School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, India.
Int J Biol Macromol. 2011 Nov 1;49(4):674-80. doi: 10.1016/j.ijbiomac.2011.06.027. Epub 2011 Jul 5.
Ureases isolated from leguminous sources, Canavalia ensiformis and Cajanus cajan were immobilized onto gold nanoparticles (nano-ureases). Optimization of the urease immobilization was carried using response surface methodology based on Central Composite Design. Immobilization efficiency of nano-urease from C. ensiformis and C. cajan were found to be 215.10% and 255.92%, respectively. The methodology adopted has deviation of 2.56% and 3.01% with respect to experimental values in case of C. ensiformis and C. cajan, respectively. Nano-urease from C. cajan has broad physico-chemical parameters with pH optimum from 7.1 to 7.3 and temperature optimum from 50 to 70°C. Nano-urease from C. ensiformis has sharp pH and temperature optima at 7.3 and 70°C, respectively. Fourier transform infra-red spectroscopy has revealed involvement of groups viz. amino, glycosyl moiety, etc. in urease immobilization onto gold nano-particles. Transmission and scanning electron micrographs revealed that arrangement of urease onto gold nano-particles from C. ensiformis was uniform while it was localized in case of C. cajan. Nano-urease from C. ensiformis has higher specificity and catalysis toward urea as compared to nano-urease from C. cajan. Nano-ureases from both sources are equally stable for 6 months under dried conditions and can be used for 10 washes.
从豆科植物剑豆(Canavalia ensiformis)和木豆(Cajanus cajan)中分离出的脲酶被固定在金纳米粒子(纳米脲酶)上。基于中心复合设计的响应面法对脲酶固定化进行了优化。发现来自 C. ensiformis 和 C. cajan 的纳米脲酶的固定化效率分别为 215.10%和 255.92%。采用的方法在 C. ensiformis 和 C. cajan 的情况下,相对于实验值有 2.56%和 3.01%的偏差。来自 C. cajan 的纳米脲酶具有较宽的物理化学参数,最佳 pH 值为 7.1 至 7.3,最佳温度为 50 至 70°C。来自 C. ensiformis 的纳米脲酶的 pH 和温度最佳值分别为 7.3 和 70°C。傅里叶变换红外光谱显示,氨基、糖基等基团参与了脲酶在金纳米粒子上的固定化。透射和扫描电子显微镜显示,来自 C. ensiformis 的脲酶在金纳米粒子上的排列是均匀的,而来自 C. cajan 的脲酶则是局部的。与来自 C. cajan 的纳米脲酶相比,来自 C. ensiformis 的纳米脲酶对尿素具有更高的特异性和催化活性。来自这两种来源的纳米脲酶在干燥条件下稳定 6 个月,可以使用 10 次洗涤。