% after the maximum achievable number of three cycles. Correspondingly, increased metabolic activity was observed by multimodal optical imaging in entrapped cells, which was in contrast to a complete suppression of cell metabolism in free cells after five reaction cycles. Additionally, a high density of cells entrapped in beads had a negligible effect on bead permeability for low molecular weight substrates and products of cascade reaction.
enzyme cascade reaction; immobilization; polyelectrolyte; multimodal optical imaging; biocatalysis; whole-cell biocatalyst
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