@article{article_557021, title={Optimization of the Immobilization Conditions of Horseradish Peroxidase on TiO2-COOH nanoparticles by Box-Behnken Design}, journal={Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi}, volume={23}, pages={904–916}, year={2019}, DOI={10.19113/sdufenbed.557021}, url={https://izlik.org/JA99EN35AW}, author={Şahin, Selmihan}, keywords={İmmobilizasyon,Horseradish peroksidaz,TiO2 nanopartiküller,EDC/NHS bağlama,Karboksil grubu fonksiyonelleştirmesi}, abstract={<span lang="en-us" style="font-size:11pt;line-height:115%;font-family:Cambria, serif;" xml:lang="en-us">In this
study, TiO <sub>2 </sub> nanoparticles were prepared and -COOH functionalized
with 3-(3,4-dihydroxyphenyl) propionic acid. The characterization of
nanoparticles was performed by FTIR, TEM, EDS and XRD. HRP was immobilized on
those nanoparticles by EDC/NHS coupling reaction. The immobilization conditions
of HRP including A: enzyme concentration (0.5-1.5 mg/mL), B: immobilization pH
(4.0-8.0), C: immobilization temperature (4-50°C), D: immobilization time (1-20
h) were optimized by response surface methodology and Box-Behnken design. The
optimized immobilization conditions were identified as 0.5 mg/mL HRP, at pH
5.5, 40 °C for 8 h for activity of immobilized HRP, 1.5 mg/mL HRP, at pH 4 and
18°C for 20 h for protein binding yield (%). At these optimum conditions, the experimental
value for the activity of immobilized HRP was 80.39 U ± 1.06; protein binding
yield was 94.25 ± 3.58%. Moreover, the optimum temperature and pH of free and
immobilized enzyme were determined as 50°C and 4.0; 50°C and 3.5, respectively.
The activity of the immobilized HRP sustained 52% of its initial activity after
10 days storage at 4°C </span> <span style="font-size:11pt;line-height:115%;font-family:Cambria, serif;">. </span> <span lang="en-us" style="font-size:11pt;line-height:115%;font-family:Cambria, serif;" xml:lang="en-us">Furthermore,
the immobilized HRP sustained 48% of its initial activity after 6 consecutive
reactions.  </span> <br />}, number={3}