Circular dichroism analysis of penicillin G acylase covalently immobilized on silica nanoparticles

Kranz, Bertolt; Bürck, Jochen; Franzreb, Matthias; Köster, Rainer; Ulrich, Anne S.

doi:10.1016/j.jcis.2007.08.062

Cited by 23 publications

(23 citation statements)

References 34 publications

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“…Therefore, the initial concentration of PGA of 1 mg/mL in attachment solution was set as optimal and used for further studies. The results are similar to the results of previous studies concerning immobilization of PGA on silica nanoparticles, where the specific activity determined by a kinetic assay decreased by around 60%, when increasing the enzyme loading from 14.9 to 55.8 mg/g . Because of the fact that no major changes in the secondary structure have been found, diffusion limitations appeared to be the main reason for the decline of the activity …”

Section: Resultssupporting

confidence: 90%

An approach for the improved immobilization of penicillin G acylase onto macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) as a potential industrial biocatalyst

Knežević‐Jugović

Žuža

Jakovetić

et al. 2015

Biotechnol Progress

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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.

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Section: Resultssupporting

confidence: 90%

An approach for the improved immobilization of penicillin G acylase onto macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) as a potential industrial biocatalyst

Knežević‐Jugović

Žuža

Jakovetić

et al. 2015

Biotechnol Progress

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“…Silica, as a support for nano-immobilization has several advantages of biocompatibility, chemical inertness and ease of functionalization [7]. Important enzymes like penicillin G acylase [30] and b-1,4-glucosidase [27] have been successfully immobilized on silica nanoparticles.…”

Section: Resultsmentioning

confidence: 99%

Immobilization of halophilic Bacillus sp. EMB9 protease on functionalized silica nanoparticles and application in whey protein hydrolysis

Sinha

Khare

2014

Bioprocess Biosyst Eng

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The present work targets the fabrication of an active, stable, reusable enzyme preparation using functionalized silica nanoparticles as an effective enzyme support for crude halophilic Bacillus sp. EMB9 protease. The immobilization efficiency under optimized conditions was 60%. Characterization of the immobilized preparation revealed marked increase in pH and thermal stability. It retained 80% of its original activity at 70 °C while t 1/2 at 50 °C showed a five-fold enhancement over that for the free protease. Kinetic constants K m and V max were indicative of a higher reaction velocity along with decreased affinity for substrate. The preparation could be efficiently reused up to 6 times and successfully hydrolysed whey proteins with high degree of hydrolysis. Immobilization of a crude halophilic protease on a nanobased scaffold makes the process cost effective and simple.

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“…Amine-modified nanoparticles can be prepared using different organosilane precursor reagents for cohydrolysis or post-coating [137][138][139][140][141][142][143][144]. Biofunctionalization with amine-containing ligands requires an amine-reactive homobifunctional cross-linker (such as glutaraldehyde) and subsequent reduction of the formed Schiff base (using, for example, sodium cyanoborohydride).…”

Section: Covalent Bindingmentioning

confidence: 99%

Review: Bioanalytical applications of biomolecule-functionalized nanometer-sized doped silica particles

Knopp

Tang

Nießner

2009

Analytica Chimica Acta

365

213

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Circular dichroism analysis of penicillin G acylase covalently immobilized on silica nanoparticles

Cited by 23 publications

References 34 publications

An approach for the improved immobilization of penicillin G acylase onto macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) as a potential industrial biocatalyst

An approach for the improved immobilization of penicillin G acylase onto macroporous poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) as a potential industrial biocatalyst

Immobilization of halophilic Bacillus sp. EMB9 protease on functionalized silica nanoparticles and application in whey protein hydrolysis

Review: Bioanalytical applications of biomolecule-functionalized nanometer-sized doped silica particles

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