Immobilisation of α-amylase on activated amidrazone acrylic fabric: a new approach for the enhancement of enzyme stability and reusability

Sci Rep. 2019 Sep 3;9(1):12672. doi: 10.1038/s41598-019-49206-w.

Abstract

In this study, amidrazone acrylic fabric was applied as an immobilising support for α-amylase. The immobilised α-amylase was characterised by Fourier transform infrared spectroscopy and scanning electron microscopy. Furthermore, the optimum conditions for immobilisation efficiency, immobilisation time, reusability, kinetic parameters and pH, for the immobilisation process were examined. The study demonstrated that with 4% cyanuric chloride, and a pH of 7.0, the highest immobilization efficiency of 81% was obtained. Around 65% of the initial activity was maintained after storage at 4 °C for 8 weeks. The immobilised enzyme retained 53% of its original activity after being reused 15 times and exhibited improved stability compared with the free enzyme in relation to heavy metal ions, pH, temperature and inhibitors. The immobilised enzyme presented kinetic parameters of 2.6 mg starch and 0.65 µmol maltose/mL for Km and Vmax respectively, compared with 3.7 mg starch and 0.83 µmol maltose/ mL for the free enzyme. The improvements in the enzyme's catalytic properties, stability and reusability obtained from immobilisation make amidrazone acrylic fabric support a good promising candidate for bio-industrial applications.

MeSH terms

  • Bacillus subtilis / enzymology
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Biocatalysis
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / metabolism
  • Enzyme Stability
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / metabolism
  • Hydrogen-Ion Concentration
  • Kinetics
  • Maltose / metabolism
  • Metals, Heavy / chemistry
  • Starch / metabolism
  • Temperature
  • Triazines / metabolism
  • alpha-Amylases / chemistry
  • alpha-Amylases / metabolism*

Substances

  • Bacterial Proteins
  • Enzyme Inhibitors
  • Enzymes, Immobilized
  • Metals, Heavy
  • Triazines
  • cyanuric chloride
  • Maltose
  • Starch
  • alpha-Amylases