Journal
PROCESS BIOCHEMISTRY
Volume 57, Issue -, Pages 95-104Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2017.03.025
Keywords
Human carbonic anhydrase; APTES; Magnetic nanoparticles; Immobilization; TEOS
Ask authors/readers for more resources
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. The immobilized enzyme activity was optimum at pH 8.0 and 25 degrees C. The Km value of the immobilized enzyme (1.02 mM) was higher than the free hCA I (0.48 mM). After 40 days incubation at 4 degrees C and 25 degrees C, the immobilized hCA I sustained 89% and 85% of its activity, respectively. Also, it sustained 61% of its initial activity after 13 cycles. Such results revealed good potential of immobilized enzyme for various applications.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available