4.4 Article

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

Journal

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 174, Issue 5, Pages 1834-1849

Publisher

HUMANA PRESS INC
DOI: 10.1007/s12010-014-1174-x

Keywords

Polyelectrolyte complex microcapsules; LentiKats (R); Confocal laser scanning microscopy; Inverse size exclusion chromatography; Environmental scanning electron microscopy; Baeyer-Villiger biooxidation

Funding

  1. Slovak Grant Agency for Science VEGA [1/0229/12]
  2. Slovak Research and Development Agency [APVV-0302-10, APVV-0486-10]
  3. Grant Agency of the Czech Republic [GA14-22777S]

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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 LentiKatsA (R) 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 LentiKatsA (R) 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 aEuro parts per thousand 2.75 nm for regular polyelectrolyte complex microcapsules and aEuro parts per thousand 1.89 nm for LentiKatsA (R) 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 x 10(-10) m(2) s(-1) for polyelectrolyte complex microcapsules and 0.36 x 10(-10) m(2) s(-1) for LentiKatsA (R). Recombinant cells Escherichia coli-overexpressing enzyme cyclopentanone monooxygenase were immobilised in polyelectrolyte complex microcapsules and LentiKatsA (R) 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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