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Cryostructured and Crosslinked Viable Cells Forming Monoliths Suitable for Bioreactor Applications

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Abstract

Applications of immobilized biocatalysts in both research and industry require highly active catalysts, preferably at a low cost. In this study, cryogels with high catalyst density were produced through cryostructuration of whole Escherichia coli (E.coli) cells. Prepared cryogels are macroporous materials composed of metabolically active cells crosslinked to each other via polymeric structures. Different macromolecular reagents: oxidized dextran (OxDex), polyvinyl alcohol (PVA) and polyethyleneimine (PEI), the two latter activated with glutaraldehyde (GTA) have been synthesized. Prepared polymers were tested as effective and mild crosslinkers for cells during the cryostructuration procedure. Combination of the two synthetic polymers: PEI+GTA and PVA+GTA was found most suitable for formation of macroporous and stable structures from cells without any toxic effect on them. About 90 % of β-glucosidase activity in cells was retained after crosslinking with a combination of synthetic polymers, whereas E. coli crosslinked using GTA showed complete loss of activity. Preserved viability of cells in cryogel offers possibility to induce protein expression in cells after crosslinking. For β-glucosidase induction post immobilization yielded 50 % activity of that from cells induced in free form before cryo-structurization. The results of the post immobilization studies indicate an interesting potential for handling very sensitive enzymes.

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Acknowledgments

To funding of Marie Curie Networks for Initial Training fellowship in the BIOTRAINS project (FP7-PEOPLE-ITN-2008-238531) and the financial support from Ångpanneföreningens Foundation for Research and Development is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, 221 00, Lund, Sweden

    Oksana Zaushitsyna, Dmitriy Berillo, Harald Kirsebom & Bo Mattiasson

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  1. Oksana Zaushitsyna
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  2. Dmitriy Berillo
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  3. Harald Kirsebom
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  4. Bo Mattiasson
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Correspondence to Bo Mattiasson.

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Zaushitsyna, O., Berillo, D., Kirsebom, H. et al. Cryostructured and Crosslinked Viable Cells Forming Monoliths Suitable for Bioreactor Applications. Top Catal 57, 339–348 (2014). https://doi.org/10.1007/s11244-013-0189-9

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  • DOI: https://doi.org/10.1007/s11244-013-0189-9

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