Çelik Safinur Yıldırım, Solak Kübra, Mavi Ahmet
Department of Nutrition and Dietetics, Faculty of Health Sciences, Atatürk University, Erzurum, Turkey.
Department of Nanoscience and Nanoengineering, Graduate School of Natural and Applied Sciences, Atatürk University, Erzurum, Turkey.
Appl Biochem Biotechnol. 2023 Jun;195(6):3900-3913. doi: 10.1007/s12010-022-03983-9. Epub 2022 Jun 1.
Magnetic nanoparticles (MNPs) have been used for purification of specific biomolecules form mixtures. The aim of this study is to develop a new, cheap, reusable, and magnetic-based material to purify the carbonic anhydrase (CA) enzyme in a short time with high efficiency. In the first part of this study, silica-coated iron oxide magnetic nanoparticles (FeO@SiO MNPs) were obtained. Surface modification of FeO@SiO MNPs was accomplished with 3-(4-Hydroxyphenyl) propionic acid (PA) and sulfanilamide (SA), respectively. SA is a selective inhibitor of CA, and it selectively binds to CA. The final particle was named FeO@SiO-PA-SA MNPs and characterized by SEM, TEM, XRD, and FT-IR. It was determined that the produced MNPs contained multicore, were smaller than 100 nm in size, and had a spherical morphology. The CA was purified from bovine blood hemolysate in a short time such as 2.5 h and in a simple manner. The maximum enzyme purifying capacity of MNPs was calculated as 13.87 ± 3.27 mg CA/g MNP. SDS-PAGE analysis was confirmed that high CA purification success was achieved.
磁性纳米颗粒(MNPs)已被用于从混合物中纯化特定生物分子。本研究的目的是开发一种新型、廉价、可重复使用的磁性材料,以便在短时间内高效纯化碳酸酐酶(CA)。在本研究的第一部分,制备了二氧化硅包覆的氧化铁磁性纳米颗粒(FeO@SiO MNPs)。分别用3-(4-羟基苯基)丙酸(PA)和磺胺(SA)对FeO@SiO MNPs进行表面修饰。SA是CA的选择性抑制剂,它能选择性地与CA结合。最终的颗粒被命名为FeO@SiO-PA-SA MNPs,并通过扫描电子显微镜(SEM)、透射电子显微镜(TEM)、X射线衍射(XRD)和傅里叶变换红外光谱(FT-IR)进行表征。结果表明,所制备的MNPs为多核结构,尺寸小于100 nm,呈球形。CA能在2.5小时等短时间内以简单的方式从牛血溶血产物中纯化出来。MNPs的最大酶纯化能力经计算为13.87±3.27 mg CA/g MNP。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析证实,CA的纯化取得了很高的成功率。