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1.
Biotechnol Prog ; 32(1): 43-53, 2016.
Article in English | MEDLINE | ID: mdl-26439442

ABSTRACT

The use of penicillin G acylase (PGA) covalently linked to insoluble carrier is expected to produce major advances in pharmaceutical processing industry and the enzyme stability enhancement is still a significant challenge. The objective of this study was to improve catalytic performance of the covalently immobilized PGA on a potential industrial carrier, macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [poly(GMA-co-EGDMA)], by optimizing the copolymerization process and the enzyme attachment procedure. This synthetic copolymer could be a very promising alternative for the development of low-cost, easy-to-prepare, and stable biocatalyst compared to expensive commercially available epoxy carriers such as Eupergit or Sepabeads. The PGA immobilized on poly(GMA-co-EGDMA) in the shape of microbeads obtained by suspension copolymerization appeared to have higher activity yield compared to copolymerization in a cast. Optimal conditions for the immobilization of PGA on poly(GMA-co-EGDMA) microbeads were 1 mg/mL of PGA in 0.75 mol/L phosphate buffer pH 6.0 at 25°C for 24 h, leading to the active biocatalyst with the specific activity of 252.7 U/g dry beads. Chemical amination of the immobilized PGA could contribute to the enhanced stability of the biocatalyst by inducing secondary interactions between the enzyme and the carrier, ensuring multipoint attachment. The best balance between the activity yield (51.5%), enzyme loading (25.6 mg/g), and stability (stabilization factor 22.2) was achieved for the partially modified PGA.


Subject(s)
Enzymes, Immobilized/chemistry , Enzymes , Penicillin Amidase/chemistry , Drug Industry , Enzyme Stability , Ethylene Glycols , Hydrogen-Ion Concentration , Methacrylates/chemistry , Microspheres , Polymers/chemistry
2.
Nanoscale Res Lett ; 5(1): 81-88, 2009 Oct 13.
Article in English | MEDLINE | ID: mdl-20652142

ABSTRACT

Surface modification of nanocrystalline TiO(2) particles (45 A) with bidentate benzene derivatives (catechol, pyrogallol, and gallic acid) was found to alter optical properties of nanoparticles. The formation of the inner-sphere charge-transfer complexes results in a red shift of the semiconductor absorption compared to unmodified nanocrystallites. The binding structures were investigated by using FTIR spectroscopy. The investigated ligands have the optimal geometry for chelating surface Ti atoms, resulting in ring coordination complexes (catecholate type of binuclear bidentate binding-bridging) thus restoring in six-coordinated octahedral geometry of surface Ti atoms. From the Benesi-Hildebrand plot, the stability constants at pH 2 of the order 10(3) M(-1) have been determined.

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