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
| Autor: | Alžbeta Marček Chorvátová, Florian Rudroff, Marietta Hakarová, Tomáš Krajčovič, Marek Bučko, Peter Gemeiner, Igor Lacík, Dusan Chorvat, Marko D. Mihovilovic |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
biocatalysis Reductase lcsh:Chemical technology medicine.disease_cause 01 natural sciences Catalysis lcsh:Chemistry 03 medical and health sciences Cascade reaction medicine lcsh:TP1-1185 Physical and Theoretical Chemistry Escherichia coli polyelectrolyte Alcohol dehydrogenase chemistry.chemical_classification biology 010405 organic chemistry Chemistry whole-cell biocatalyst multimodal optical imaging Polyelectrolyte 0104 chemical sciences 030104 developmental biology Enzyme lcsh:QD1-999 Permeability (electromagnetism) Yield (chemistry) immobilization biology.protein Biophysics enzyme cascade reaction |
| Zdroj: | Catalysts Volume 10 Issue 11 Catalysts, Vol 10, Iss 1288, p 1288 (2020) |
| ISSN: | 2073-4344 |
| DOI: | 10.3390/catal10111288 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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