Diasi Manab, Singh Rinki, Mahapatra Amarjyoti Das, L Renuka, Patel Hitarth, Ganatra Hasit, Datta Bhaskar
Department of Chemistry, Indian Institute of Technology Gandhinagar Palaj Gandhinagar 382355 Gujarat India
Blasto Research Private Limited Ahmedabad Gujarat India.
RSC Adv. 2024 Feb 27;14(10):6972-6984. doi: 10.1039/d3ra07606g. eCollection 2024 Feb 21.
In this work, we have studied the ability of urease immobilized on glutaraldehyde crosslinked chitosan coated magnetic iron oxide nanoparticles (Urease/GA/CS/MIONPs), for the hitherto unreported comparative hydrolysis of urea in synthetic (SUr) and real human urine (HUr). The prepared Urease/GA/CS/MIONPs were characterized by a combination of Fourier transform infrared spectroscopy (FTIR), field emission-scanning-electron-microscopy (FESEM), energy dispersive X-ray spectroscopy (EDX) and dynamic light scattering (DLS). The nanoconstructs display the highest ammonium ion liberation post-urea hydrolysis in 1/20 or 1/24-fold dilutions of SUr and HUr, respectively. The optimum activity of immobilized urease is observed at pH 7, and the nanoconstructs facilitate efficient urea-hydrolysis till at least 45 °C. Kinetic analysis of the immobilized urease shows and of 14.81 mM, 12.36 mM, and 18.55 μM min and 10.10 μM min, towards SUr and HUr, respectively. The magnetization of the immobilized urease is suitable for reuse across multiple cycles of urea hydrolysis in SUr and HUr. The robust performance of Urease/GA/CS/MIONPs in SUr and HUr is promising for generating ammonium as a useable source of nitrogen from human urine, and underscores the suitability of SUr as a urine mimic for such interventions.
在本研究中,我们研究了固定在戊二醛交联壳聚糖包覆磁性氧化铁纳米颗粒(脲酶/戊二醛/壳聚糖/磁性氧化铁纳米颗粒)上的脲酶对合成尿素(SUr)和真实人类尿液(HUr)中尿素进行前所未有的比较水解的能力。通过傅里叶变换红外光谱(FTIR)、场发射扫描电子显微镜(FESEM)、能量色散X射线光谱(EDX)和动态光散射(DLS)相结合的方法对制备的脲酶/戊二醛/壳聚糖/磁性氧化铁纳米颗粒进行了表征。纳米结构在SUr和HUr的1/20或1/24倍稀释液中尿素水解后显示出最高的铵离子释放量。固定化脲酶在pH 7时观察到最佳活性,并且纳米结构促进高效的尿素水解直至至少45°C。固定化脲酶的动力学分析表明,对SUr和HUr的米氏常数(Km)分别为14.81 mM、12.36 mM,最大反应速率(Vmax)分别为18.55 μM min和10.10 μM min。固定化脲酶的磁化适合在SUr和HUr中进行多个尿素水解循环的重复使用脲酶/戊二醛/壳聚糖/磁性氧化铁纳米颗粒在SUr和HUr中的强大性能有望从人类尿液中产生铵作为可用的氮源,并强调了SUr作为此类干预的尿液模拟物的适用性。