Immobilization and characterization of human carbonic anhydrase I on amine functionalized magnetic nanoparticles

Al-Dhrub A. H. A., ŞAHİN S., ÖZMEN İ., Tunca E., Bulbul M.

PROCESS BIOCHEMISTRY, vol.57, pp.95-104, 2017 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 57
  • Publication Date: 2017
  • Doi Number: 10.1016/j.procbio.2017.03.025
  • Journal Name: PROCESS BIOCHEMISTRY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.95-104
  • Süleyman Demirel University Affiliated: Yes

Abstract

In this study, we synthesized magnetic nanoparticles (MNPs) by co-precipitation method. After that, silica coating with tetraethyl orthosilicate (TEOS) (SMNPs), amine functionalization of silica coated MNPs (ASMNPs) by using 3-aminopropyltriethoxysilane (APTES) were performed, respectively. After activation with glutaraldehyde (GA) of ASMNPs, human carbonic anhydrase (hCA I) was immobilized on ASMNPs. The characterization of nanoparticles was performed by transmission electron microscopy (TEM), fourier transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRD) and vibrating sample magnetometer (VSM). The immobilization conditions such as GA concentration, activation time of support with GA, enzyme amount, enzyme immobilization time were optimized. In addition of that, optimum conditions for activity, kinetic parameters (K-m, V-max, k(cat), k(cat)/K-m), thermal stability, storage stability and reusability of immobilized enzyme were determined.