Farzi-Khajeh Hamed, Safa Kazem D, Dastmalchi Siavoush
Organosilicon Research Laboratory, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
Organosilicon Research Laboratory, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Nov 15;1068-1069:210-217. doi: 10.1016/j.jchromb.2017.10.041. Epub 2017 Oct 20.
Currently, modified FeO magnetic nanoparticles frequently are used as nanocarriers for proteins and enzymes purification. In the present study, FeO magnetic nanoparticles were prepared, and their surfaces were modified with p-aminophenol affinity ligand immobilized by different linkers. The modified nanocarriers were used for the purification of α-amylase from the bovine milk (after precipitation the casein) by affinity purification. To evaluate the effectiveness of the p-aminophenol modified magnetic nanocarriers, the three different types of nanocarriers with different linkers having varying lengths were prepared. All nanocarriers were characterized and validated using FT-IR, SEM, EDX, VSM and XRD analysis methods According to our results, p-aminophenol ligand attached to the nanocarrier by long linkers better separates the α-amylase from the casein free skim milk with 49.66% recovery and 48.18-fold purification efficiency. The results of this study showed that our novel magnetic nanocarriers have the capacity to be used for fast, reproducible and cost-effective purification of α-amylase.
目前，改性FeO磁性纳米颗粒经常被用作蛋白质和酶纯化的纳米载体。在本研究中，制备了FeO磁性纳米颗粒，并用通过不同连接子固定的对氨基苯酚亲和配体对其表面进行了改性。改性后的纳米载体用于通过亲和纯化从牛奶（酪蛋白沉淀后）中纯化α-淀粉酶。为了评估对氨基苯酚改性磁性纳米载体的有效性，制备了三种具有不同长度连接子的不同类型的纳米载体。使用FT-IR、SEM、EDX、VSM和XRD分析方法对所有纳米载体进行了表征和验证。根据我们的结果，通过长连接子连接到纳米载体上的对氨基苯酚配体能更好地从无酪蛋白的脱脂牛奶中分离α-淀粉酶，回收率为49.66%，纯化效率为48.18倍。本研究结果表明，我们的新型磁性纳米载体有能力用于快速、可重复且经济高效地纯化α-淀粉酶。
