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
2014
Schenkmayerová, Andrea | Bučko, Marek | Gemeiner, Peter | Treľová, Dušana | Lacík, Igor | Chorvát, Dušan, Jr | Ačai, Pavel | Polakovič, Milan | Lipták, Lukáš | Rebroš, Martin | Rosenberg, Michal | Štefuca, Vladimír | Neděla, Vilém | Tihlaříková, Eva
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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