Optimizing the immobilization conditions of beta-galactosidase on UV-cured epoxy-based polymeric film using response surface methodology


Beyler-Çigil A., Danis Ö. , Sarsar O., Kahraman M. V. , Ogan A. , Demir S.

JOURNAL OF FOOD BIOCHEMISTRY, vol.45, no.4, 2021 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 45 Issue: 4
  • Publication Date: 2021
  • Doi Number: 10.1111/jfbc.13699
  • Title of Journal : JOURNAL OF FOOD BIOCHEMISTRY
  • Keywords: &#946, &#8208, galactosidase, enzyme immobilization, epoxy&#8208, based polymeric film, response surface methodology

Abstract

UV-cured epoxy-based polymeric film was prepared from glycidyl methacrylate, trimethylolpropane triacrylate, and poly(ethylene glycol) methylether acrylate. 2-hydroxy-2- methylpropiophenone was used as photo initiator. Covalent binding through epoxy groups was employed to immobilize beta-galactosidase from Escherichia coli onto this film, and immobilization conditions were optimized by the response surface methodology. ATR-Fourier transform infrared (FTIR) and scanning electron microscopy (SEM) analysis was carried out to characterize the epoxy-based polymeric film. Immobilization yield of beta-galactosidase on the material was calculated as 3.57 mg/g and the highest enzyme activity for the immobilized enzyme recorded at pH 6.5 degrees C and 60 degrees C. The immobilized enzyme preserved 51% of its activity at the end of 12 runs. Free and immobilized enzyme hydrolyzed 163.8 and 172.3 mu M lactose from 1% lactose, respectively. Kinetic parameters of both free and immobilized beta-galactosidase were also investigated, and K-m values were determined to be 0.647 and 0.7263 mM, respectively.