title: An approach for the improved immobilization of penicillin G acylase onto macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) as a potential industrial biocatalyst.
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.
authors.affiliation: Dept. of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Zorica D
authors.initials: ZD
authors.lastname: Knežević-Jugović
authors.affiliation: Dept. of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Milena G
authors.initials: MG
authors.lastname: Žuža
authors.affiliation: Dept. of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Sonja M
authors.initials: SM
authors.lastname: Jakovetić
authors.affiliation: Dept. of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Andrea B
authors.initials: AB
authors.lastname: Stefanović
authors.affiliation: Dept. of Physical Chemistry and Electrochemistry, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Enis S
authors.initials: ES
authors.lastname: Džunuzović
authors.affiliation: Dept. of Physical Chemistry and Electrochemistry, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Katarina B
authors.initials: KB
authors.lastname: Jeremić
authors.affiliation: Dept. of Physical Chemistry and Electrochemistry, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Serbia.
authors.forename: Slobodan M
authors.initials: SM
authors.lastname: Jovanović
mesh_terms: D004345:Drug Industry; D004795:Enzyme Stability; D004798:Enzymes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005026:Ethylene Glycols; D006863:Hydrogen-Ion Concentration; D008689:Methacrylates / Q000737:chemistry; D008863:Microspheres; D010399:Penicillin Amidase / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry
chemical_list: D004798:Enzymes; D004800:Enzymes, Immobilized; D005026:Ethylene Glycols; D008689:Methacrylates; D011108:Polymers; C526534:poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate); C065843:Eupergit C; D010399:Penicillin Amidase
keywords: Eupergit C; chemical amination; immobilization; penicillin G acylase; poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate); suspension copolymerization