Improved thermal stability of phytase from Yersinia intermedia by physical adsorption immobilization on amino-multiwalled carbon nanotubes

• Published in: Bioprocess and biosystems engineering 2021-10, Vol.44 (10), p.2217-2228
• Main Authors: Lahiji, Sima, Hemmati, Roohullah, Homaei, Ahmad, Saffar, Behnaz, Ghorbani, Mansoureh
• Format: Article
• Language: English
• Subjects: 6-Phytase - isolation & purification; 6-Phytase - metabolism; Adsorption; Biotechnology; Chemistry; Chemistry and Materials Science; Deactivation; Enthalpy; Entropy; Environmental Engineering/Biotechnology; Enzyme Stability; Food Science; Free energy; Immobilization; Inactivation; Industrial and Production Engineering; Industrial Chemistry/Chemical Engineering; Kinetics; Microscopy, Electron, Scanning; Multi wall carbon nanotubes; Nanomaterials; Nanotechnology; Nanotubes; Nanotubes, Carbon - chemistry; Parameters; pH effects; Phosphorus; Phytase; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Research Paper; Scanning electron microscopy; Shelf life; Spectroscopy, Fourier Transform Infrared; Storage stability; Temperature; Thermal stability; Thermodynamics; Yersinia - enzymology; Yersinia intermedia; Zeta potential
• ISSN: 1615-7591; 1615-7605
• DOI: 10.1007/s00449-021-02598-4

Phytase is used in poultry diets to hydrolyze and release of phytate-bound phosphorus. Immobilization on nanomaterials optimizes enzyme’s thermal stability and reusability. This study aimed to immobilize the recombinant phytase from Yersinia intermedia on the surface of amino-multi-walled carbon nanotubes (amino-MWCNTs) by physical adsorption. For this, zeta potential measurement, FTIR spectroscopic analysis, scanning electron microscope (SEM), kinetic as well as thermodynamic parameters were used to characterize immobilized phytase on amino-MWCNTs. According to results, the optimum temperature of the immobilized phytase increased from 50 to 70 °C and also thermal and pH stability improved considerably. Moreover, immobilization led to an increase in the value of K m and k cat from 0.13 to 0.33 mM and 2220 to 2776 s −1 , respectively. In addition, the changes in activation energy of thermal inactivation (ΔE # a (D) ), the free energy of thermal inactivation (ΔG # D ) and the enthalpy of thermal inactivation (ΔH # D ) for immobilized phytase increased by +11.05, +24.7 and +11.4 kj/mole, respectively, while the value of the change in the entropy of thermal inactivation (ΔS # D ) decreased by − 0.04 kj/mole.K. Overall, our results showed that adsorption immobilization of phytase on amino-MWCNTs increases thermal, pH and storage stability as well as some of kinetic parameters.