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Physical and Bioengineering Properties of Polyvinyl Alcohol Lens-Shaped Particles Versus Spherical Polyelectrolyte Complex Microcapsules as Immobilisation Matrices for a Whole-Cell Baeyer–Villiger Monooxygenase

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Abstract

Direct comparison of key physical and chemical-engineering properties of two representative matrices for multipurpose immobilisations was performed for the first time. Polyvinyl alcohol lens-shaped particles LentiKats® and polyelectrolyte complex microcapsules were characterised by advanced techniques with respect to the size distribution of the particles, their inner morphology as revealed by fluorescent probe staining, mechanical resistance, size-exclusion properties, determination of effective diffusion coefficient and environmental scanning electron microscope imaging. While spherical polyelectrolyte complex microcapsules composed of a rigid semipermeable membrane and a liquid core are almost uniform in shape and size (diameter of 0.82 mm; RSD = 5.6 %), lens-shaped LentiKats® are characterised by wider size distribution (diameter of 3.65 mm; RSD = 10.3 % and height of 0.341 mm; RSD = 32.3 %) and showed the same porous structure throughout their whole volume at the mesoscopic (micrometre) level. Despite differences in their inner structure and surface properties, the pore diameter of ∼ 2.75 nm for regular polyelectrolyte complex microcapsules and ∼ 1.89 nm for LentiKats® were similar. These results were used for mathematical modelling, which provided the estimates of the effective diffusion coefficient of sucrose. This value was 1.67 × 10−10 m2 s−1 for polyelectrolyte complex microcapsules and 0.36 × 10−10 m2 s−1 for LentiKats®. Recombinant cells Escherichia coli-overexpressing enzyme cyclopentanone monooxygenase were immobilised in polyelectrolyte complex microcapsules and LentiKats® for comparison of their operational stability using model Baeyer–Villiger oxidation of (±)-cis-bicyclo [3.2.0] hept-2-en-6-one to regioisomeric lactones as important chiral synthons for potential pharmaceuticals. Both immobilisation matrices rendered high operational stability for whole-cell biocatalyst with no reduction in the biooxidation rate over 18 repeated reaction cycles.

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Acknowledgements

This work was supported by the Slovak Grant Agency for Science VEGA 1/0229/12, by the Slovak Research and Development Agency under contract No. APVV-0302-10 and No. APVV-0486-10 as well as Grant Agency of the Czech Republic GA14-22777S. We thank Prof. Marko D. Mihovilovic (VUT, Vienna, Austria) for providing the expression strain E. coli-overexpressing CPMO and Dr. Radek Stloukal from LentiKats (Czech Republic) for personal communication.

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Authors and Affiliations

  1. Institute of Chemistry, Centre for Glycomics, Slovak Academy of Sciences, Dúbravská cesta 9, 845 38, Bratislava, Slovakia

    Andrea Schenkmayerová, Marek Bučko & Peter Gemeiner

  2. Polymer Institute, Slovak Academy of Sciences, Dúbravská cesta 9, 845 41, Bratislava, Slovakia

    Dušana Treľová & Igor Lacík

  3. International Laser Centre, Ilkovičova 3, 841 04, Bratislava 4, Slovakia

    Dušan Chorvát Jr.

  4. Institute of Chemical and Environmental Engineering, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37, Bratislava, Slovakia

    Pavel Ačai & Milan Polakovič

  5. Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37, Bratislava, Slovakia

    Lukáš Lipták, Martin Rebroš & Michal Rosenberg

  6. Axxence Slovakia s.r.o., Mickiewiczova 9, 811 07, Bratislava, Slovakia

    Vladimír Štefuca

  7. ASCR, Institute of Scientific Instruments, Královopolska 147, 612 64, Brno, Czech Republic

    Vilém Neděla & Eva Tihlaříková

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  1. Andrea Schenkmayerová
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Correspondence to Andrea Schenkmayerová.

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Schenkmayerová, A., Bučko, M., Gemeiner, P. et al. Physical and Bioengineering Properties of Polyvinyl Alcohol Lens-Shaped Particles Versus Spherical Polyelectrolyte Complex Microcapsules as Immobilisation Matrices for a Whole-Cell Baeyer–Villiger Monooxygenase. Appl Biochem Biotechnol 174, 1834–1849 (2014). https://doi.org/10.1007/s12010-014-1174-x

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