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.

Design and synthesis study of the thermo‐sensitive copolymer carrier of penicillin G acylase

“…Based on the rising importance of carrier on immobilized PGA, the design and preparation of carrier have been reported extensively . According to those reports, PGA immobilization carriers can be divided into 2 broad categories, the inorganic materials and organic materials.…”

The engineering and immobilization of penicillin G acylase onto thermo‐sensitive tri‐block copolymer system

“…At present, the loading capacity ( L ), activity retention ratio ( Ar ), enzyme activity ( Ea ), response rate ( Rr ), and reusable stability ( Rs ) are main problems of immobilized PGA, because they directly affect the economic efficiency of enterprises in actual production . According to reports, the structure and composition of the carrier play an important role in above performances, so exploring and synthesizing carrier with the best suitable composition and structure have become a long‐standing goal. Although significant effort has been made on immobilized carrier for PGA, the balance among L , Ar , Rr , and Rs was still not resolved well.…”

Efficient synthesis of cefadroxil in [Bmim][NTf2]‐phosphate cosolvent by magnetic immobilized penicillin G acylase

“…The absorption peak located at 636 cm −1 is the characteristic absorption of Fe 3 O 4 , which showed the presence of Fe 3 O 4 particles in polymer microspheres. The absorption peak located at 904 cm −1 is the characteristic absorption of the epoxy groups . The results of chemical analysis indicated that PM@Fe 3 O 4 had epoxy groups of 0.11 mM/g.…”