Lahiji Sima, Hemmati Roohullah, Homaei Ahmad, Saffar Behnaz, Ghorbani Mansoureh
Department of Biology, Faculty of Basic Sciences, Shahrekord University, Shahrekord, Iran.
Biotechnology Research Institute, Shahrekord University, Shahrekord, Iran.
Bioprocess Biosyst Eng. 2021 Oct;44(10):2217-2228. doi: 10.1007/s00449-021-02598-4. Epub 2021 Jun 17.
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 and k from 0.13 to 0.33 mM and 2220 to 2776 s, respectively. In addition, the changes in activation energy of thermal inactivation (ΔE), the free energy of thermal inactivation (ΔG) and the enthalpy of thermal inactivation (ΔH) 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) 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.
植酸酶用于家禽日粮中,以水解并释放与植酸盐结合的磷。固定在纳米材料上可优化酶的热稳定性和可重复使用性。本研究旨在通过物理吸附将中间耶尔森菌的重组植酸酶固定在氨基多壁碳纳米管(amino-MWCNTs)表面。为此,使用zeta电位测量、傅里叶变换红外光谱分析、扫描电子显微镜(SEM)、动力学以及热力学参数来表征固定在amino-MWCNTs上的植酸酶。根据结果,固定化植酸酶的最佳温度从50℃提高到70℃,热稳定性和pH稳定性也显著提高。此外,固定化导致K值和k值分别从0.13 mM增加到0.33 mM,从2220 s增加到2776 s。此外,固定化植酸酶热失活的活化能变化(ΔE)、热失活自由能(ΔG)和热失活焓(ΔH)分别增加了+11.05、+24.7和+11.4 kj/mol,而热失活熵变(ΔS)值降低了-0.04 kj/mol·K。总体而言,我们的结果表明,植酸酶在amino-MWCNTs上的吸附固定化提高了热稳定性、pH稳定性和储存稳定性以及一些动力学参数。