Department of Chemistry, Faculty of Science and Arts, Inonu University, 44280, Malatya, Turkey.
Appl Biochem Biotechnol. 2019 Mar;187(3):938-956. doi: 10.1007/s12010-018-2853-9. Epub 2018 Aug 13.
This paper describes a new support that permits to efficient immobilization of L-asparaginase (L-ASNase). For this purpose, FeO magnetic nanoparticles were synthesized and coated by MCM-41. 3-chloropropyltrimethoxysilane (CPTMS) was used as a surface modifying agent for covalent immobilization of L-ASNase on the magnetic nanoparticles. The chemical structure; thermal, morphological, and magnetic properties; chemical composition; and zeta potential value of FeO@MCM-41-Cl were characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), differential thermal analysis (DTA), differential scanning calorimetry (DSC), vibrating sample magnetometer (VSM), scanning electron microscope (SEM), energy dispersive X-ray (EDX), X-ray diffraction patterns (XRD), and zeta-potential measurement. The immobilization efficiency onto FeO@MCM-41-Cl was detected as 63%. The reusability, storage, pH, and thermal stabilities of the immobilized L-ASNase were investigated and compared to that of soluble one. The immobilized enzyme maintained 42.2% of its original activity after 18 cycles of reuse. Furthermore, it was more stable towards pH and temperature compared with soluble enzyme. The Michaelis-Menten kinetic properties of immobilized L-ASNase showed a lower V and a similar K compared to soluble L-ASNase. Immobilized enzyme had around 47 and 32.5% residual activity upon storage a period of 28 days at 4 and 25 °C, respectively. In conclusion, the FeO@MCM-41-Cl@L-ASNase core-shell nanoparticles could successfully be used in industrial and medical applications.
本文描述了一种新的载体,可有效固定 L-天冬酰胺酶(L-ASNase)。为此,合成了 FeO 磁性纳米粒子,并将其包覆在 MCM-41 上。3-氯丙基三甲氧基硅烷(CPTMS)用作将 L-ASNase共价固定在磁性纳米粒子上的表面改性剂。通过傅里叶变换红外光谱(FTIR)、热重分析(TGA)、差热分析(DTA)、差示扫描量热法(DSC)、振动样品磁强计(VSM)、扫描电子显微镜(SEM)、能量色散 X 射线(EDX)、X 射线衍射图谱(XRD)和 ζ 电位测量对 FeO@MCM-41-Cl 的化学结构、热、形态和磁性特性、化学组成和 ζ 电位值进行了表征。固定化效率检测为 63%。对 FeO@MCM-41-Cl 固定化 L-ASNase 的可重复使用性、储存稳定性、pH 值和热稳定性进行了研究,并与可溶性 L-ASNase 进行了比较。固定化酶在 18 次重复使用后仍保持其原始活性的 42.2%。此外,它在 pH 值和温度方面比可溶性酶更稳定。固定化 L-ASNase 的米氏动力学特性表明,V 值较低,K 值与可溶性 L-ASNase 相似。固定化酶在 4°C 和 25°C 下储存 28 天时,剩余活性分别约为 47%和 32.5%。总之,FeO@MCM-41-Cl@L-ASNase 核壳纳米粒子可成功应用于工业和医疗领域。