Surface-functionalized hyperbranched poly(amido acid) magnetic nanocarriers for covalent immobilization of a bacterial γ-glutamyltranspeptidase

Molecules. 2014 Apr 22;19(4):4997-5012. doi: 10.3390/molecules19044997.

Abstract

In this study, we synthesized water-soluble hyperbranched poly(amido acid)s (HBPAAs) featuring multiple terminal CO2H units and internal tertiary amino and amido moieties and then used them in conjunction with an in situ Fe2+/Fe3+ co-precipitation process to prepare organic/magnetic nanocarriers comprising uniformly small magnetic iron oxide nanoparticles (NP) incorporated within the globular HBPAAs. Transmission electron microscopy revealed that the HBPAA-γ-Fe2O3 NPs had dimensions of 6-11 nm, significantly smaller than those of the pristine γ-Fe2O3 (20-30 nm). Subsequently, we covalently immobilized a bacterial γ-glutamyltranspeptidase (BlGGT) upon the HBPAA-γ-Fe2O3 nanocarriers through the formation of amide linkages in the presence of a coupling agent. Magnetization curves of the HBPAA-γ-Fe2O3/BlGGT composites measured at 300 K suggested superparamagnetic characteristics, with a saturation magnetization of 52 emu g⁻¹. The loading capacity of BlGGT on the HBPAA-γ-Fe2O3 nanocarriers was 16 mg g⁻¹ support; this sample provided a 48% recovery of the initial activity. The immobilized enzyme could be recycled 10 times with 32% retention of the initial activity; it had stability comparable with that of the free enzyme during a storage period of 63 days. The covalent immobilization and stability of the enzyme and the magnetization provided by the HBPAA-γ-Fe2O3 NPs suggests that this approach could be an economical means of depositing bioactive enzymes upon nanocarriers for BlGGT-mediated bio-catalysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Enzyme Stability
  • Enzymes, Immobilized / chemistry*
  • Equipment Reuse
  • Escherichia coli / chemistry
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Ferric Compounds / chemistry*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Magnetite Nanoparticles / chemistry*
  • Nylons / chemistry*
  • Particle Size
  • Recombinant Proteins / chemistry
  • Temperature
  • gamma-Glutamyltransferase / chemistry*

Substances

  • Bacterial Proteins
  • Enzymes, Immobilized
  • Ferric Compounds
  • Magnetite Nanoparticles
  • Nylons
  • Recombinant Proteins
  • ferric oxide
  • gamma-Glutamyltransferase